This document summarizes a research article that investigated whether disrupting the circadian rhythms of rats by exposing them to a 22-hour light-dark cycle leads to depressive-like behaviors. The study found that rats exposed to the disrupted light cycle showed signs of depression, including decreased pleasure-seeking behaviors, sexual dysfunction, and increased immobility in tests for depressive symptoms. Analysis also revealed changes in brain chemistry related to mood. The results support the hypothesis that misalignment of circadian rhythms due to problems with the brain's central clock may contribute to the development of depression.
The Implications of Desynchronized Circadian Rhythms in Human Mental Health a...Cristina Corlito
The organization of the mammalian circadian system relies on temporal order between behavioural and physiological rhythms that are critical to the normal functioning of the body and human health. The hypothesis proposed is that a disruption in the sleep-wake cycle reflects impaired circadian clock functioning, which synergistically leads to the progression and maintenance of a variety of psychiatric disorders. The aging population is most susceptible to the depletion of chronobiological rhythms and sleep deficits, and thus, the development of psychiatric disorders in the elderly warrants attention. By evaluating previous and current literature, it was found that internal temporal disorder in humans may result from both internal and external factors that disrupt the coordinated symphony of the SCN and peripheral oscillators. Sleep disorders and neuropsychiatric illnesses transpire as a result of this chronodisruption. Evidence suggests that sleep disturbances are a causal factor of psychiatric illness, rather than being mere complications. It is proposed that senescence not only predisposes the elderly to chronodisruption and sleep deficits, but also increases their risk for developing frequently comorbid psychiatric illnesses. Increasing public awareness of the multitude of strategies available for harmonious synchronization and optimal well-being are profitable to the elderly in preventing circadian malfunction.
The influence of the HPA axis in development of psychosisAshley Brodell
This document discusses the influence of the hypothalamic-pituitary-adrenal (HPA) axis on the development of psychosis. It notes that psychosis sufferers often have elevated HPA axis activity and cortisol levels, especially during stressful periods and psychotic episodes. Long-term HPA axis dysregulation can lead to changes in the brain like reduced hippocampal volume and increased dopamine levels, contributing to psychosis symptoms. While cortisol levels may decrease in chronic cases, the HPA axis still plays a key role in the diathesis stress model of psychosis by mediating the effects of genetics and environment on mental illness onset and progression.
This document summarizes Storeganizer, an innovative storage system that provides high-density storage in warehouse racks. Storeganizer allows for 40% more usable storage space by stacking items in vertical columns up to three rows deep. It provides optimal picking and less lifting/bending. Storeganizer is a universal system that can be easily installed in any existing rack. It offers tailored sizes and specifications for different inventory needs.
This document discusses circadian rhythm dysregulation in bipolar disorder. It summarizes that circadian rhythms control physiological and behavioral cycles over 24 hours, and disturbances in these rhythms can contribute to psychiatric disorders like bipolar disorder. Evidence from animal and clinical studies supports a link between circadian abnormalities and bipolar disorder. Specifically, patients with bipolar disorder may experience a shortening of the circadian period length which can lead to a desynchronization of internal rhythms with the external light-dark cycle. This misalignment has been correlated with the switching between manic and depressive mood states in bipolar disorder.
Circadian rhythm refers to biological cycles that occur over approximately 24 hours. These rhythms are regulated by molecular feedback loops in clock genes and proteins that influence cellular functions and synchronize organs. Disruption of circadian rhythms through irregular sleep/wake cycles, jet lag, or light exposure at night has been linked to increased risk of metabolic, cardiovascular, and mental health conditions. Maintaining circadian alignment through regular sleep/wake and meal times may help reduce disease risk.
The Role Of Cytokines On Immune PrivilegeKaty Allen
- Immune privilege sites like the brain actively suppress inflammation to protect delicate tissues from damage. Cytokines play a role in maintaining this immune privilege.
- Experiments on mice found that chronic early-life stress impaired microglial function and rewired the brain's stress response pathways, causing depression-like behaviors in adulthood. Treating the stress hormone CRH reversed these effects.
- Exposure to toxins in tobacco smoke during development can alter brain cell proliferation, synaptic activity, and microglial function, potentially leading to neurological and cognitive impairments.
Appetitive vs Aversive Motivation in AddictionPaul Coelho, MD
This document discusses addiction from a neurobiological and motivational perspective. It proposes that addiction involves a cycle of decreased function of brain reward systems and recruitment of antireward systems that progressively worsen. Excessive drug use results in short-term relief of the reward deficit but long-term worsening of neurochemical dysregulations, forming an allostatic state reflected in a chronic deviation of reward set point fueled by dysregulation of reward circuits and recruitment of stress responses. Vulnerability to addiction may involve genetic and developmental factors that sensitize brain antireward systems.
Molecular mechanisms that control circadian rhythms - Mohammed Elreishi Mohammed Elreishi
Circadian rhythms are driven by an internal
biological clock that anticipates day/night cycles to
optimize the physiology and behavior of organisms.
The 2017 Nobel Prize in Physiology or Medicine is
awarded to Jeffrey C. Hall, Michael Rosbash and
Michael W. Young for their Discoveries of Molecular Mechanisms Controlling the Circadian Rhythm.
The Implications of Desynchronized Circadian Rhythms in Human Mental Health a...Cristina Corlito
The organization of the mammalian circadian system relies on temporal order between behavioural and physiological rhythms that are critical to the normal functioning of the body and human health. The hypothesis proposed is that a disruption in the sleep-wake cycle reflects impaired circadian clock functioning, which synergistically leads to the progression and maintenance of a variety of psychiatric disorders. The aging population is most susceptible to the depletion of chronobiological rhythms and sleep deficits, and thus, the development of psychiatric disorders in the elderly warrants attention. By evaluating previous and current literature, it was found that internal temporal disorder in humans may result from both internal and external factors that disrupt the coordinated symphony of the SCN and peripheral oscillators. Sleep disorders and neuropsychiatric illnesses transpire as a result of this chronodisruption. Evidence suggests that sleep disturbances are a causal factor of psychiatric illness, rather than being mere complications. It is proposed that senescence not only predisposes the elderly to chronodisruption and sleep deficits, but also increases their risk for developing frequently comorbid psychiatric illnesses. Increasing public awareness of the multitude of strategies available for harmonious synchronization and optimal well-being are profitable to the elderly in preventing circadian malfunction.
The influence of the HPA axis in development of psychosisAshley Brodell
This document discusses the influence of the hypothalamic-pituitary-adrenal (HPA) axis on the development of psychosis. It notes that psychosis sufferers often have elevated HPA axis activity and cortisol levels, especially during stressful periods and psychotic episodes. Long-term HPA axis dysregulation can lead to changes in the brain like reduced hippocampal volume and increased dopamine levels, contributing to psychosis symptoms. While cortisol levels may decrease in chronic cases, the HPA axis still plays a key role in the diathesis stress model of psychosis by mediating the effects of genetics and environment on mental illness onset and progression.
This document summarizes Storeganizer, an innovative storage system that provides high-density storage in warehouse racks. Storeganizer allows for 40% more usable storage space by stacking items in vertical columns up to three rows deep. It provides optimal picking and less lifting/bending. Storeganizer is a universal system that can be easily installed in any existing rack. It offers tailored sizes and specifications for different inventory needs.
This document discusses circadian rhythm dysregulation in bipolar disorder. It summarizes that circadian rhythms control physiological and behavioral cycles over 24 hours, and disturbances in these rhythms can contribute to psychiatric disorders like bipolar disorder. Evidence from animal and clinical studies supports a link between circadian abnormalities and bipolar disorder. Specifically, patients with bipolar disorder may experience a shortening of the circadian period length which can lead to a desynchronization of internal rhythms with the external light-dark cycle. This misalignment has been correlated with the switching between manic and depressive mood states in bipolar disorder.
Circadian rhythm refers to biological cycles that occur over approximately 24 hours. These rhythms are regulated by molecular feedback loops in clock genes and proteins that influence cellular functions and synchronize organs. Disruption of circadian rhythms through irregular sleep/wake cycles, jet lag, or light exposure at night has been linked to increased risk of metabolic, cardiovascular, and mental health conditions. Maintaining circadian alignment through regular sleep/wake and meal times may help reduce disease risk.
The Role Of Cytokines On Immune PrivilegeKaty Allen
- Immune privilege sites like the brain actively suppress inflammation to protect delicate tissues from damage. Cytokines play a role in maintaining this immune privilege.
- Experiments on mice found that chronic early-life stress impaired microglial function and rewired the brain's stress response pathways, causing depression-like behaviors in adulthood. Treating the stress hormone CRH reversed these effects.
- Exposure to toxins in tobacco smoke during development can alter brain cell proliferation, synaptic activity, and microglial function, potentially leading to neurological and cognitive impairments.
Appetitive vs Aversive Motivation in AddictionPaul Coelho, MD
This document discusses addiction from a neurobiological and motivational perspective. It proposes that addiction involves a cycle of decreased function of brain reward systems and recruitment of antireward systems that progressively worsen. Excessive drug use results in short-term relief of the reward deficit but long-term worsening of neurochemical dysregulations, forming an allostatic state reflected in a chronic deviation of reward set point fueled by dysregulation of reward circuits and recruitment of stress responses. Vulnerability to addiction may involve genetic and developmental factors that sensitize brain antireward systems.
Molecular mechanisms that control circadian rhythms - Mohammed Elreishi Mohammed Elreishi
Circadian rhythms are driven by an internal
biological clock that anticipates day/night cycles to
optimize the physiology and behavior of organisms.
The 2017 Nobel Prize in Physiology or Medicine is
awarded to Jeffrey C. Hall, Michael Rosbash and
Michael W. Young for their Discoveries of Molecular Mechanisms Controlling the Circadian Rhythm.
1) The document discusses circadian rhythms, which are biological processes that display 24-hour oscillations driven by an internal biological clock.
2) It reviews how the suprachiasmatic nucleus in the hypothalamus controls circadian rhythms in mammals and can be entrained by external factors like light and temperature.
3) Disruptions to circadian rhythms can cause health issues like diabetes, obesity, and psychological disorders. Certain drugs like cocaine can also disrupt circadian rhythms.
This summary analyzes a scientific article that uses proteomics analysis to study molecular and cellular mechanisms associated with anxiety in the cingulate cortex synapses of mice with high versus low anxiety-related behaviors. The key findings are:
1) Proteins overrepresented in high anxiety mice indicate stabilization and enlargement of existing excitatory dendritic spines through reinforcement of glutamatergic synaptic structures and long-term potentiation maintenance mechanisms.
2) There is evidence of increased spontaneous synaptic activity in non-glutamatergic networks coupled with relatively weakened GABAergic inhibitory control.
3) Mitochondrial proteins point to increased oxidative phosphorylation, fatty acid oxidation, and ATP production, suggesting adaptations to persistent synaptic
The document discusses the function of memory and how stress and noradrenergic mechanisms affect memory formation and consolidation. It states that the stress response is mediated by the HPA axis and release of catecholamines, which activate pathways that stimulate long-term memory formation. However, exposure to traumatic or chronic stress can impair memory or cause memory-related disorders like PTSD. The noradrenergic system is implicated in emotional memory consolidation. Administering beta-adrenergic receptor antagonists during memory reconsolidation is a possible method for modifying intolerable memories in psychiatric patients. The paper will characterize the effects of the beta-blocker propranolol on stress-mediated memory reconsolidation.
Circadian Rhythm Enhancing Brain Synchronization and Cognitionijtsrd
Cognition is impaired in many neuropsychiatric disorders and the quality of life is severely affected. A key mechanism for sculpting communication patterns between large scale brain networks that underpin cognition and its breakdown in neuropsychiatric disorders is synchronous electrophysiological rhythms. According to a study, light has a wide range of effects on the synchronization of circadian rhythms with the external environment and it is found that light influences the urinary excretion of melatonin and controls sleep. Autonomic and neuroendocrine responses such as feedback regulation and the involvement of the immune system have also been shown to influence the circadian rhythm. There have been major advances in our understanding of the retinal photoreceptors mediating these non image forming light responses over the last two decades, as well as the neural pathways and molecular mechanisms that generate and energize circadian rhythms in the phase of the light dark LD cycle. Our understanding of the mechanisms by which lighting impacts cognitive processes, on the other hand, is more misleading. Lights effect on different cognitive processes is complex. Indirect effects may also arise due to disrupted circadian rhythm, in addition to the direct effects of light on consciousness. In studies that rely on various cognitive and sensory processes, different assays have been used, which can also contribute to variable outcomes. The physiological basis of these responses and the influence of various lighting environments on cognitive processes are summarized here, taking into account their effects on circadian rhythms, sleep and arousal. Uthirakumar Devaraj | Elumalai Balamurugan "Circadian Rhythm: Enhancing Brain Synchronization and Cognition" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-6 , October 2021, URL: https://www.ijtsrd.com/papers/ijtsrd46394.pdf Paper URL : https://www.ijtsrd.com/biological-science/neurobiology/46394/circadian-rhythm-enhancing-brain-synchronization-and-cognition/uthirakumar-devaraj
This study seeks to analyze the molecular basis of circadian rhythms and behavioral modulation by hormones in the nudibranch Melibe leonina. The goals are to localize circadian clock neurons in the central nervous system using immunohistochemistry, stain for the neurotransmitters conopressin and melatonin, and assess behavioral responses to injections of these hormones through video recordings. Understanding the relationship between clock neurons, hormones, and behavior can provide insights into circadian rhythm regulation applicable to fields like medicine.
Cytokine purine interactions in behavioral depression in ratszpzp0312
This paper reviews recent research on metabolic and immune mediators of behavioral depression in rats induced by reserpine injection. The paper finds that reserpine injection causes behavioral depression in rats, as evidenced by increased time spent floating in a swim test. This depression occurs in two components - an early component 1 hour after injection, and a later component 48 hours after injection. The early component is not reversed by blocking the interleukin-1β receptor, but the later component is reversed, suggesting it is mediated by the proinflammatory cytokine interleukin-1β. Both components are reversed by blocking the adenosine A2A receptor, suggesting interactions between purine and cytokine signaling pathways in behavioral depression.
Circadian rhythm refers to the approximately 24-hour cycles in human and animal physiology and behavior, regulated by an internal biological clock. The master circadian clock is located in the hypothalamus, specifically the suprachiasmatic nucleus, which receives light input from the retina. These circadian rhythms evolved to protect DNA from UV radiation and help entrain organisms to the light-dark cycle. Core body temperature, melatonin secretion, and cortisol levels are classic markers used to measure circadian rhythms. Light exposure can advance or delay circadian rhythms depending on timing.
Circadian rhythms exhibit a period of about 24 h whereas circannual (seasonal) rhythms cycle over the course of a year. Both types of rhythms are coordinated by the brain's suprachiasmatic nucleus (SCN).
This document provides an overview of measuring autonomic nervous system regulation through various physiological tools. It begins with background on the autonomic nervous system and its role in the stress response. Tools that can measure aspects of the autonomic nervous system like cardiac, skin conductance, respiratory, and vascular activity are identified. A decision tree is provided to help choose the best tools depending on setting, purpose, ease of use, and validity. The goal is to use these tools to measure changes in autonomic nervous system activation from techniques like mind-body skills and monitor ability to return to homeostasis after stress.
The reticular activating system (RAS) is a complex network of neurons located in the brainstem that regulates states of consciousness such as sleep and wakefulness. It is composed of neurotransmitter systems including cholinergic, adrenergic, serotonergic, and histaminergic neurons. The RAS maintains arousal and attention by transmitting signals from the brainstem to the thalamus and cortex. Disruptions in the RAS can result in disturbances of consciousness and sleep-wake cycles, and have been implicated in disorders such as schizophrenia, PTSD, Parkinson's disease, and Alzheimer's disease.
- GnRH neuronal cell bodies are located in the hypothalamus and extend processes to the median eminence where GnRH is released into the hypophyseal portal system.
- The hypophyseal portal system connects hypothalamic GnRH neurons to gonadotropes in the anterior pituitary.
- GnRH stimulates the anterior pituitary to secrete LH and FSH, which then direct gonadal function in the ovaries and testes.
Physiology of Neuromodulation and neuromodulators. Difference between neuromodulation and synapse. Recent advances in neuromodulation, clinical application of neuromodulation.
This document discusses a proposed theory using Turing machines to develop a new therapeutic strategy for treating some spinal cord and brain conditions using a depurative-toxicological-pharmacological approach. It reviews literature on neurodegenerative diseases like Alzheimer's, Parkinson's, and ALS to understand common pathological mechanisms involving accumulation of toxic metabolic byproducts that the central nervous system lacks efficient means of removing. The theory aims to translate these insights into a practical hypothesis for reducing or delaying disease progression. The document also summarizes findings from studies showing cerebrospinal fluid from progressive multiple sclerosis patients causes mitochondrial dysfunction in neurons, identifying a potential biological mechanism and therapeutic target for progressive forms of the disease.
Etiology and neurobiology of obsessive compulsive disorderSimranSandhu673667
This document provides an overview of the etiology and neurobiology of obsessive-compulsive disorder (OCD). It discusses the key topics of diagnosis, neurobiological theories, psychodynamic theories, learning-based theories, cognitive theories, and summary. The neurobiology section outlines the neuroanatomy of OCD including brain regions like the anterior cingulate cortex, orbitofrontal cortex, basal ganglia, and cortico-striatal-thalamic circuits. It also discusses the neurochemistry involving serotonin, dopamine, and glutamate. Neuroimaging findings from MRI, fMRI, and DTI studies are summarized as well. Finally, psychodynamic theories involving Freud's psychosexual stages of development and the case
This document discusses using audio-visual entrainment (AVE) to treat seasonal affective disorder (SAD). It provides background on SAD, noting it affects 6-20% of northern populations and has similar symptoms in other regions/contexts. Traditional treatments are light therapy and antidepressants. The document outlines a study applying 20Hz AVE to 74 SAD patients over 4 weeks, finding it reduced depression and anxiety symptoms more than a placebo session. AVE is proposed as an effective alternative or addition to light therapy for treating SAD.
The document summarizes a study that investigated whether neurodegeneration alone can cause schizophrenia-like behaviors in mice. The study ablated neural stem cells in transgenic mice using ganciclovir over 56 days. Behavioral tests revealed no significant differences between control and ablated mice in prepulse inhibition, locomotor activity after PCP/amphetamine, time spent in open arms of elevated plus maze, or floating behavior in Porsolt swim test. This suggests neurodegeneration alone may not cause schizophrenia. However, other studies that also applied stressors found ablation did produce schizophrenia-like behaviors, so stress may be a contributing factor.
The document discusses an Indigenous model of traditional wellness called the Medicine Wheel and how it relates to neurodecolonization. It covers how colonialism has impacted Indigenous peoples' brains, bodies, and genes through trauma, and how engaging in traditional mindfulness practices can help heal from these impacts by restructuring and decolonizing the mind and body. Some key traditional practices discussed that can facilitate this include running, dancing, singing, sleeping, laughing, fasting, meditation, time in nature, and spending time in community.
This document is an abstract submission for a speech on common aspects in the physiological pathways of addiction and evolutionary processes that are useful for pharmaco-toxicological approaches. It discusses how certain neuronal circuits play a crucial role in addiction, learning, and reward mechanisms in mammals, as seen in behaviors like breastfeeding and parental care of offspring. Several studies are cited showing the involvement of dopamine and other neurotransmitters in addiction and natural reward processes, suggesting similarities that can help understand and potentially treat addiction.
1) The document discusses circadian rhythms, which are biological processes that display 24-hour oscillations driven by an internal biological clock.
2) It reviews how the suprachiasmatic nucleus in the hypothalamus controls circadian rhythms in mammals and can be entrained by external factors like light and temperature.
3) Disruptions to circadian rhythms can cause health issues like diabetes, obesity, and psychological disorders. Certain drugs like cocaine can also disrupt circadian rhythms.
This summary analyzes a scientific article that uses proteomics analysis to study molecular and cellular mechanisms associated with anxiety in the cingulate cortex synapses of mice with high versus low anxiety-related behaviors. The key findings are:
1) Proteins overrepresented in high anxiety mice indicate stabilization and enlargement of existing excitatory dendritic spines through reinforcement of glutamatergic synaptic structures and long-term potentiation maintenance mechanisms.
2) There is evidence of increased spontaneous synaptic activity in non-glutamatergic networks coupled with relatively weakened GABAergic inhibitory control.
3) Mitochondrial proteins point to increased oxidative phosphorylation, fatty acid oxidation, and ATP production, suggesting adaptations to persistent synaptic
The document discusses the function of memory and how stress and noradrenergic mechanisms affect memory formation and consolidation. It states that the stress response is mediated by the HPA axis and release of catecholamines, which activate pathways that stimulate long-term memory formation. However, exposure to traumatic or chronic stress can impair memory or cause memory-related disorders like PTSD. The noradrenergic system is implicated in emotional memory consolidation. Administering beta-adrenergic receptor antagonists during memory reconsolidation is a possible method for modifying intolerable memories in psychiatric patients. The paper will characterize the effects of the beta-blocker propranolol on stress-mediated memory reconsolidation.
Circadian Rhythm Enhancing Brain Synchronization and Cognitionijtsrd
Cognition is impaired in many neuropsychiatric disorders and the quality of life is severely affected. A key mechanism for sculpting communication patterns between large scale brain networks that underpin cognition and its breakdown in neuropsychiatric disorders is synchronous electrophysiological rhythms. According to a study, light has a wide range of effects on the synchronization of circadian rhythms with the external environment and it is found that light influences the urinary excretion of melatonin and controls sleep. Autonomic and neuroendocrine responses such as feedback regulation and the involvement of the immune system have also been shown to influence the circadian rhythm. There have been major advances in our understanding of the retinal photoreceptors mediating these non image forming light responses over the last two decades, as well as the neural pathways and molecular mechanisms that generate and energize circadian rhythms in the phase of the light dark LD cycle. Our understanding of the mechanisms by which lighting impacts cognitive processes, on the other hand, is more misleading. Lights effect on different cognitive processes is complex. Indirect effects may also arise due to disrupted circadian rhythm, in addition to the direct effects of light on consciousness. In studies that rely on various cognitive and sensory processes, different assays have been used, which can also contribute to variable outcomes. The physiological basis of these responses and the influence of various lighting environments on cognitive processes are summarized here, taking into account their effects on circadian rhythms, sleep and arousal. Uthirakumar Devaraj | Elumalai Balamurugan "Circadian Rhythm: Enhancing Brain Synchronization and Cognition" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-6 , October 2021, URL: https://www.ijtsrd.com/papers/ijtsrd46394.pdf Paper URL : https://www.ijtsrd.com/biological-science/neurobiology/46394/circadian-rhythm-enhancing-brain-synchronization-and-cognition/uthirakumar-devaraj
This study seeks to analyze the molecular basis of circadian rhythms and behavioral modulation by hormones in the nudibranch Melibe leonina. The goals are to localize circadian clock neurons in the central nervous system using immunohistochemistry, stain for the neurotransmitters conopressin and melatonin, and assess behavioral responses to injections of these hormones through video recordings. Understanding the relationship between clock neurons, hormones, and behavior can provide insights into circadian rhythm regulation applicable to fields like medicine.
Cytokine purine interactions in behavioral depression in ratszpzp0312
This paper reviews recent research on metabolic and immune mediators of behavioral depression in rats induced by reserpine injection. The paper finds that reserpine injection causes behavioral depression in rats, as evidenced by increased time spent floating in a swim test. This depression occurs in two components - an early component 1 hour after injection, and a later component 48 hours after injection. The early component is not reversed by blocking the interleukin-1β receptor, but the later component is reversed, suggesting it is mediated by the proinflammatory cytokine interleukin-1β. Both components are reversed by blocking the adenosine A2A receptor, suggesting interactions between purine and cytokine signaling pathways in behavioral depression.
Circadian rhythm refers to the approximately 24-hour cycles in human and animal physiology and behavior, regulated by an internal biological clock. The master circadian clock is located in the hypothalamus, specifically the suprachiasmatic nucleus, which receives light input from the retina. These circadian rhythms evolved to protect DNA from UV radiation and help entrain organisms to the light-dark cycle. Core body temperature, melatonin secretion, and cortisol levels are classic markers used to measure circadian rhythms. Light exposure can advance or delay circadian rhythms depending on timing.
Circadian rhythms exhibit a period of about 24 h whereas circannual (seasonal) rhythms cycle over the course of a year. Both types of rhythms are coordinated by the brain's suprachiasmatic nucleus (SCN).
This document provides an overview of measuring autonomic nervous system regulation through various physiological tools. It begins with background on the autonomic nervous system and its role in the stress response. Tools that can measure aspects of the autonomic nervous system like cardiac, skin conductance, respiratory, and vascular activity are identified. A decision tree is provided to help choose the best tools depending on setting, purpose, ease of use, and validity. The goal is to use these tools to measure changes in autonomic nervous system activation from techniques like mind-body skills and monitor ability to return to homeostasis after stress.
The reticular activating system (RAS) is a complex network of neurons located in the brainstem that regulates states of consciousness such as sleep and wakefulness. It is composed of neurotransmitter systems including cholinergic, adrenergic, serotonergic, and histaminergic neurons. The RAS maintains arousal and attention by transmitting signals from the brainstem to the thalamus and cortex. Disruptions in the RAS can result in disturbances of consciousness and sleep-wake cycles, and have been implicated in disorders such as schizophrenia, PTSD, Parkinson's disease, and Alzheimer's disease.
- GnRH neuronal cell bodies are located in the hypothalamus and extend processes to the median eminence where GnRH is released into the hypophyseal portal system.
- The hypophyseal portal system connects hypothalamic GnRH neurons to gonadotropes in the anterior pituitary.
- GnRH stimulates the anterior pituitary to secrete LH and FSH, which then direct gonadal function in the ovaries and testes.
Physiology of Neuromodulation and neuromodulators. Difference between neuromodulation and synapse. Recent advances in neuromodulation, clinical application of neuromodulation.
This document discusses a proposed theory using Turing machines to develop a new therapeutic strategy for treating some spinal cord and brain conditions using a depurative-toxicological-pharmacological approach. It reviews literature on neurodegenerative diseases like Alzheimer's, Parkinson's, and ALS to understand common pathological mechanisms involving accumulation of toxic metabolic byproducts that the central nervous system lacks efficient means of removing. The theory aims to translate these insights into a practical hypothesis for reducing or delaying disease progression. The document also summarizes findings from studies showing cerebrospinal fluid from progressive multiple sclerosis patients causes mitochondrial dysfunction in neurons, identifying a potential biological mechanism and therapeutic target for progressive forms of the disease.
Etiology and neurobiology of obsessive compulsive disorderSimranSandhu673667
This document provides an overview of the etiology and neurobiology of obsessive-compulsive disorder (OCD). It discusses the key topics of diagnosis, neurobiological theories, psychodynamic theories, learning-based theories, cognitive theories, and summary. The neurobiology section outlines the neuroanatomy of OCD including brain regions like the anterior cingulate cortex, orbitofrontal cortex, basal ganglia, and cortico-striatal-thalamic circuits. It also discusses the neurochemistry involving serotonin, dopamine, and glutamate. Neuroimaging findings from MRI, fMRI, and DTI studies are summarized as well. Finally, psychodynamic theories involving Freud's psychosexual stages of development and the case
This document discusses using audio-visual entrainment (AVE) to treat seasonal affective disorder (SAD). It provides background on SAD, noting it affects 6-20% of northern populations and has similar symptoms in other regions/contexts. Traditional treatments are light therapy and antidepressants. The document outlines a study applying 20Hz AVE to 74 SAD patients over 4 weeks, finding it reduced depression and anxiety symptoms more than a placebo session. AVE is proposed as an effective alternative or addition to light therapy for treating SAD.
The document summarizes a study that investigated whether neurodegeneration alone can cause schizophrenia-like behaviors in mice. The study ablated neural stem cells in transgenic mice using ganciclovir over 56 days. Behavioral tests revealed no significant differences between control and ablated mice in prepulse inhibition, locomotor activity after PCP/amphetamine, time spent in open arms of elevated plus maze, or floating behavior in Porsolt swim test. This suggests neurodegeneration alone may not cause schizophrenia. However, other studies that also applied stressors found ablation did produce schizophrenia-like behaviors, so stress may be a contributing factor.
The document discusses an Indigenous model of traditional wellness called the Medicine Wheel and how it relates to neurodecolonization. It covers how colonialism has impacted Indigenous peoples' brains, bodies, and genes through trauma, and how engaging in traditional mindfulness practices can help heal from these impacts by restructuring and decolonizing the mind and body. Some key traditional practices discussed that can facilitate this include running, dancing, singing, sleeping, laughing, fasting, meditation, time in nature, and spending time in community.
This document is an abstract submission for a speech on common aspects in the physiological pathways of addiction and evolutionary processes that are useful for pharmaco-toxicological approaches. It discusses how certain neuronal circuits play a crucial role in addiction, learning, and reward mechanisms in mammals, as seen in behaviors like breastfeeding and parental care of offspring. Several studies are cited showing the involvement of dopamine and other neurotransmitters in addiction and natural reward processes, suggesting similarities that can help understand and potentially treat addiction.
2. 1
CIRCADIAN FORCED DESYNCHRONY OF THE MASTER CLOCK LEADS1
TO PHENOTYPIC MANIFESTATION OF DEPRESSION IN RATS2
Abbreviated title: Circadian desynchrony leads to depression3
Miriam Ben-Hamo1,2
, Tracy Larson1
, Leanne S. Duge1
, Carl Sikkema3
, Charles W.4
Wilkinson2,3
, Horacio O. de la Iglesia1,2
and Mónica M.C. González1,4
5
1
Department of Biology and Program in Neurobiology and Behavior, University of6
Washington, Seattle, WA 981957
2
Program in Neurobiology and Behavior, University of Washington, Seattle, WA 981958
3
Geriatric Research, Education and Clinical Center, VA Puget Sound Health Care9
System, Seattle, WA 9819510
4
Current Address: Instituto Ferrero de Neurología y Sueño, Buenos Aires, Argentina11
*Correspondence to: H.O. de la Iglesia (horaciod@uw.edu) and M.M.C. González12
(mgonzalez@ifn.com.ar)13
14
Author contributions: MB-H, HOD and MMCG designed research, performed research,15
analyzed data and wrote the paper. TL and CWW performed research and wrote paper.16
LSD and CS performed research.17
18
3. 2
ABSTRACT19
In mammals, a master circadian clock within the suprachiasmatic nucleus (SCN) of20
the hypothalamus maintains the phase coherence among a wide array of behavioral and21
physiological circadian rhythms. Affective disorders are typically associated with22
disruption of this fine-tuned ‘internal synchronization’, but whether this internal23
misalignment is part of the physiopathology of mood disorders is not clear. To date,24
depressive-like behavior in animal models has been induced by methods that fail to25
specifically target the SCN regulation of internal synchronization as the mode to generate26
depression. In the rat, exposure to a 22h light-dark cycle (LD22) leads to the uncoupling27
of two distinct populations of neuronal oscillators within the SCN. This genetically,28
neurally and pharmacologically intact animal model represents a unique opportunity to29
assess the effect of a systematic challenge to the central circadian pacemaker on30
phenotypic manifestations of mood disorders. We show that LD22 circadian forced31
desynchrony in rats induces depressive-like phenotypes including anhedonia, sexual32
dysfunction, and increased immobility in the forced swim test (FST), as well as changes33
in the levels and turnover rates of monoamines within the prefrontal cortex.34
Desynchronized rats show increased FST immobility during the dark (active) phase but35
decreased immobility during the light (rest) phase, suggesting a decrease in the amplitude36
of the normal daily oscillation in this behavioral manifestation of depression. Our results37
support the notion that the prolonged internal misalignment of circadian rhythms induced38
by environmental challenge to the central circadian pacemaker may constitute part of the39
etiology of depression.40
4. 3
SIGNIFICANCE STATEMENT41
Previous studies hypothesized that disruption of the internal coordination of circadian42
rhythms by the master circadian clock may contribute to the etiology of mood disorders.43
This hypothesis has not yet been tested in genetically and neurologically intact animal44
models with impairment of the master clock’s ability to maintain circadian internal45
synchronization. We compared rats exposed to 22h light-dark (LD) cycle, which leads to46
forced desynchrony of neuronal oscillators within the suprachiasmatic nucleus, to rats47
exposed to a normal 24h LD cycle. We show that forced desynchrony induces a48
depressive phenotype that is particularly manifested during the dark (active) phase of the49
animals. Our results demonstrate that prolonged internal misalignment of circadian50
rhythms may constitute part of the etiology of depression.51
5. 4
INTRODUCTION52
In mammals, circadian rhythms are coordinated by a master circadian clock located in53
the suprachiasmatic nucleus (SCN), where transcriptional and translational feedback54
loops within single cells oscillate with a period of approximately 24 h; these cells55
compose a neuronal network that generates robust endogenous circadian oscillations56
(Colwell, 2011). Direct retinal projections to the SCN via the retinohypothalamic tract57
(RHT) assure entrainment of the master clock—as well as the body rhythms it58
regulates—to the light-dark (LD) cycle (Golombek and Rosenstein, 2010). In this59
manner, the SCN governs a wide array of behavioral and physiological rhythms including60
the sleep-wake cycle, body temperature, hormone release, appetite and cognitive61
functions (Saper et al., 2005). This regulation is achieved directly or through inputs to62
both central and peripheral extra-SCN circadian oscillators that locally time tissue-63
specific processes (Mohawk et al., 2012). Thus, the SCN master clock is responsible for64
maintaining phase coherence within the complex network of body oscillators and the65
rhythms they regulate. Disruption of this fine-tuned ‘internal synchronization’ is often66
associated with both somatic and neuropsychiatric disorders (Takahashi et al., 2008;67
Wulff et al., 2010; McCarthy and Welsh, 2012; Morris et al., 2012; McClung, 2013).68
Numerous lines of evidence support the association between abnormal endogenous69
circadian rhythms and affective disorders. Patients with affective disorders show70
disruptions in an array of circadian rhythms including sleep, body temperature rhythm,71
and hormone release (Boivin, 2000; Wirz-Justice, 2006), suggesting that mood disorders72
are linked to impairment of the master regulation of circadian rhythms rather than the73
disruption of isolated rhythmic outputs. Moreover, bright light therapy, sleep deprivation,74
6. 5
sleep phase advance and social rhythm therapy, which are all capable of resetting the75
phase of the master circadian clock, were shown to be effective in the treatment of76
affective disorders (Riemann et al., 2002; Frank et al., 2005; Golden et al., 2005;77
Hemmeter et al., 2010). In addition, the SCN projects directly or through relays to the78
prefrontal cortex, hippocampus and amygdala (Leak and Moore, 2001), and trans-79
synaptically drives the circadian cellular activity of monoaminergic brain regions (Aston-80
Jones et al., 2001; Luo and Aston‐Jones, 2009), all of which are associated with mood81
regulation. Accordingly, exposure to short photoperiods or constant light can lead to82
behavioral symptoms of depression in different rodent models (Prendergast and Nelson,83
2005; McClung, 2011), and rats exposed to constant darkness for several weeks show84
apoptosis in mood-related monoaminergic neurons in association to a depressive85
behavioral phenotype (Gonzalez and Aston-Jones, 2006, 2008). These environmental86
light effects were attributed in part to the master circadian clock. Finally, mice with87
mutated or knocked down clock genes typically show abnormal affective behavior88
(Prickaerts et al., 2006; Roybal et al., 2007; Keers et al., 2012). These behavioral effects89
emerge, at least in part, from the absence of a functional clock within SCN neurons, as90
region-specific knock down of the clock gene Bmal1 in the SCN is associated with91
similar behavioral manifestations of depression (Landgraf et al., 2016).92
Taken together, these findings have led to the hypothesis that the disruption of the93
circadian system and specifically the internal phase coordination of circadian rhythms by94
the master circadian clock may constitute part of the etiology of mood disorders.95
Nevertheless, this hypothesis has not yet been tested in genetically and neurologically96
intact animal models—better concurring with the etiology of affective disorders in97
7. 6
humans—with targeted impairment of the master clock’s ability to maintain circadian98
internal synchronization. de la Iglesia et al. (2004) developed a genetically, neurally and99
pharmacologically intact rat model of 'forced desynchrony', in which the mere exposure100
to a symmetric 22 h light-dark (LD) cycle (LD22) leads to the uncoupling of two distinct101
populations of neuronal oscillators within the SCN: the ventrolateral SCN (vlSCN) and102
the dorsomedial SCN (dmSCN). Forced desynchronized rats present key properties of103
human forced desynchrony such as the desynchronized circadian regulation of104
temperature, sleep stages, melatonin and cortisol (Cambras et al., 2007; Lee et al., 2009;105
Schwartz et al., 2009; Wotus et al., 2013). The purpose of the present study was to take106
advantage of the forced desynchronized rat to test the prediction that a systematic107
challenge to the central circadian pacemaker’s ability to maintain the internal108
synchronization of circadian rhythms will itself cause an altered affective state. We show109
that forced desynchrony of circadian rhythms in rats induces signature symptoms that110
characterize depression in animal models including anhedonia, sexual dysfunction, and111
increased immobility in the forced swim test (FST), as well as changes in the levels and112
turnover rates of monoamines within the prefrontal cortex (PFC).113
MATERIALS AND METHODS114
Animals and housing115
Adult Wistar male rats (2-months old, Charles River, Raleigh, NC) were housed in116
individual transparent cages with ad libitum access to standard rodent diet (5001,117
LabDiet, St. Louis, MO) and water. Animals were kept in a ventilated chamber for 9-12118
weeks at constant humidity and temperature (21±1°C) under a 12:12 LD (12 h light, 12 h119
8. 7
dark) cycle (Control group, LD24) or under an 11:11 LD (11 h light, 11 h dark) cycle120
(Desynchronized group, LD22). Light intensity, as measured inside the individual cages121
at the level of the rat’s eyes, was 64 ± 11 lux during the light phase and < 1 lux dim red122
light during the dark phase. Animals were individually housed because group housing123
interferes with desynchrony and the recording of locomotor activity rhythm of each124
individual. In an effort to minimize stress during the behavioral tests, rats were briefly125
handled 2-4 times a week during the dark phase by the same experimenter that performed126
the behavioral tests. Females were not used for behavioral tests (except for testing male127
sexual behavior) because the interaction between circadian desynchrony and estrous128
cycle, which is expected to affect behavioral tests, would require more animals and a129
different experimental design. The Institutional Animal Care and Use Committee of the130
University of Washington approved all procedures.131
Forced desynchrony132
Desynchrony was assessed by continuous recording of locomotor activity rhythms133
through crossed infrared beams and analysis with the Sokolove and Bushell (1978)134
periodogram. Infrared beam breaks were recorded in 10-min bins using Clocklab135
(Actimetrics, Wilmette, IL). After three weeks under LD22, two locomotor rhythms136
emerged with different period lengths, confirmed by Sokolove-Bushel periodogram137
analysis using El Temps (A. Díez-Noguera; Universidad de Barcelona, Barcelona; Fig.138
1). One rhythm was entrained by the LD cycle and had a period of 22 h and the other was139
dissociated from the LD cycle and had a period greater than 24 h (~25 h). Rats that failed140
to desynchronize (~20%) were removed from the study. The two locomotor activity141
rhythms of desynchronized rats periodically come in phase (Aligned days) when the 22 h142
9. 8
and ~25 h locomotor activity bouts overlap with each other (Fig. 1; circled A and B), and143
come out of phase (Misaligned days), when one locomotor activity bout starts where the144
other ends (Fig. 1; circled C and D). During aligned days, both locomotor activity bouts145
(τ = 22 h and τ = ~25 h) occur during the dark phase and both rest bouts during the light146
phase.147
Behavioral tests148
For the behavioral tests, control rats were tested in either the middle of their active149
phase (Zeitgeber time (ZT) 18, where ZT12 is the time of lights off) or the middle of their150
rest phase (ZT6). Similarly, desynchronized rats were tested on aligned days in either the151
middle of their active phase (circled A in Fig. 1) or the middle of their rest phase (circled152
B in Fig. 1), or on misaligned days in either the middle of the LD22 light phase (circled C153
in Fig. 1) or the middle of the LD22 dark phase (circled D in Fig. 1). Rats were also154
weighed to the nearest ± 0.2 g (V41PWE1501T; Ohaus, Parsippany, NJ, USA) once a155
week when their cages were changed to assess differences in body weight between156
groups as body weight changes have been linked to depression (Shioiri et al., 1993).157
We had three cohorts of rats: One cohort of animals (Control: n = 22; Desynchronized:158
n = 47) was tested in the saccharin preference test (SPT) and sexual behavior test (SBT)159
allowing a recovery period of at least a week between two sequential tests. A sample of160
rats from this cohort (Control: n = 12; Desynchronized: n = 19) was tested in the open161
field test (OFT). A second cohort of animals (Control: n = 16; Desynchronized: n = 24)162
underwent a single forced-swim test (FST). Finally, a third cohort of animals (Control:163
n = 9; Aligned: n = 8) was subjected to a single OFT followed by decapitation and164
10. 9
harvesting brain tissue for chromatographic analysis. The OFT results from this cohort165
(not shown) were similar to those from cohort 1. For chromatography, we only used rats166
during aligned days because they showed the most pronounced differences in behavioral167
performance when compared to control rats. All of the behavioral tests, with the168
exception of the ‘saccharin preference test’ that was done in the rats’ home cage, were169
done in a ventilated chamber kept at constant temperature and humidity. Animals were170
transferred to the behavioral testing room in their home cages. Lighting conditions in the171
testing chamber were similar to those in the housing chamber at the time of the test:172
approximately 60 lux for animals tested during the light phase and 1lux red light for173
animals tested during the dark phase. The latter animals were transported to the174
behavioral test chamber under darkness.175
Saccharin preference test (SPT)176
To assess anhedonia, rats from both groups were offered two identical bottles: one177
filled with tap water and the other with 0.1% saccharin solution in tap water for a period178
of 8 days. Bottles were weighed daily to ± 0.1 g to track liquid consumption of the179
animals over a period of 24 h, and the position of the bottles was switched daily (at180
15:00, which corresponds to ZT 12 for controls, and falls at different time points on the181
LD cycle of the desynchronized rats) to prevent possible effects of side preference in182
drinking. We then calculated the proportion of 0.1% sucrose consumption out of total183
liquid consumption. For LD24 animals the data from all 8 days was included. During the184
8-day period, each LD22 animal went through one aligned and one misaligned day; only185
these days were included in the analysis and days in which the animals were neither186
aligned nor misaligned were not included.187
11. 10
Open field test (OFT)188
To assess anxiety, desynchronized and control rats were placed at the lower right189
corner of a black square Plexiglas arena (90 × 90 × 30 cm; width × length × height).190
Animal movements were monitored for 10 min using a surveillance video camera (WV-191
BP334, Panasonic) attached to the ceiling, 2 m above the center of the arena. We used an192
open-source automated movement-tracking code that uses MATLAB (MathWorks) as a193
platform (http://www.seas.upenn.edu/~molneuro/autotyping.html) to calculate the194
fraction of time spent in the perimeters of the arena (Patel et al., 2014), which is a195
measure of anxiety.196
Sexual behavior test (SBT)197
Sexual dysfunction is reported in 50-60% of untreated patients with depression198
(Kennedy et al., 1999; Bonierbale and Tignol, 2003), and sexual enquiry is recommended199
as an integral component of the medical evaluation (Basson and Schultz, 2007). To assess200
sexual function, males were placed in a transparent Plexiglas arena (40 × 60 × 40 cm;201
width × length × height) covered with a 5mm layer of bedding (Bed o’Cobs, The202
Andersons, Maumee, OH) and were allowed 5 minutes to habituate to the arena before a203
receptive female was introduced. The behavior of the rat pair was monitored with a204
surveillance camera (WV-BP334, Panasonic) until the first event of ejaculation occurred205
or for 30 minutes if the male did not ejaculate. During this period, we recorded the206
frequency and timing of three different male behaviors: mounting, intromission, and207
ejaculation as previously defined by Ågmo (1997). Using this information, we calculated208
12. 11
several standard parameters of sexual behavior: latency to ejaculation, latency to209
mounting, latency to intromission, as well as copulatory rate as follows,210
( )mountinglatency tonejaculatiolatency to
onsintromissi#mountings#
rateCopulatory
−
+
=211
To induce sexual receptiveness, females were anesthetized with isoflurane by212
inhalation. Their ovaries were removed following a flank incision of about 1.5 cm, and213
another 1cm incision to the underlying muscle tissue. The oviduct was ligated with214
absorbable sutures and the ovary was then removed. The same procedure was repeated on215
the opposite side to remove the second ovary. The peritoneum was sutured with216
absorbable sutures and the skin with wound clips. Females were given at least 7 days of217
recovery before being used for the behavioral tests. Sexual receptiveness was induced by218
two injections of 20 µg of estradiol suspended in flaxseed oil 48 h and 24 h prior to the219
behavioral test and an injection of 1 mg of progesterone suspended in flaxseed oil 4 h220
prior to the behavioral test.221
Forced-swim test (FST)222
Rats were individually subjected to a unique 5min FST in a separate room following223
Gonzalez and Ashton-Jones (2008). Briefly, rats provided with a flotation aid were224
placed for 5 min in a tank (38.5 cm height × 30.5 cm diameter; INSTECH, Plymouth225
Meeting, PA) with warm water (30°C). The tank was placed inside a black cylinder to226
reduce visual stimuli during the test. Rats were briefly handled throughout the two weeks227
before the test. The brief handling, warm water and flotation aid were used to reduce228
stress as a confounding factor for the evaluation of a mood-related behavioral phenotype.229
Under these conditions, Gonzalez and Aston-Jones (2008) demonstrated that a single230
13. 12
exposure to the FST can reveal a depressive behavioral phenotype in a single assay. A231
digital video camera over the tank recorded behaviors during the FST. Immediately after232
the test, each animal was removed from the water, towel-dried, and returned to its home233
cage. The water was changed and the cylinder was cleaned between rats. Two observers234
blinded to the experimental conditions scored immobility (when the animal is being235
completely motionless, with no movements of limb, tail, or head) off-line on a video236
monitor from videotaped images using Behavior Coding 2.18 (David Hurley, University237
of Washington).238
High-pressure liquid chromatography (HPLC)239
Immediately following the OFT (the only test these animals underwent), LD24 control240
rats and LD22 aligned rats were moved to an adjacent room with the same lighting241
conditions of the test, where they were decapitated with a guillotine. The brain was242
rapidly removed from the skull and placed on filter paper soaked in 0.01 M chilled243
phosphate-buffered saline. A single coronal cut was made across both hemispheres of the244
brain at Bregma + 3.7 mm and both the left and right PFC regions (Paxinos and Watson,245
1998) were dissected with fine scissors. The tissue was placed in a centrifuge tube and246
quickly frozen on dry ice. Less than two minutes passed from the time of decapitation to247
the freezing of the prelimbic and cingulate cortex tissue. The tissue was stored at -80°C248
until processing.249
Each tissue sample was sonicated in 0.5 ml of 0.1 M perchloric acid and 2 μM250
ascorbic acid. A 100 μl aliquot of the sonicated material was stored at -70ºC for protein251
determination with a Pierce BCA™ Protein Assay Kit (Thermo Scientific, Rockford, IL).252
After centrifugation of the remainder of the sonicated material at 13,000 g for 15 min, the253
14. 13
supernatant was collected and stored at -70°C for later use. After thawing, 200 μl of the254
supernatant was filtered through a 0.22 Millex GV syringe-driven filter and transferred to255
an autosampler tube prior to injection. Detection was performed with an ESA Coulochem256
II electrochemical detector (ESA, Chelmsford, MA, USA) with the conditioning cell set257
at -300 mV, electrode 1 of the analytic cell set at -90 mV, and electrode 2 of the analytic258
cell set at +350 mV, and a Phenomonex reverse phase c18 Gemini column (150 x259
4.6 mm, 3µm, 110 Å) (Phenomonex, Torrance, CA). The EZChrom Elite™260
chromatography data system (Agilent Technologies, Inc., Santa Clara, CA) was used for261
data reduction. Norepinephrine (NE), 3-methoxy-4-hydroxyphenylglycol (MHPG),262
dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), serotonin (5-HT) and 5-263
hydroxyindole acetic acid (5-HIAA) were separated and quantified. Concentrations are264
expressed as pg/µg protein.265
Statistical analysis266
The change in body mass and mean locomotor activity of rats from different groups267
were compared using repeated measures ANOVA with rat ID as a random factor nested268
within treatment group. Many behavioral tests are heavily reliant on locomotor activity.269
To account for this confounding effect, we included mean locomotor activity in the home270
cage of each rat during the day of the test as a covariate. Whenever locomotor activity did271
not have a significant effect on behavioral performance it was excluded from the model,272
and we report its effect only when found significant. Fraction of saccharin consumption273
was not normally distributed, and therefore we used the non-parametric Kolmogorov-274
Smirnov test to compare the groups, and used Bonferroni correction for multiple275
comparisons. The reported p-values are corrected values. Fraction of time spent in276
15. 14
perimeter in the OFT, immobility during the FST, copulatory rate and levels of277
monoamines in the PFC were all compared between groups and phase using two-way278
ANOVA. Because a significant number of animals did not display sexual behavior during279
the test, our data were strongly right skewed. For this reason, we used Kaplan-Meier280
survival analysis to compare differences in latency to mounting, intromission and281
ejaculation between desynchronized and control rats during the dark and light phases.282
We also calculated a depression score for rats that were tested on multiple behavioral283
tests. Because the forced swim test was done on a different cohort of rats it was not284
included. Thus, the depression score we calculated for each individual (Control: n = 20,285
Desynchronized: n = 47) was the sum of the standardized (as z-values obtained286
subtracting the mean and dividing by the standard deviation) mounting latency,287
standardized ejaculation latency, standardized copulatory rate, and standardized saccharin288
consumption. We multiplied the standardized saccharin consumption and copulatory rate289
by -1 to make sure that for all behavioral measures, a higher score is indicative of a290
higher depressive phenotype. Because the behavioral scores did not distribute normally291
we used the Kruskal Wallis non-parametric test, to compare the behavioral scores of the292
three different treatment groups. Finally, we used linear regression analysis to test the293
relationship between performance in the different behavioral tests and the amplitude of294
locomotor activity rhythm on the day of the test, calculated as the difference between295
mean locomotor activity per hour during the dark and light phases. In the case of296
saccharin preference, we tested this correlation using values from a single day. Because297
saccharin consumption was measured every 24 h over several days, LD22 rat data are298
divided into aligned and misaligned at the end of the experiment by compiling days of299
16. 15
alignment or days of misalignment for each animal. Tukey’s post hoc test was used for300
pairwise multiple comparisons when groups or the interaction were found statistically301
different. All statistical tests were done with R version 3.1.2 using ‘nlme’, ‘R.matlab’ and302
‘survival’ packages.303
RESULTS304
Body mass and locomotor activity305
Body mass of control rats (372.75 ± 15.19 g, n = 22) did not significantly differ from306
that of desynchronized rats (382.17 ± 10.36 g, n = 47; Fig. 2A; F1,67 = 3.57, p = 0.06). As307
expected, the circadian rhythm of locomotor activity significantly differed between308
groups and phase of day (Fig. 2B,C; interaction effect - F2,161 = 165.9, p < 0.001).309
Specifically, during the light phase LD24 control rats had lower locomotor activity310
(n = 22) than LD22 rats during aligned days (n = 47; Tukey’s post hoc test: p < 0.001),311
and LD22 misaligned rats had the highest locomotor activity levels (n = 47; Tukey’s post312
hoc test: p < 0.001). During the dark phase, there was no difference between locomotor313
activity levels of LD24 control rats (n = 22) and LD22 aligned rats (n = 47; Tukey’s post314
hoc test: p = 0.9), but LD22 misaligned rats had significantly lower levels of locomotor315
activity (n = 47; Tukey’s post hoc test: p < 0.001). Finally, the amplitude of locomotor316
activity was significantly different between groups (F2,46 = 130.5, p < 0.001), where317
LD24 control rats had the highest amplitude of locomotor activity, and LD22 misaligned318
rats had the lowest amplitude compared to both LD22 aligned rats and LD24 control rats.319
Circadian forced desynchrony and behavioral performance320
17. 16
Saccharin Preference Test (SPT): We did not find a difference in saccharin preference321
throughout the 8 days of the test (F1,257 = 2.38, p = 0.1). This allowed us to consolidate322
data of LD22 desynchronized rats measured on different days, as alignment and323
misalignment days did not fall on the same date for each individual. LD24 control rats324
consumed a significantly larger fraction of 0.1% saccharin (0.76 ± 0.02, n = 22) than325
LD22 rats measured during aligned days (0.65 ± 0.03, n = 47; Fig. 3A; D = 0.19, p =326
0.03), but not more than LD22 rats measured during misaligned days (0.69 ± 0.04,327
n = 47; D = 0.15, p = 0.6). Saccharin consumption did not differ between rats measured328
during aligned and misaligned days (Fig. 3A; D = 0.14, p = 0.5).329
Open Field Test (OFT): No differences were found between treatment groups in the330
fraction of time rats spent in the perimeter of the open field test arena (Fig. 3B; group:331
F2,25 = 2.52, p = 0.1; group×phase interaction: F2,25 = 0.86, p = 0.4), and there was no332
difference in preference for the perimeter between the light and the dark phases (Fig. 3B;333
phase: F1,25 = 0.12, p = 0.7). In addition, no differences were found in the distance334
travelled by the rats (group: F2,25 = 1.49, p = 0.2, group×phase interaction: F1,25 = 0.46,335
p = 0.6), but the distance travelled by all rats during the dark phase was greater than the336
distance travelled by all rats during the light phase (phase: F1,25 = 18.12, p < 0.001).337
Sexual Behavior test (SBT): Circadian forced desynchrony had an effect on sexual338
behavior. During the dark phase (Fig. 4A; χ2
= 7.83, p = 0.02) LD24 control rats (n = 11)339
had a shorter mounting latency than both aligned (n = 11; Tukey’s post hoc test: p = 0.01)340
and misaligned rats (n = 12; Tukey’s post hoc test: p = 0.003), but no such differences341
were found in the light phase (χ2
= 2.46, p = 0.3). Similarly, a difference in ejaculation342
latency was found during the dark phase (Fig. 4B; χ2
= 7.89, p = 0.02), with LD24 control343
18. 17
rats (n = 11) having shorter ejaculation latency than LD22 aligned (n = 11; Tukey’s post344
hoc test: p = 0.02) but not misaligned (n = 12; Tukey’s post hoc test: p = 0.06) rats. When345
we included mean locomotor activity as a covariate, we found that it had a significant346
effect on ejaculation latency (F1,56 = 4.20, p = 0.05), but this effect did not alter the results347
because the effect of group remained significant (F2,56 = 4.85, p = 0.01). No differences in348
ejaculation latency were found during the light phase (χ2
= 3.70, p = 0.2). Furthermore,349
no difference was found in intromission latency during the light (χ2
= 2.79, p = 0.3), or350
the dark phases (χ2
= 2.11, p = 0.3). Finally, LD22 aligned rats had lower copulatory rate351
than LD24 control rats during the dark phase (Fig. 4C; F2,62 = 4.71, p = 0.01), whereas352
misaligned rats did not differ from aligned (Tukey’s post hoc test: p = 1.0) or LD24353
control rats (Tukey’s post hoc test: p = 0.07). Mean locomotor activity had a significant354
effect on copulatory rate (F1,56 = 9.65, p = 0.003), but even when this effect was included355
in the model, we found a significant effect of group (F2,56 = 3.63, p = 0.03).356
Because females were tested at different time points during their 24 h cycle, we357
included the female Zeitgeber time as a covariate in our analysis. We found no effect of358
the female Zeitgeber time on neither ejaculation latency (F1,66 = 2.39, p = 0.1), nor359
mounting latency (F1,66 = 1.51, p = 0.2). We did find a significant effect of the female360
zeitgeber time on copulatory rate (F1,66 = 3.99, p = 0.05). However, when accounting for361
this effect, we still find a significant difference between groups (group: F2,56 = 3.49, p =362
0.04; group×phase interaction: F2,56 = 4.30, p = 0.02), with LD24 control rats showing363
higher copulatory rate during the dark phase compared to LD22 aligned rats (Tukey’s364
post hoc test: p = 0.05), but not compared to LD22 misaligned rats (Tukey’s post hoc365
test: p = 0.1). Copulatory rate of LD22 misaligned rats was also not statistically different366
19. 18
from that of LD22 aligned rats (Tukey’s post hoc test: p = 0.99). Thus, the effects were367
identical to those we report above.368
Forced Swim Test (FST): Immobility time during the FST was significantly different369
between treatment groups (Fig. 3C; F2,34 = 8.42, p = 0.001). Specifically, during the dark370
phase LD24 control rats showed significantly lower immobility time than LD22 aligned371
(Tukey’s post hoc test: p = 0.02) and misaligned rats (Tukey’s post hoc test: p = 0.02).372
During the light phase LD24 rats had significantly higher immobility time than LD22373
aligned rats (Tukey’s post hoc test: p = 0.01) and misaligned rats (Tukey’s post hoc test:374
p = 0.05).375
Depression score: We found that there is a significant difference between scores of376
different groups (Fig. 5A; χ2
(2) = 7.63, p = 0.02). Specifically, the depression score of377
control rats (-0.43 ± 0.27) was significantly lower than that of LD22 aligned (0.89 ± 0.45;378
post hoc test: p = 0.03) as well as LD22 misaligned rats (0.84 ± 0.42; post hoc test: p =379
0.03).380
Correlations between amplitude of locomotor activity and performance in the381
behavioral tests: The fraction of 0.1% saccharin consumption over a period of 24 h was382
positively correlated with the amplitude of locomotor activity during the day of the test383
(Fig. 5B; F1,69 = 10.17, p = 0.002, R2
= 0.13). Additionally, both latency to mounting384
(Fig. 5C; F1,33 = 18.29, p < 0.001, R2
= 0.36) and latency to ejaculation (Fig. 5D; F1,25 =385
6.29, p = 0.02, R2
= 0.20) in the dark phase were negatively correlated with the amplitude386
of locomotor activity during the day of the test. Finally, we found a significant positive387
20. 19
correlation between immobility in the FST and the amplitude of locomotor activity388
during the day of the test (Fig. 5E; F1,7 = 5.60, p = 0.05, R2
= 0.44).389
Circadian forced desynchrony and prefrontal cortex monoamine levels and390
turnover391
We analyzed the amount of NE, DA and 5-HT and their respective metabolites in the392
PFC at the same phases in which behavioral tests were performed in LD24 and LD22393
aligned animals (Fig. 6). Table 1 shows monoamine and metabolite levels, their394
calculated turnover and statistical results. NE (Fig. 6A; F1,28 = 4.49, p = 0.04) and DA395
levels (Fig. 6B; F1,28 = 10.21, p = 0.003) were higher in LD22 desynchronized rats396
compared to LD 24 controls (25% and 48%, respectively), and neither group showed a397
circadian oscillation in NE (Fig. 6A; F1,28 = 1.10, p = 0.3) or DA levels (Fig. 6B;398
F1,28 = 2.00, p = 0.2). 5-HT levels were higher during the dark than during the light phase399
in both groups (Fig. 6C; F1,15 = 4.60, p = 0.05), and there were no differences in 5-HT400
levels between LD24 controls and LD22 desynchronized rats (Fig. 6C; F1,15 = 0.24,401
p = 0.6). Finally, there were no differences in turnover rates of NE (Fig. 6A’; F1,28 = 0.89,402
p = 0.4) or DA (Fig. 6B’; F1,28 = 0.0006, p = 0.98) between groups, but turnover rate of 5-403
HT was significantly higher in LD22 desynchronized rats than in LD 24 controls (Fig.404
6C’; F1,15 = 12.80, p = 0.003).405
DISCUSSION406
The link between affective disorders and disruptions of the circadian system has been407
recognized for decades (Wulff et al., 2010; McClung, 2013). Nevertheless, it is unclear408
whether circadian disorders may represent a comorbidity of mental disease or,409
21. 20
alternatively, challenges to circadian organization may be among the prevalent factors410
that increase susceptibility to mood disorders. Our results show that long-term411
desynchronization of the circadian system in a neurally, genetically and412
pharmacologically intact mammal leads to depressive-like behavior, as indicated by413
increased immobility in the FST and decreased hedonic behavior in the SPT and SBT414
(Figs. 3 and 4). Moreover, we found that the severity of the circadian desynchronization,415
as estimated by the amplitude of locomotor activity, was correlated with performance in416
the behavioral tests (Fig. 5). Specifically, the lower the amplitude of locomotor activity417
rhythm, the stronger the depressive phenotype was on the behavioral tests. These results418
suggest that internal circadian desynchronization contributes to the pathogenesis of419
affective disorders.420
The manipulation of the LD cycle in the forced desynchronized rat leads to the421
desynchronization of circadian rhythms, including the rhythms of core body temperature,422
release of corticosterone and melatonin, and REM sleep (de la Iglesia et al., 2004;423
Cambras et al., 2007; Lee et al., 2009; Schwartz et al., 2009; Wotus et al., 2013). The424
internal misalignment of these circadian rhythms is characteristic of depression in425
humans as well (Souêtre et al., 1989; Riemann et al., 2001; Lewy, 2009). This426
misalignment is in agreement with the pattern of clock gene expression in extra-SCN427
neural circadian oscillators in in post mortem tissue from human patients with depression428
and in animal models of depression (Li et al., 2013; Landgraf et al., 2015; Logan et al.,429
2015; Savalli et al., 2015). However, depression in the animal models used in these430
studies was induced by aversive methods, and the causality between desynchronization431
and the manifestation of depression could not be established. In the forced432
22. 21
desynchronized rat, the simple manipulation of the LD cycle causes the predictable and433
stable desynchronization of neuronal oscillators within the SCN in the absence of any434
invasive intervention, and this in turn leads to a depressive behavioral phenotype.435
For the present study we used a single-exposure FST that has been previously436
validated in an animal model of light deprivation-induced depression. Rats kept for 6437
weeks in constant darkness showed increased immobility in the first 5 min of exposure to438
the test compared to controls, and this behavioral deficit was reversed by desipramine439
(Gonzalez and Aston-Jones, 2008). Similarly, desynchronized rats during the dark phase440
in our study showed a spontaneous despair behavior revealed by the increased immobility441
during the 5-min FST, an expected outcome if circadian desynchrony predisposes442
individuals to a behavioral depressive phenotype. Importantly, the FST depressive443
phenotype observed in aligned LD22 animals during the dark phase, as compared to444
LD24 animals during the same phase, cannot be attributed to reduced locomotor activity445
because the activity in the home cage for these groups did not differ (Fig. 2B,C). We446
argue that this reduced response—increased immobility—to helplessness during a time of447
the day at which the animals are expected to be more active represents a phenotypic448
manifestation of depression.449
Control rats kept under LD24, showed a daily (likely circadian) modulation of450
performance in the FST, manifested as less immobility during the dark phase. This451
modulation has been largely ignored in studies using behavioral markers of depression,452
which typically perform the tests during the rest (light) phase of the animals. Indeed,453
Tataroğlu et al. (2004) have shown that bilateral lesion of the SCN in rats leads to454
reduced behavioral despair, and suggested that the lesion had an ameliorative effect on455
23. 22
the strategy to respond to the stress of the FST. Because the animals in this study were456
tested during the light phase, an alternative explanation that we favor for this result is that457
the lesions do not reduce despair but instead eliminate the circadian modulation of the458
FST performance.459
In contrast to LD24 rats, LD22 animals did not show similar circadian modulation of460
FST performance, as evident from the lack of differences in levels of immobility between461
the dark and light phases. Specifically, rats tested during both aligned and misaligned462
days showed higher immobility during the dark phase and lower immobility during the463
light phase compared to controls. Because mood was shown to oscillate daily in humans464
from different cultures around the globe (Golder and Macy, 2011) and a blunted rhythm465
of mood during the 24 h day was found in depressive patients (Boivin, 2000; Wirz-466
Justice, 2006), we suggest that LD22 aligned rats show attenuated rhythm of immobility467
in the FST, characterized by respectively lower and higher immobility during the light468
and dark phases than controls, and no difference in levels of immobility between the two469
phases.470
Strikingly, we found that LD22 desynchronized rats displayed lower levels of471
immobility during the light phase than LD24 control rats (Fig. 3C). We do not think that472
this is an indication of manic behavior for two reasons. First, LD22 rats consistently473
display anhedonia in the other behavioral tests. Second, because misaligned animals are474
tested in conflicting phases—subjective day during the dark phase or subjective night475
during the light phase—their immobility in the FST could reflect the opposing effects of476
light and the circadian system on escape performance rather than the depressive state of477
the animal. In line with this interpretation, immobility in LD22 misaligned animals478
24. 23
during the light phase was higher than immobility in LD24 animals during the dark phase479
but lower than immobility in in LD24 animals during the light phase.480
Our study is the first to show that internal circadian desynchronization leads to sexual481
dysfunction. We found that circadian desynchronization affects both sexual motivation,482
estimated by the latency to mount (Pfaus et al., 1990), and sexual function, estimated by483
the latency to and occurrence of ejaculation (Ågmo, 1997). Moreover, the severity of the484
circadian disturbance, i.e. the reduction in amplitude of locomotor activity during the day485
of the test, was correlated with both measures (Fig. 5C,D), suggesting that the486
impairment of sexual behavior is linked to the extent of circadian disruption. The487
prevalence of sexual dysfunction in depressive patients not treated with antidepressants is488
estimated to be 50-60%, but this value is likely to be an underestimation because sexual489
enquiry is often not included in the medical assessment (Kennedy et al., 1999; Bonierbale490
and Tignol, 2003; Basson and Schultz, 2007). Our results suggest that circadian491
desynchronization may be a contributing factor to both negative affect and sexual492
dysfunction. Of note, sexual dysfunction has not been previously tested in animal models493
of depression, our results show that it represents a sensitive behavioral outcome to assess494
the depressive phenotype of an animal, and one that is more meaningful than other495
metrics both for its ecophysiological and translational value. Specifically, the FST has496
been criticized as a behavioral assessment for depression because (1) it is heavily497
dependent on locomotor activity performance (Slattery and Cryan, 2012) and (2) as a498
method to induce depression, it represents an unusually stressful situation that bares little499
relationship to depression triggers in humans (Holmes, 2003). In the present study, we500
did not use the FST to induce depression but rather as a probe test to assess a preexisting501
25. 24
depressive state. Nevertheless, it still can be argued that a test in which an individual has502
to swim after being placed in an unescapable pool of water is unlike most stressful503
situations that lead to a manifestation of depression in humans. In this context, we504
propose that assessing sexual performance may represent a more sensitive and505
meaningful testing approach in animal models of depression.506
The depressive-like phenotype we found was independent of levels of anxiety, as507
evident from the lack of differences between desynchronized and control preference for508
the perimeter of the arena in the OFT (Fig. 3B). Although there is a strong comorbidity509
between anxiety disorders and depression, they are still considered independent disorders.510
This suggests that the desynchronized rat may serve as a model for exploring depressive511
behavior independently of anxiety, although further tests of anxiety level are required to512
confirm this finding.513
The correlation between abnormal light exposure, disrupted rhythms and mood514
disorders suggests that photic stimulation of the circadian system through the RHT is515
critical for the maintenance of mood. This mechanism could explain seasonal affective516
disorder (SAD), a form of depression linked to the short winter photoperiod that is517
responsive to bright light therapy (Golden et al., 2005). The circadian basis of light518
therapy-induced improvement in mood in SAD patients is supported by the fact that light519
appears to be more beneficial when applied at a specific circadian phase: for some520
patients morning treatment is efficacious and for others it is evening treatment (Terman521
and Terman, 2005). Furthermore, light therapy is beneficial not only in SAD but in522
virtually all forms of depression, suggesting that circadian pathologies could contribute to523
the etiology of depression in general (Pail et al., 2011). Remarkably, nocturnal rats524
26. 25
present an anatomical and behavioral depressive phenotype after weeks of exposure to525
total darkness. The absence of a light alone reduces the amplitude of the SCN-dependent526
sleep-wake circadian rhythm and generates neural damage in monoamine brain systems527
that are SCN-transynaptic targets, in association with a depressive behavioral phenotype528
(Gonzalez and Aston-Jones, 2006, 2008). Interestingly, Dulcis et al. (2013) showed that529
the type of neurotransmitter released from nuclei receiving direct SCN input is dependent530
on photoperiod length, and that the alteration in neurotransmitter release is associated531
with a depressive-like behavior in mice. Forced desynchronized rats, however, are not532
exposed to different amounts of light, as the LD22 schedule results in the same533
proportion of light and dark as an LD24 schedule. Thus, the depressive behavioral534
phenotype of forced desynchronized rats likely emerges from the stable desynchrony535
between the vl- and dmSCN. This appears to differ from the depressive phenotype that536
emerges in mice exposed to aberrant LD cycles of 7 h (3.5L:3.5D), in which the SCN537
appears not to be affected by the schedule and the phenotype has been proposed to be538
mediated by retinal innervation of limbic regions (LeGates et al., 2012; LeGates et al.,539
2014).540
The monoamine hypothesis of depression postulates that the pathogenesis of541
depression involves impairment of 5-HT, NE and DA systems in the central nervous542
system (Delgado, 2000; Hirschfeld, 2000), and there is evidence that these abnormalities543
are localized to the PFC and the limbic system (Drevets, 1999). Our desynchrony544
protocol leads to an imbalance in the regulation of monoamine levels and their release in545
the PFC, which is manifested as higher levels of NE and DA and higher 5-HT turnover546
rate (Fig. 6). The role of DA in depression is still not well understood, and is challenged547
27. 26
by the paradoxical antidepressant effect of both agonists and antagonists of DA (Dailly et548
al., 2004). In stressed rats, there is increased release of NE and DA in the PFC, and549
chronic mild stress causes an increase in the magnitude of this response (Finlay et al.,550
1995; Di Chiara et al., 1999). It is therefore possible that the higher levels of NE and DA551
we found in the PFC of LD22 desynchronized rats result from stress sensitization of these552
individuals, as brain tissue of these rats was collected immediately following an OFT. In553
contrast, we found unchanged 5-HT levels combined with higher 5-HT turnover rate in554
LD22 desynchronized rats, suggesting that these animals have a lower availability of 5-555
HT, which could account for their depressive like behavior. This is also consistent with556
decreased availability of 5-HT in patients with SAD, and the 24% decrease in557
serotonergic axons found in the PFC of suicide victims (Austin et al., 2002; Pail et al.,558
2011). Our findings provide a direct link between stable circadian desynchronization and559
an imbalance in monoamine regulation in the PFC. Obviously, the PFC is not the only560
brain region involved in the modulation of motivational states, and changes in other561
transmitter systems, as well as in intracellular regulatory pathways within or outside the562
PFC, could underlie the expression of depressive behaviors in the forced desynchronized563
rat.564
In summary, our study shows that the stable forced desynchrony of circadian rhythms565
leads to the manifestation of a depressive-like phenotype. This phenotype likely emerges566
from the desynchronization of neuronal oscillators within the SCN, and is associated with567
the severity of circadian desynchronization and an imbalance in monoamine metabolism568
within the PFC. Taken together, our findings provide direct support for a circadian569
desynchronization etiology of depressive behavior. Hence, the forced desynchronized rat570
28. 27
could represent a unique neurologically, pharmachologically and genetically intact571
animal model to study the neural basis of affective disorders and its relationship with572
internal circadian misalignment.573
ACKNOWLEDGEMENTS574
We wish to thank Camilo Silva, Jennifer Gile and Emma Morris for their help with575
surgeries and animal care. This research was supported by NIH R01MH075016 and NSF576
IOS0909716 to HOD, the Young Investigator Award from the Mental Health Research577
Association (National Alliance for Research on Schizophrenia and Depression) to578
MCCG, and the Geriatric Research, Education and Clinical Center and the Research and579
Development Service, VA Puget Sound Health Care System (CS and CWW). MBH was580
supported by the Israeli Committee for Higher Education Postdoctoral Fellowship for581
Women and by the Washington Research Foundation Funds for Innovation in582
Neuroengineering.583
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FIGURE LEGENDS756
Figure 1. Forced desynchrony of locomotor activity. (Top) Double-plotted actograms for757
locomotor activity of representative rats maintained on symmetric 24h (left; 12h light,758
12h dark) or 22h (right; 11h light, 11h dark) LD cycles. The grey shading indicates759
lights off. The black line on the LD22 plot indicates the onset of locomotor activity for760
the LD-dissociated bout of activity. Letters represent phases at which LD22761
desynchronized animals were tested. During aligned days animals were tested in the762
dark phase (A) or the light phase (B). During misaligned days animals were also tested763
in the light phase (C) or the dark phase (D). Although all four phases are shown764
schematically in the same animal, each animal was tested only once at one of these765
four phases. (Bottom) Periodograms of the time series represented in each actogram.766
Analysis revealed a single statistically significant peak for the LD24 animal and two767
statistically significant peaks for the LD22 animal. The numbers above the peaks768
indicate the period of the significant peaks in hours.769
Figure 2. The effect of forced desynchrony on body mass and locomotor activity. (A)770
Change in mean ± SE body mass in LD24 controls (n = 22) and LD22 desynchronized771
(n = 47) animals throughout the experiment. (B) Mean ± SE locomotor activity [% of772
total activity during each 24h or 22h cycle] of LD24 control rats and LD22773
desynchronized rats. Shaded grey area denotes the time when lights were turned off.774
Note that data for desynchronized animals corresponds to 11 h of day and 11 h of775
darkness. (C) Mean ± SE locomotor activity per hour (total IR beam interruptions)776
averaged over either the light or dark phase for control rats (black bars), aligned rats777
34. 33
(grey bars) and misaligned rats (white bars). Different letters denote statistically778
significant differences between groups.779
Figure 3. Effect of circadian forced desynchrony on behavioral manifestations of780
depression. (A) Fraction of 0.1% saccharin consumption averaged over eight days for781
LD24 control, LD22 aligned, and LD22 misaligned rats. Boxes outline the lower782
(25%) and upper (75%) quartiles and the line denotes the median. Error bars represent783
the 95% confidence intervals. Different letters denote statistically significant784
differences (p < 0.05). (B) Fraction of time spent in the perimeters of the open field785
test arena, during the light and dark phases in the same groups. (C) Immobility time786
during the 5 minutes forced swim test (FST) in the light and dark phases. Different787
letters denote statistically significant differences (p < 0.05). All values represent mean788
± SE.789
Figure 4. Effect of circadian forced desynchrony on sexual behavior of male rats during790
the dark phase. (A) Percentage of LD24 control and LD22 desynchronized male rats791
that did not mount and (B) that did not ejaculate as a function of the time lapsed from792
the introduction of a receptive female. *p < 0.05, **p < 0.01. Log-rank values were793
calculated based on a Kaplan-Meier survival analysis. (C) Mean ± SE copulatory rate794
in LD24 control and LD22 desynchronized males. Copulatory rate was calculated as795
the sum of mounting and intromission events divided by the time lapsed from the first796
mounting event until the first ejaculation event. Different letters denote statistical797
differences between groups (p < 0.05).798
35. 34
Figure 5. The effect of circadian forced desynchrony on behavioral performance. (A) A799
depression score (higher values indicate a depressive phenotype) for LD24 control,800
LD22 aligned, and LD22 misaligned rats. Boxes outline the lower (25%) and upper801
(75%) quartiles and the line denotes the median. Error bars represent the 95%802
confidence intervals and the black circles are outliers. Different letters denote803
statistically significant differences (p < 0.05). The amplitude of circadian locomotor804
activity predicts the severity of behavioral manifestations of depression. (B) Fraction805
of 0.1% saccharin consumption over a 24 h period in LD24 control and LD22806
desynchronized animals as a function of the amplitude of locomotor activity on the807
day of the test. Latency to mounting (C) and ejaculation (D) during the dark phase as a808
function of the amplitude of locomotor activity on the day of the test. (E) Immobility809
in the forced swim test during the dark phase as a function of the amplitude of810
locomotor activity on the day of the test, calculated as the difference between the811
mean activity per hour during the dark phase and the mean activity per hour during the812
light phase.813
Figure 6. Forced desynchronized rats show a significant increase of levels of814
norepinephrine (NE) and dopamine (DA), and of serotonin (5-HT) turnover in the815
prefrontal cortex. (A-C) Mean ± SE levels of each monoamine. (A’-C’) Mean ± SE816
turnover for each monoamine. *p < 0.05, **p < 0.01 (group effect); #p = 0.05 (phase817
effect). See Table 1 for statistics.818
819
36. 35
Table 1. Monoamine levels in PFC of LD24 controls and LD22 aligned820
desynchronized rats. Levels (pg/µg of protein) of monoamines and their metabolites are821
shown as mean ± SE. Turnover is shown as the ratio of each metabolite over each822
respective monoamine. The main effects of the Two-way ANOVA are shown.823
LD24 LD22 Two-way ANOVA
Light Dark Light Dark Group Phase Interaction
Monoamine (pg/μg
protein)
NE
4.33 ±
0.67
5.78 ±
0.70
6.48 ±
0.53
6.39 ±
0.66
F1,28 = 4.49;
p = 0.04
F1,28 = 1.10;
p = 0.3
F1,28 = 1.39;
p = 0.2
DA
1.08 ±
0.16
1.48 ±
0.13
1.82 ±
0.21
1.96 ±
0.21
F1,28 = 10.21;
p = 0.003
F1,28 = 2.00;
p = 0.2
F1,28 = 0.44;
p = 0.5
5-HT
12.00
± 1.79
17.00
± 1.13
14.86
± 0.96
15.40
± 1.13
F1,15 = 0.24;
p = 0.6
F1,15 = 4.60;
p = 0.05
F1,15 = 2.97;
p=0.1
Metabolite (pg/μg
protein)
MHPG
0.39 ±
0.06
0.37 ±
0.03
0.48 ±
0.03
0.41 ±
0.04
F1,28 = 3.03;
p = 0.09
F1,28 = 1.17;
p = 0.3
F1,28 = 0.38;
p=0.5
DOPAC
1.12 ±
0.23
0.89 ±
0.14
1.41 ±
0.14
1.19 ±
0.30
F1,27 = 2.34;
p = 0.1
F1,27 = 1.43;
p = 0.2
F1,27 = 0.01;
p=0.9
HIAA
5.25 ±
0.15
5.82 ±
0.50
10.26
± 1.24
8.22 ±
0.49
F1,15 = 10.52;
p = 0.006
F1,15 = 0.41;
p = 0.5
F1,15 = 1.30;
p = 0.3
Turnover
NE
(MHPG/NE)
0.09 ±
0.01
0.07
0.01
0.08 ±
0.01
0.07 ±
0.01
F1,28 = 0.89;
p = 0.4
F1,28 = 3.36;
p = 0.08
F1,28 = 1.53;
p = 0.2
DA
(DOPAC/DA)
1.06 ±
0.20
0.58 ±
0.09
0.86 ±
0.13
0.79 ±
0.18
F1,28 = 0.0006;
p = 0.98
F1,28 = 3.12;
p = 0.09
F1,28 = 0.20;
p = 0.2
5HT
(HIAA/5HT)
0.44 ±
0.01
0.35 ±
0.04
0.69 ±
0.06
0.54 ±
0.04
F1,15 = 12.80;
p = 0.003
F1,15 = 3.96;
p = 0.07
F1,15 = 0.22;
p = 0.6
824