This document outlines a study examining the effect of curcumin derivatives on AMPA receptor kinetics. The objectives are to identify potent and selective curcumin derivatives that inhibit AMPA receptors and characterize their effects. HEK293 cells will be transfected with GluA2 receptors to study the effects of curcumin derivative A on whole cell current, desensitization, and deactivation. Preliminary results show that derivative A decreases peak current, increases desensitization rate and extent, and increases deactivation rate, indicating it inhibits AMPA receptors. The study aims to better understand how curcumin derivatives modify AMPA receptor properties to develop new natural drugs without side effects.
The document discusses the glycine receptor, a ligand-gated chloride channel protein that is the major inhibitory neurotransmitter in the adult central nervous system. It exists as a pentameric protein composed of alpha and beta subunits that surround a central pore. Glycine binding activates the receptor, allowing chloride ion influx that hyperpolarizes the neuron. Disorders involving glycine receptor mutations can cause startle disease or non-ketotic hyperglycinemia. The receptor has many ligands but is antagonized primarily by strychnine.
Glutamatergic neurotransmission involves glutamate, the major excitatory neurotransmitter in the brain. There are two pathways for glutamate synthesis from precursors and multiple receptor types including NMDA, AMPA, KA, and metabotropic receptors. The different receptor subunits provide diversity in function. Glutamate signaling is involved in many brain pathways and clinical implications include roles in schizophrenia, Parkinson's disease, and drug mechanisms of action.
General principles of signal transduction
G Protein-coupled Receptors (GPCRs): Structure and Mechanism.
GPCRs that Regulate Adenylyl Cyclase.
GPCRs that Activate Phospholipase C.
GPCRs that Regulate Ion Channels.
GPCRs that Regulate Gene Transcription.
(1) The document discusses the history of the discovery of neurotransmitters and the role of Ramón y Cajal and Otto Loewi in determining neurons communicate via chemical messengers rather than electrical signals.
(2) It provides definitions of neurotransmitter and criteria that must be met for a substance to be classified as a neurotransmitter.
(3) Glutamate is described as the major excitatory neurotransmitter in the brain, present at high concentrations in presynaptic terminals and involved in many key pathways.
CPP-115 is a potent mechanism-based inactivator of the enzyme γ-aminobutyric acid aminotransferase (GABA-AT) that was developed to treat seizures and other neurological disorders. This study investigated CPP-115's inactivation mechanism of GABA-AT. It was found that CPP-115 undergoes enzyme-catalyzed hydrolysis of its difluoromethylene group, releasing two fluoride ions. This causes a conformational change in the enzyme, forming a tightly bound complex and inactivating the enzyme. Unexpectedly, CPP-115 does not follow the anticipated Michael addition pathway but instead utilizes a novel catalytic mechanism. This new mechanism of inactivation provides insights for
G-protein coupled receptors (GPCRs) are the largest family of cell surface receptors. Upon ligand binding, GPCRs activate intracellular G proteins that propagate signals via second messenger molecules. There are three major families of G proteins - Gs stimulates adenylate cyclase and increases cAMP, Gi inhibits adenylate cyclase and decreases cAMP, and Gq activates phospholipase C and increases intracellular calcium. Receptor tyrosine kinases (RTKs) activate intracellular signaling pathways through autophosphorylation upon ligand binding, which recruits signaling proteins containing SH2 domains. Major RTK pathways include Ras-MAPK, which regulates cell growth and proliferation.
The document discusses the glycine receptor, a ligand-gated chloride channel protein that is the major inhibitory neurotransmitter in the adult central nervous system. It exists as a pentameric protein composed of alpha and beta subunits that surround a central pore. Glycine binding activates the receptor, allowing chloride ion influx that hyperpolarizes the neuron. Disorders involving glycine receptor mutations can cause startle disease or non-ketotic hyperglycinemia. The receptor has many ligands but is antagonized primarily by strychnine.
Glutamatergic neurotransmission involves glutamate, the major excitatory neurotransmitter in the brain. There are two pathways for glutamate synthesis from precursors and multiple receptor types including NMDA, AMPA, KA, and metabotropic receptors. The different receptor subunits provide diversity in function. Glutamate signaling is involved in many brain pathways and clinical implications include roles in schizophrenia, Parkinson's disease, and drug mechanisms of action.
General principles of signal transduction
G Protein-coupled Receptors (GPCRs): Structure and Mechanism.
GPCRs that Regulate Adenylyl Cyclase.
GPCRs that Activate Phospholipase C.
GPCRs that Regulate Ion Channels.
GPCRs that Regulate Gene Transcription.
(1) The document discusses the history of the discovery of neurotransmitters and the role of Ramón y Cajal and Otto Loewi in determining neurons communicate via chemical messengers rather than electrical signals.
(2) It provides definitions of neurotransmitter and criteria that must be met for a substance to be classified as a neurotransmitter.
(3) Glutamate is described as the major excitatory neurotransmitter in the brain, present at high concentrations in presynaptic terminals and involved in many key pathways.
CPP-115 is a potent mechanism-based inactivator of the enzyme γ-aminobutyric acid aminotransferase (GABA-AT) that was developed to treat seizures and other neurological disorders. This study investigated CPP-115's inactivation mechanism of GABA-AT. It was found that CPP-115 undergoes enzyme-catalyzed hydrolysis of its difluoromethylene group, releasing two fluoride ions. This causes a conformational change in the enzyme, forming a tightly bound complex and inactivating the enzyme. Unexpectedly, CPP-115 does not follow the anticipated Michael addition pathway but instead utilizes a novel catalytic mechanism. This new mechanism of inactivation provides insights for
G-protein coupled receptors (GPCRs) are the largest family of cell surface receptors. Upon ligand binding, GPCRs activate intracellular G proteins that propagate signals via second messenger molecules. There are three major families of G proteins - Gs stimulates adenylate cyclase and increases cAMP, Gi inhibits adenylate cyclase and decreases cAMP, and Gq activates phospholipase C and increases intracellular calcium. Receptor tyrosine kinases (RTKs) activate intracellular signaling pathways through autophosphorylation upon ligand binding, which recruits signaling proteins containing SH2 domains. Major RTK pathways include Ras-MAPK, which regulates cell growth and proliferation.
Glutamate and glycine are important neurotransmitters in the central nervous system. Glutamate is the primary excitatory neurotransmitter and is synthesized from glucose or glutamine. It acts through ionotropic and metabotropic glutamate receptors. The three main ionotropic receptor subtypes are NMDA, AMPA, and kainate receptors. NMDA receptors require both glutamate and glycine to open the ion channel. Metabotropic glutamate receptors are G protein-coupled and modulate intracellular signaling pathways. Glycine is also a neurotransmitter and is required as a co-agonist for NMDA receptor activation, playing a role in excitatory neurotransmission.
This document summarizes several anticonvulsant drugs, including their mechanisms of action and metabolic pathways. It discusses how primidone is metabolized to phenobarbital, how carbamazepine and oxcarbazepine are metabolized by cytochrome P450 enzymes, and how gabapentin and pregabalin are not metabolized. It also provides information on the metabolic pathways and side effects of other anticonvulsants such as lamotrigine, topiramate, zonisamide, levetiracetam, tiagabine, ethosuximide, and vigabatrin. The document concludes by mentioning the uses and metabolic pathways of clonazepam,
This document discusses glutamate receptors, including their history, types, roles, and drugs that act on them. It notes that glutamate is the major excitatory neurotransmitter in the central nervous system. There are two main types of glutamate receptors: ionotropic receptors which are ligand-gated ion channels including NMDA, AMPA, and kainate receptors, and metabotropic G protein-coupled receptors divided into groups 1, 2, and 3. The roles of glutamate receptors include synaptic plasticity, learning and memory, and excitotoxicity. Many drugs have been developed that act as agonists or antagonists at glutamate receptors and are being investigated for conditions like Alzheimer's, Parkinson
This document summarizes 5 major categories of transducer mechanisms:
1) G-protein coupled receptors which activate downstream effectors like adenylyl cyclase or phospholipase C.
2) Ion channel receptors which directly open or close ion channels.
3) Transmembrane enzyme-linked receptors which activate intracellular protein kinases.
4) Transmembrane JAK-STAT binding receptors which activate the JAK/STAT signaling pathway.
5) Receptors regulating gene expression which bind intracellularly to directly regulate gene transcription.
- GABA is the major inhibitory neurotransmitter in the mammalian brain. It acts through GABAA, GABAB, and GABAC receptors.
- GABAA receptors are ligand-gated chloride channels modulated by drugs like benzodiazepines, barbiturates, and general anesthetics. GABAB receptors are G-protein coupled receptors that inhibit neurotransmitter release and hyperpolarize neurons.
- Drugs that enhance GABAergic transmission through GABAA receptors like benzodiazepines are used as sedatives, anxiolytics, and anticonvulsants. The GABAB agonist baclofen is used as a muscle relaxant for spastic
1. G-proteins bind GTP and control intracellular signaling pathways. They exist in two states - active when bound to GTP and inactive when bound to GDP.
2. G-proteins are tightly regulated by accessory proteins that modulate their cycling between GTP-bound and GDP-bound states.
3. The heterotrimeric G-proteins transmit signals from cell surface receptors to enzymes and channels. They are stimulated by receptors, act on effectors, and are regulated by nucleotide exchange and hydrolysis.
This document summarizes information about the neurotransmitters glutamate and GABA. It discusses their roles, receptors, and involvement in various neurological conditions and disorders. Glutamate is the primary excitatory neurotransmitter in the brain, while GABA is the primary inhibitory neurotransmitter. The document outlines their synthesis, metabolism, receptor types, and relationships. It also explores how imbalances in glutamate and GABA are implicated in conditions like anxiety, depression, addiction, and neurodegenerative diseases.
G Protein Coupled Receptors (GPCRs) and CancerDayananda Salam
GPCRs play an important role in many types of cancer through their activation by various ligands. Some key points:
- GPCRs make up over 1% of the human genome and are the target of over 50% of drugs. They signal through G proteins and regulate many important cellular functions.
- In cancer, GPCRs activated by ligands like LPA, SDF1, chemokines promote processes like growth, migration, angiogenesis and metastasis. Examples include LPA1 in breast cancer and CXCR4 in ovarian cancer.
- GPCRs can also interact with other pathways like EGFR to enhance signaling. Orphan GPCRs without known ligands may play emerging roles in cancers like GPR49
Phenytoin is metabolized into reactive intermediates like arene oxide and catechol that can bind to proteins and trigger immune responses. These metabolites are deactivated by enzymes like epoxide hydrolase and glutathione transferase. Phenytoin strongly induces several CYP450 isozymes and drug metabolizing enzymes. Carbamazepine is metabolized by CYP3A4 into an epoxide metabolite suspected of causing idiosyncratic reactions. Gabapentin and pregabalin modulate calcium influx and stimulate GABA biosynthesis without being metabolized. Felbamate undergoes hydroxylation and hydrolysis to form a toxic metabolite, 2-phenylpropanal. Future anticonvulsants
GABA is the primary inhibitory neurotransmitter in the central nervous system. It acts on three main receptor types: GABAA, GABAB, and GABAC. GABAA is a ligand-gated ion channel whose activation allows chloride ion influx. GABAB is a G-protein coupled receptor whose activation opens potassium channels and closes calcium channels. Anti-epileptic drugs can act on these GABA receptors and neurotransmitter pathways. Newer anti-epileptics discussed include lamotrigine, gabapentin, topiramate, levetiracetam, zonisamide, tiagabine, and vigabatrin. Their mechanisms of action involve effects on sodium channels,
Dr. Pavani discusses G protein-coupled receptors (GPCRs), which are integral membrane proteins that sense molecules outside the cell and activate internal signal transduction pathways and cellular responses. There are over 800 GPCRs in humans that detect a wide range of ligands and are involved in many physiological processes. GPCRs work by coupling to G proteins, which activate various intracellular effectors like adenylyl cyclase, phospholipase C, and ion channels. Dysregulation of GPCR signaling can lead to many human diseases. Martin Rodbell and Alfred Gilman were awarded the 1994 Nobel Prize in Physiology or Medicine for their discoveries related to G proteins and GPCR signal transduction.
Assignment on Secondary messengers and intracellular signalingDeepak Kumar
Assignment on Secondary messengers: cyclic AMP, cyclic GMP, calcium ion, inositol 1,4,5- trisphosphate, (IP3), NO, and diacylglycerol. Detailed study of following intracellular signaling pathways: cyclic AMP signaling pathway, mitogen-activated protein kinase (MAPK) signaling, Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling pathway.
The document discusses neurotransmission and non-adrenergic, non-cholinergic neurotransmission. It describes how neurotransmitters are synthesized, packaged into vesicles, released into the synaptic cleft upon neuronal stimulation, and bind to receptors on the postsynaptic neuron. It notes that some neurons release multiple neurotransmitters, including glutamate, ATP, nitric oxide, and peptides. The presentation focuses on glutamate as a major excitatory neurotransmitter in the central nervous system that acts through ionotropic AMPA, kainate, and NMDA receptors and metabotropic receptors. Disorders associated with glutamate dysregulation and its role in memory and learning are also mentioned.
The document summarizes G protein-coupled receptor (GPCR) signaling. It discusses how different GPCRs are coupled to different G proteins that activate various downstream effectors like adenylyl cyclase, phospholipase C or ion channels. It provides examples of specific GPCRs like beta-adrenergic receptors and muscarinic acetylcholine receptor and their signaling mechanisms. It also describes the role of second messengers like cAMP, IP3 and DAG in GPCR signaling.
This document discusses a study examining the interaction between dopamine and histamine in the basal ganglia. It provides background information on synaptic transmission, the histaminergic system, dopamine signaling in the striatum, and Parkinson's disease. The study aims to investigate how activation of dopamine D1 receptors regulates phosphorylation of glutamate AMPA receptors and DARPP-32 in striatal slices, and how histamine H3 receptor activation may oppose these effects of dopamine signaling. The results suggest dopamine increases phosphorylation of AMPA receptors and DARPP-32, while histamine H3 receptor activation reduces dopamine-induced phosphorylation, indicating opposing regulation of cAMP/PKA signaling by dopamine and histamine in striatal neurons.
G protein-coupled receptors (GPCRs) are seven transmembrane receptors that activate intracellular signaling pathways through heterotrimeric G proteins. When an agonist binds to a GPCR, it causes a conformational change activating the G protein's α subunit. The activated α subunit then interacts with downstream effectors like adenylyl cyclase or phospholipase C. This leads to the production of second messengers like cAMP or IP3. Mutations in GPCRs and G proteins can cause disease by disrupting signaling, with loss or gain of function mutations associated with various endocrine disorders and cancers.
This document compares two methods for sequencing proteins: Edman degradation and mass spectrometry. Edman degradation involves tagging and removing amino acids from the N-terminal end one by one to determine the sequence. Mass spectrometry involves digesting proteins into peptides, using mass to charge ratios to separate peptides, and fragmenting peptides to measure fragment ion masses and deduce sequences. Mass spectrometry is more sensitive, does not require purified samples, and can handle modified proteins better than Edman degradation.
Alcohol related changes in the regulation of NMDA receptor functions-József NagyNiyamat Chimthanawala
A concise review about all the events supposedly playing a
regulatory role in the up-regulation of NMDAR functions in
consequence of chronic ethanol exposure.
The effect of subunit expression, molecular mechanisms, protein kinase/phosphatases activity, post-synaptic density, allosteric modulators activity is observed.
G Protein–Coupled Receptors (GPCRs) are integral membrane proteins that are activated by extracellular signaling molecules and activate intracellular secondary messenger pathways. They have seven transmembrane domains and activate heterotrimeric G proteins upon ligand binding. The G protein then activates downstream effector enzymes like adenylyl cyclase, which generates secondary messengers like cAMP. These messengers go on to activate pathways that ultimately alter cell function. The beta-adrenergic receptor pathway is a key example, with epinephrine binding and cAMP production leading to protein kinase A activation. PKA then phosphorylates target proteins to produce effects like increased heart rate. GPCRs are major drug targets, and their deregulation
Neurohumoral transmission in CNS ,special emphasis on importance of various neurotransmitters like with GABA, Glutamate, Glycine, serotonin and dopamine
The document discusses second messenger systems. It describes how second messengers relay signals from cell surface receptors to target molecules inside the cell. Some key points discussed include:
- Earl Sutherland discovered cyclic AMP (cAMP) as the second messenger for epinephrine and won the Nobel Prize for this work.
- Common second messenger systems include those using cAMP, cGMP, phosphatidylinositol, and tyrosine kinases as secondary messengers.
- G proteins act as transducers between receptors and effectors and are important drug targets.
- cAMP and cGMP have several downstream targets including protein kinases that phosphorylate other proteins and regulate various cellular processes.
Glutamate and glycine are important neurotransmitters in the central nervous system. Glutamate is the primary excitatory neurotransmitter and is synthesized from glucose or glutamine. It acts through ionotropic and metabotropic glutamate receptors. The three main ionotropic receptor subtypes are NMDA, AMPA, and kainate receptors. NMDA receptors require both glutamate and glycine to open the ion channel. Metabotropic glutamate receptors are G protein-coupled and modulate intracellular signaling pathways. Glycine is also a neurotransmitter and is required as a co-agonist for NMDA receptor activation, playing a role in excitatory neurotransmission.
This document summarizes several anticonvulsant drugs, including their mechanisms of action and metabolic pathways. It discusses how primidone is metabolized to phenobarbital, how carbamazepine and oxcarbazepine are metabolized by cytochrome P450 enzymes, and how gabapentin and pregabalin are not metabolized. It also provides information on the metabolic pathways and side effects of other anticonvulsants such as lamotrigine, topiramate, zonisamide, levetiracetam, tiagabine, ethosuximide, and vigabatrin. The document concludes by mentioning the uses and metabolic pathways of clonazepam,
This document discusses glutamate receptors, including their history, types, roles, and drugs that act on them. It notes that glutamate is the major excitatory neurotransmitter in the central nervous system. There are two main types of glutamate receptors: ionotropic receptors which are ligand-gated ion channels including NMDA, AMPA, and kainate receptors, and metabotropic G protein-coupled receptors divided into groups 1, 2, and 3. The roles of glutamate receptors include synaptic plasticity, learning and memory, and excitotoxicity. Many drugs have been developed that act as agonists or antagonists at glutamate receptors and are being investigated for conditions like Alzheimer's, Parkinson
This document summarizes 5 major categories of transducer mechanisms:
1) G-protein coupled receptors which activate downstream effectors like adenylyl cyclase or phospholipase C.
2) Ion channel receptors which directly open or close ion channels.
3) Transmembrane enzyme-linked receptors which activate intracellular protein kinases.
4) Transmembrane JAK-STAT binding receptors which activate the JAK/STAT signaling pathway.
5) Receptors regulating gene expression which bind intracellularly to directly regulate gene transcription.
- GABA is the major inhibitory neurotransmitter in the mammalian brain. It acts through GABAA, GABAB, and GABAC receptors.
- GABAA receptors are ligand-gated chloride channels modulated by drugs like benzodiazepines, barbiturates, and general anesthetics. GABAB receptors are G-protein coupled receptors that inhibit neurotransmitter release and hyperpolarize neurons.
- Drugs that enhance GABAergic transmission through GABAA receptors like benzodiazepines are used as sedatives, anxiolytics, and anticonvulsants. The GABAB agonist baclofen is used as a muscle relaxant for spastic
1. G-proteins bind GTP and control intracellular signaling pathways. They exist in two states - active when bound to GTP and inactive when bound to GDP.
2. G-proteins are tightly regulated by accessory proteins that modulate their cycling between GTP-bound and GDP-bound states.
3. The heterotrimeric G-proteins transmit signals from cell surface receptors to enzymes and channels. They are stimulated by receptors, act on effectors, and are regulated by nucleotide exchange and hydrolysis.
This document summarizes information about the neurotransmitters glutamate and GABA. It discusses their roles, receptors, and involvement in various neurological conditions and disorders. Glutamate is the primary excitatory neurotransmitter in the brain, while GABA is the primary inhibitory neurotransmitter. The document outlines their synthesis, metabolism, receptor types, and relationships. It also explores how imbalances in glutamate and GABA are implicated in conditions like anxiety, depression, addiction, and neurodegenerative diseases.
G Protein Coupled Receptors (GPCRs) and CancerDayananda Salam
GPCRs play an important role in many types of cancer through their activation by various ligands. Some key points:
- GPCRs make up over 1% of the human genome and are the target of over 50% of drugs. They signal through G proteins and regulate many important cellular functions.
- In cancer, GPCRs activated by ligands like LPA, SDF1, chemokines promote processes like growth, migration, angiogenesis and metastasis. Examples include LPA1 in breast cancer and CXCR4 in ovarian cancer.
- GPCRs can also interact with other pathways like EGFR to enhance signaling. Orphan GPCRs without known ligands may play emerging roles in cancers like GPR49
Phenytoin is metabolized into reactive intermediates like arene oxide and catechol that can bind to proteins and trigger immune responses. These metabolites are deactivated by enzymes like epoxide hydrolase and glutathione transferase. Phenytoin strongly induces several CYP450 isozymes and drug metabolizing enzymes. Carbamazepine is metabolized by CYP3A4 into an epoxide metabolite suspected of causing idiosyncratic reactions. Gabapentin and pregabalin modulate calcium influx and stimulate GABA biosynthesis without being metabolized. Felbamate undergoes hydroxylation and hydrolysis to form a toxic metabolite, 2-phenylpropanal. Future anticonvulsants
GABA is the primary inhibitory neurotransmitter in the central nervous system. It acts on three main receptor types: GABAA, GABAB, and GABAC. GABAA is a ligand-gated ion channel whose activation allows chloride ion influx. GABAB is a G-protein coupled receptor whose activation opens potassium channels and closes calcium channels. Anti-epileptic drugs can act on these GABA receptors and neurotransmitter pathways. Newer anti-epileptics discussed include lamotrigine, gabapentin, topiramate, levetiracetam, zonisamide, tiagabine, and vigabatrin. Their mechanisms of action involve effects on sodium channels,
Dr. Pavani discusses G protein-coupled receptors (GPCRs), which are integral membrane proteins that sense molecules outside the cell and activate internal signal transduction pathways and cellular responses. There are over 800 GPCRs in humans that detect a wide range of ligands and are involved in many physiological processes. GPCRs work by coupling to G proteins, which activate various intracellular effectors like adenylyl cyclase, phospholipase C, and ion channels. Dysregulation of GPCR signaling can lead to many human diseases. Martin Rodbell and Alfred Gilman were awarded the 1994 Nobel Prize in Physiology or Medicine for their discoveries related to G proteins and GPCR signal transduction.
Assignment on Secondary messengers and intracellular signalingDeepak Kumar
Assignment on Secondary messengers: cyclic AMP, cyclic GMP, calcium ion, inositol 1,4,5- trisphosphate, (IP3), NO, and diacylglycerol. Detailed study of following intracellular signaling pathways: cyclic AMP signaling pathway, mitogen-activated protein kinase (MAPK) signaling, Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling pathway.
The document discusses neurotransmission and non-adrenergic, non-cholinergic neurotransmission. It describes how neurotransmitters are synthesized, packaged into vesicles, released into the synaptic cleft upon neuronal stimulation, and bind to receptors on the postsynaptic neuron. It notes that some neurons release multiple neurotransmitters, including glutamate, ATP, nitric oxide, and peptides. The presentation focuses on glutamate as a major excitatory neurotransmitter in the central nervous system that acts through ionotropic AMPA, kainate, and NMDA receptors and metabotropic receptors. Disorders associated with glutamate dysregulation and its role in memory and learning are also mentioned.
The document summarizes G protein-coupled receptor (GPCR) signaling. It discusses how different GPCRs are coupled to different G proteins that activate various downstream effectors like adenylyl cyclase, phospholipase C or ion channels. It provides examples of specific GPCRs like beta-adrenergic receptors and muscarinic acetylcholine receptor and their signaling mechanisms. It also describes the role of second messengers like cAMP, IP3 and DAG in GPCR signaling.
This document discusses a study examining the interaction between dopamine and histamine in the basal ganglia. It provides background information on synaptic transmission, the histaminergic system, dopamine signaling in the striatum, and Parkinson's disease. The study aims to investigate how activation of dopamine D1 receptors regulates phosphorylation of glutamate AMPA receptors and DARPP-32 in striatal slices, and how histamine H3 receptor activation may oppose these effects of dopamine signaling. The results suggest dopamine increases phosphorylation of AMPA receptors and DARPP-32, while histamine H3 receptor activation reduces dopamine-induced phosphorylation, indicating opposing regulation of cAMP/PKA signaling by dopamine and histamine in striatal neurons.
G protein-coupled receptors (GPCRs) are seven transmembrane receptors that activate intracellular signaling pathways through heterotrimeric G proteins. When an agonist binds to a GPCR, it causes a conformational change activating the G protein's α subunit. The activated α subunit then interacts with downstream effectors like adenylyl cyclase or phospholipase C. This leads to the production of second messengers like cAMP or IP3. Mutations in GPCRs and G proteins can cause disease by disrupting signaling, with loss or gain of function mutations associated with various endocrine disorders and cancers.
This document compares two methods for sequencing proteins: Edman degradation and mass spectrometry. Edman degradation involves tagging and removing amino acids from the N-terminal end one by one to determine the sequence. Mass spectrometry involves digesting proteins into peptides, using mass to charge ratios to separate peptides, and fragmenting peptides to measure fragment ion masses and deduce sequences. Mass spectrometry is more sensitive, does not require purified samples, and can handle modified proteins better than Edman degradation.
Alcohol related changes in the regulation of NMDA receptor functions-József NagyNiyamat Chimthanawala
A concise review about all the events supposedly playing a
regulatory role in the up-regulation of NMDAR functions in
consequence of chronic ethanol exposure.
The effect of subunit expression, molecular mechanisms, protein kinase/phosphatases activity, post-synaptic density, allosteric modulators activity is observed.
G Protein–Coupled Receptors (GPCRs) are integral membrane proteins that are activated by extracellular signaling molecules and activate intracellular secondary messenger pathways. They have seven transmembrane domains and activate heterotrimeric G proteins upon ligand binding. The G protein then activates downstream effector enzymes like adenylyl cyclase, which generates secondary messengers like cAMP. These messengers go on to activate pathways that ultimately alter cell function. The beta-adrenergic receptor pathway is a key example, with epinephrine binding and cAMP production leading to protein kinase A activation. PKA then phosphorylates target proteins to produce effects like increased heart rate. GPCRs are major drug targets, and their deregulation
Neurohumoral transmission in CNS ,special emphasis on importance of various neurotransmitters like with GABA, Glutamate, Glycine, serotonin and dopamine
The document discusses second messenger systems. It describes how second messengers relay signals from cell surface receptors to target molecules inside the cell. Some key points discussed include:
- Earl Sutherland discovered cyclic AMP (cAMP) as the second messenger for epinephrine and won the Nobel Prize for this work.
- Common second messenger systems include those using cAMP, cGMP, phosphatidylinositol, and tyrosine kinases as secondary messengers.
- G proteins act as transducers between receptors and effectors and are important drug targets.
- cAMP and cGMP have several downstream targets including protein kinases that phosphorylate other proteins and regulate various cellular processes.
This document discusses the role of AMPA receptor (AMPAR) surface diffusion in synaptic plasticity and memory. It reports that:
1) AMPAR surface diffusion is important for the establishment and maintenance of long-term potentiation (LTP) through replenishing synaptic AMPARs.
2) Blocking AMPAR surface diffusion through crosslinking attenuates LTP in hippocampal brain slices and impairs hippocampal-dependent memory formation.
3) Postsynaptic AMPAR surface diffusion is a critical trafficking mechanism for the expression of LTP and learning in the hippocampus.
The document discusses adrenergic drugs and their mechanisms of action. It notes that adrenergic drugs act on adrenergic receptors located presynaptically or postsynaptically. They affect the heart rate and contractility, blood vessel resistance, release of insulin, and lipolysis. The document then details the processes of neurotransmission at adrenergic neurons including synthesis, storage, release, binding and removal of neurotransmitters like norepinephrine. It describes the different types of adrenergic receptors and their subtypes, and the mechanisms of action and effects mediated by stimulating each receptor subtype.
This document summarizes research evaluating a drug for the treatment of Parkinson's disease. It describes:
1) In vitro and in vivo models used to test the drug, including studies using primary microglial cultures, rat striatal slices, and assays of dopamine stimulated adenylyl cyclase activity.
2) In vivo behavioral models to evaluate the drug's efficacy in treating Parkinsonism symptoms, such as reserpine antagonism tests, neuroleptic induced parkinsonism models, and skilled paw reaching tests.
3) The drug was evaluated using various in vitro and in vivo models to assess its ability to reduce inflammation, protect dopamine neurons from oxidative stress, and alleviate motor symptoms of Parkinson's disease
Glutamate is the major excitatory neurotransmitter in the central nervous system. It acts through ionotropic and metabotropic receptors. Ionotropic receptors are ligand-gated ion channels composed of NMDA, AMPA, and kainate receptor subtypes that allow cation influx. Metabotropic receptors are G protein-coupled receptors divided into three groups based on sequence homology and signaling pathways. Glutamate receptors play important roles in normal neurotransmission but excessive activation can lead to excitotoxicity involved in various neurological disorders.
This document summarizes G protein-coupled receptors (GPCRs) and ligand-gated ion channels. It notes that GPCRs are the largest family of receptors, with seven transmembrane domains that activate G proteins to trigger intracellular signaling pathways. Ligand-gated ion channels directly form ion channels when activated by neurotransmitters like acetylcholine and GABA. Second messengers downstream of GPCR signaling like cAMP and phosphoinositides are also discussed.
This document provides information about a receptor pharmacology course taught by Professor Dr. Md. Shah Amran at the University of Dhaka. It was prepared by 5 students and contains contents on different types of receptors including ligand gated ion channels, G-protein coupled receptors, enzyme linked receptors, nuclear receptors, and a comparison of receptor types. Receptors are important macromolecules that bind drugs and mediate their effects in the body.
This document provides information about a receptor pharmacology course taught by Professor Dr. Md. Shah Amran at the University of Dhaka. It was prepared by 5 students and contains an introduction to receptors, classifications of different receptor types including ligand gated ion channels, G-protein coupled receptors, enzyme linked receptors, and nuclear receptors. It also discusses receptor-drug interactions, affinity, intrinsic activity, and mechanisms of cell surface and intracellular receptors.
This document provides an overview of neurotransmitters and receptors. It discusses the mechanisms of fast neurotransmission mediated by neurotransmitters directly activating ligand-gated ion channels, as well as neuromodulation mediated by neurotransmitters binding to G-protein coupled receptors. The document outlines the major neurotransmitters - glutamate, GABA, glycine, acetylcholine - and covers their synthesis, release, reuptake, degradation and receptor systems. It also touches on the pharmacology of receptor agonists and antagonists.
Participation of the gabaergic system on the glutamateankit
This study investigated the participation of the GABAergic system in regulating glutamate release from frontal cortex synaptosomes in a rat model of experimental autoimmune encephalomyelitis (EAE), which mimics multiple sclerosis in humans. The results showed reduced GABAergic inhibition of glutamate release and impaired GABA regulation of synapsin I phosphorylation in synaptosomes from EAE rats compared to controls. Specifically, GABA had no effect on glutamate release or synapsin I phosphorylation in EAE rats, suggesting alterations in the GABAergic system may contribute to cortical pathology in EAE.
The document discusses the role of glutamate neurotransmission in various psychiatric and substance use disorders. It notes that a paradigm shift from focusing on monoamine hypotheses to a neuroplasticity hypothesis centered around glutamate may help advance treatment for depression. It also discusses how thinking about schizophrenia and addiction from a glutamate perspective provides new receptor targets and conceptual opportunities. The document provides details on glutamate regulation and interactions with dopamine systems. It reviews evidence that drugs of abuse interact with and cause long-lasting changes to glutamate transmission. Two case reports are presented showing lamotrigine's effectiveness in reducing substance craving and symptoms by potentially impacting glutamate neurotransmission.
My presentation on neurotransmitter glutamate. References from Comprehensive textbook of psychiatry 9th edition and Stahl's essential psychopharmacology 4th edition.
Agonists at adrenergic receptors are either direct-acting or indirect-acting. Catecholamines, norepinephrine, and epinephrine are direct-acting and nonselective adrenergic agonists. Indirect-acting agonists cause the release of the neurotransmitter norepinephrine from sympathetic nerve terminal
Cyclic adenosine monophosphate (cAMP) is an important second messenger in intracellular signal transduction. It is derived from ATP and conveys signals from hormones that bind to cell surface receptors. Many hormones such as epinephrine, glucagon, and others activate adenylate cyclase, which produces cAMP from ATP. cAMP then activates protein kinase A and triggers physiological responses in the cell. The effects of cAMP are terminated by phosphodiesterases that break it down or by phosphatases dephosphorylating protein kinase A. Deregulation of cAMP pathways has been implicated in cancer and cognitive disorders.
G protein coupled receptor and pharmacotherapeuticspriyanka527
This document provides an overview of G-protein coupled receptors (GPCRs) and their role in cell signaling. It discusses the history and structure of GPCRs, how they interact with G-proteins and secondary messengers like cAMP and IP3 to activate intracellular signaling pathways. These pathways regulate key cellular processes and are targets for drug development to treat diseases. The document also categorizes different classes of GPCRs and summarizes the mechanisms and physiological roles of various secondary messenger systems like cAMP, IP3, and ion channels in signal transduction.
GABA, glutamate receptors and their modulationDrSahilKumar
This document provides an overview of glutamate and GABA, their receptors and therapeutic applications. It discusses the synthesis, storage, release and termination of glutamate and GABA in the central nervous system. It describes the ionotropic and metabotropic glutamate receptors and GABAA and GABAB receptors. It also discusses conditions associated with glutamate like seizures, neurodegenerative diseases and stroke. Finally, it outlines current and upcoming therapeutic agents that target glutamate and GABA receptors and their uses, mechanisms and adverse effects.
G protein-coupled receptors (GPCRs) are integral membrane proteins that detect extracellular molecules and signal intracellular pathways. They have seven transmembrane domains and interact with G proteins. There are two principal signaling pathways: 1) the cAMP pathway, where the G protein activates adenylyl cyclase to produce cAMP, which activates protein kinase A and regulates genes and secretion; and 2) the phosphatidylinositol pathway, where the G protein activates phospholipase C to produce IP3 and DAG, which trigger calcium release and activate protein kinase C to phosphorylate target proteins and induce cellular responses. These pathways are connected by calcium-calmodulin, which regulates enzymes in both routes.
1. Second messengers are small intracellular molecules that transmit signals within cells after extracellular signaling molecules (hormones or neurotransmitters) bind to cell surface receptors.
2. There are three main types of second messenger systems: cyclic AMP (cAMP), cyclic GMP (cGMP), and inositol trisphosphate (IP3)/diacylglycerol (DAG). These systems activate protein kinases or trigger the release of calcium ions to produce a physiological response.
3. Second messengers amplify and diversify extracellular signals, allowing for precise regulation of multiple cellular processes. Their roles are important for understanding cell signaling, disease mechanisms, and potential drug targets.
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3. Introduction
● L-glutamate is the major excitatory
neurotransmitter in the mammalian
CNS
● It’s implicated in fundamental brain
functions, such as neuronal
development, learning and memory
3
5. Topology of AMPA Receptors
• AMPA Receptors are responsible for the majority of
fast excitatory synaptic transmission
• Recent findings have implicated AMPA Receptors in
synapse formation and stabilization; (Huganir and
Nicoll 2013)
• AMPA Receptor overactivation is potently
excitotoxic and changes in AMPA Receptor activity
have been described in the pathology of numerous
diseases
5
(Schiffer and Heinemann 1999)
Single Receptor
9. Objectives
• Assaying and identifying potent curcumin derivatives that are selective towards
AMPA Receptors only
• Characterizing the AMPA Receptor through the inhibition effect of curcumin
derivatives
• Promoting the creation of small interfering or mimicking peptides for up or down
regulation of AMPA Receptor activity at the synapse
• Identifying the functional groups accountable for the inhibition effect on AMPA
Receptor
9
11. Significance
One of the most apparent significances for characterization of the action
of Curcumin derivatives on the AMPA receptors is to find the most
potent and selective natural source of AMPA receptors inhibitors and to
use these findings for the purpose of synthesizing newer natural
effective drugs without the chemical side effects
11
12. Specific Aims
• Transfecting GluA2 receptor into Human Embryonic Kidney 293 (HEK293) cells
successfully.
• Characterizing the curcumin derivative A effect on AMPA receptor whole cell
current
• Characterizing the curcumin derivative A effect on AMPA receptor
desensitization
• Characterizing the curcumin derivative A effect on AMPA receptor deactivation
12
20. What can we conclude from this session?
● Curcumin derivative A decreased the peak amplitude of whole cell current, increased
desensitisation rate and extent and increased deactivation rate
● Curcumin derivative A is an inhibitor of AMPA receptor
● Curcumin derivative A might have affected the density of AMPA Receptors in the post
synaptic cleft by altering the trafficking mechanisms of AMPA Receptors
● Curcumin derivative A might have modified the chemical properties of the AMPA
Receptors channel pore, making it less permeable to certain ions
● Curcumin derivative A might have an effect on the conformation of the AMPA
Receptors, destabilizing the desensitization state and reducing the period of time
AMPA Receptors spends in that state
20
21. What can we conclude from this session?
● The modified curcumin derivative A contained an electron donating group which we
suggest plays a key a role in the inhibition effect of the curcumin,it might enhance
the ability of the compound to bind tightly to the AMPA Receptor
● The natural product; Curcumin and its derivatives are a target worth pursuing for
further research
21
22. Acknowledgment
Advisor
Dr. Mohammed Qneibi
Lab Technician:
Mrs. Shuruq Subuh
Chemistry Assistant Professor:
Dr. Othman Hamed
Examination Committee Members
22
This work was supported by MoHE grant.
Good morning everyone. It’s my pleasure to present to you our research today, my name is Hasan Arafat, my colleagues Yasmeen Abu Naba and Remah Yousef, we will be presenting The Effect of Curcumin on AMPA Receptor Kinetics.
So before I start, let me outline what we’re going to talk about in this session. We’ll start off by giving a brief background, the objective and significance of our study, followed by the specific aims and how we conducted this study.our results and Finally a summary and what our research will provide for future studies.
So let me start with the introduction. L-glutamate is the major excitatory neurotransmitter in the mammalian CNS. It’s the most abundant neurotransmitter in the vertebrate CNS. L-Glu is implicated in fundamental brain functions, such as neuronal development, learning and memory. So now, let’s talk briefly about the mechanism by which glutamate conveys its actions.
As action potential reaches the axon terminus, the pre-synaptic membrane depolarizes, opening voltage gated Ca2+, prompting an influx of Ca2+.
Glutamate vesicles fuse with the membrane, releasing glutamate into the cleft.
Glutamate diffuses from the presynaptic membrane through the synaptic cleft in a fraction of millisecond, and binds glutamate receptors at the post synaptic cleft.
This binding changes the permeability of the receptor, ultimately generating a membrane potential at the post-synaptic membrane.
And now let’s talk about glutamate receptors. Glutamatergic receptors can be divided into metabotropic glutamate receptors, a.k.a G-protein coupled receptors, and ionotropic glutamate receptors. For the sack of our discussion, we will focus on ionotropic receptors. Ionotropic receptors are further classified into Kainate, AMPA, and NMDA receptors.
1- Kainate receptors control synaptic integration and spike transmission
2- AMPA receptors mediate fast excitation
3- NMDA receptor activity correlates to a much slower and long-lasting current profile
Each type has its own set of subunits, we will focus on AMPA receptors.
AMPA receptors are composed of four receptor subunits: GluA1 to 4. AMPA receptors can form
homomeric functional channels and heteromeric channel complexes.
GluA1-GluA2 (most abundant in hippocampus) and GluA2-GluA3 combinations make the majority of AMPA Receptors there. On the other hand, GluA4-containing AMPA Receptors are expressed mainly in early postnatal development.
GluA1 forms calcium-permeable (without GluA2), and is associated with LTP, and LTD, which affect the synaptic plasticity.
GluA1 functions in the trafficking and insertion of AMPA Receptors in the synapses, hence its unique role in various nervous disorders
GluA2 has the most impact on the biophysical properties of all heteromeric complexes. GluA2-lacking AMPA Receptors are Ca2+ permeable. In contrast, GluA2-containing AMPA Receptors are impermeable to divalent cations, particularly Ca2+. GluA2 receptors also have a charechterisitc form of kinetics
They are also implicated in receptor plasticity, which is defined as the ability of the brain to change and adapt to new information; Synaptic plasticity is a biological process in which synaptic strength changes in retaliation to a specific pattern of changes in neuronal activity.
Point Two: each subunit is highlighted in different color in this scheme, (point using the flashlight), this is the amintotropic domain, ligand binding domain and the transmembrane domain
Homomeric: all subunits are identical (all GluA1, for example), hetermeric: different subunits.The Synaptic Strength is measured by the degree of changing occurring in the postsynaptic neuron potential in respect to changes in presynaptic stimulation.An essential contributor to the synaptic strength is the fact the AMPA receptors are not a fixed element in the synaptic composure, instead are characteristically dynamic, being moved in and out of a synapse in response to neuronal activity.LTP: Long Term Potentiation:LTD: Long Term Depression:
Trafficking their over-activation is potently excitotoxic triggering either rapid or delayed neurotoxicity. Changes in AMPA Receptor activity and consequently a debilitated regulation of Glutamate have been described in the pathology of several diseases, such as ALS, stroke, epilepsy and Alzheimer's disease.o
So as you can see in this figure, the ligand binding site is represented by the clamshell structure, with D1 representing the upper valve of the shell and D2 representing the lower valve. The gray bars represent the ion channel. In part A here, the receptor is in the resting state, there is no ligand at the LBD, and the ion channel is closed. The current passing through the channel is represented here as a flat trace. In part B, the ligand binds to the LBD, opening the ion channel and allowing ions to pass through. This is represented as an abrupt change in the current trace as you can see. The following step is known as desensitization, in which the ligand remains bound to the LBD, while the ion channel becomes loosely closed, decreasing the passage of ions but allowing a small amount to leak, this is represented as a gradual change in the current trace as you can see. The steady state here is considered a part of the desensitization phase, in this phase, the current reaches a plateau in which the electrical charge leaving the cell is equal to that entering the cell. Then, the receptor returns to the resting state, in which there is no ligand at the LBD, the ion channel is closed.
With this I have explained the 1 important phenomenon that our research has focused on, now since we understand that importance of AMPA Receptor and understand how their overactivation is linked to excitotoxicity and to CNS diseases , to stop this from happening we should explore compounds that stop this over excitation and neuronal death from happening ,so now we'll explain what we used for AMPA Receptor inhibition.
The compounds shown here are AMPA Receptor antagonist or inhibitors and are drugs used for many purposes. As I said, AMPA Receptors are excitotoxic and their overactivation is implicated in many diseases. All of these are chemical compounds that come with many systemic side effects, this matter aspired us to look for natural compounds that work as efficiently but don’t come with the annoying side effects.
This has lead us to curcumin >>> next silde
This lead us to Curcumin, is a well known spice with a scientific history going back two centuries ago when it was first isolated from Turmeric (Curcuma longa) roots.
It’s known for its anti-oxidant , anti-inflammatory, chemotherapeutic and chemopreventive effect.
It’s also known for its neuroprotective effect, esp. Alzheimer disease, ALS and epilepsy.
It’s a potent Reactive Oxygen Species (ROS) scavenger, at least ten times more potent than Vit. C.
Now this compound shown above is native curcumin, the compound shown below is the derivative we chose for our study. Due to time limitation, we chose to study this one compound although we had a variety of other compound, which we will study is the future.
And here we finish with the introduction, we talked about glutamate, glutamate receptors and their types, AMPA receptor, AMPA subunits and kinetics and curcumin. Now I leave you with my colleague Yasmeen Abu Nabaa to continue with the objectives, Yasmeen.
The reason why we chose this particular derivative is because of the amine group it contains, which is an electron donating group as you know. According to a previous study of Dr. Qneibi, BZD, the well-known CNS inhibitors, inhibit AMPA Receptors peak current, this was found to be due to the interaction of the electron-donating group of BDZ with AMPA Receptors. We suggested a similar role for this derivative based on the amine group it contains, and this is why we chose to test it first.
Limitation: We did not have enough time to study all components ..
** to understand the molecular and cellular mechanisms underlying synapse
development and function.
** but due to the limitations ( opening of lab only recently and deadline) we did not have enough time to do this so we started with 1 but will continue on this objective for our future study.
Now this table shows the different derivatives we obtained from curcumin, all of these compounds will be used in our future studies.
As we previously mentioned, AMPA Receptor has been implicated in many CNS diseases , this makes it a target worth pursuing.
Extraction was done by Dr Othman in faculty of chemistry. Curcuma longa powder was placed in a Soxhlet extractor and extracted with methanol for about 4 hr. The methanol solvent was filtered then concentrated under vacuum. The yellow gummy residue containing curcuminoids was subjected to purification by flash chromatography on silica (100-200 mesh).first fraction was eluted with hexane-ethyl acetate and then methanol-ethyl acetate was used to elute the second fraction. It was found that the Second fraction contains the desired curcumin.
DNA preparation:
Cell culture (Human Embryonic Kidney 293) cells were grown in supplemented DMEM (Dulbecco Modified Eagle Medium) supplemented with 10% FBS (fetal bovine serum), 0.1 mg/ml streptomycin, and 1 mM sodium pyruvate.
the picture here shows healthy grown HEK293 under microscope.
Transfection the process of deliberately introducing naked or purified nucleic acids into eukaryotic cells. (plasmid containing AMPA Receptor gene into HEK293 cells in our case) .Before the electrophysiology recordings, cells were replated 24 hours after transfection on coverslips coated with Laminin.
The right image here shows transfected HEK293 under fluorescent microscope, the green light represents the fluorescent dye eGFP (enhanced Green Fluorescent Protein) found on the plasmid carrying the gene for the AMPA Receptor. These shiny large spots represent free dye, these irregular shapes are deformed cells that are unsuitable for patch-clamp, circular, well defined cells represent healthy cells that took the plasmid and has mostly expressed the receptor of our interest. All of these cells (point using the laser) represent potential candidates for patch-clamp.
Wash System: made of 5 syringes, the first one contains the wash solution, the second contains glutamate, and the other three contains distilled water.
Glutamate, l
Wash, a buffer solution
As you can see, we attach the pipette to the healthiest cell in the field. We used the whole cell patch clamp, in which only a small part of the membrane is ruptured using the pipette, this allows direct recording of the cell current. The solution flowing through the pipette is known as the internal solution, and it’s carefully prepared to simulate the cytoplasm. As the pipette attaches to the membrane, a GigaOhm seal is applied at the membrane. The whole cell is carried using the pipette outside the field and exposed to the external solution barrel of the theta, this keeps the cell at base current and viable. At command, the theta sweeps to the glutamate barrel in 20 μs second, exposing the cell to glutamate and activating AMPA Receptors, generating an action potential.
At this step, the wash, a.k.a the external solution, a solution prepared to simulate that of the external environment surrounding the cell, is applied to the cell using one of the theta two barrels. When the user is ready for the experiment, glutamate is applied using the other barrel, eliciting a change in the membrane action potential due to the binding of the glutamate to the LBD at AMPA Receptors, the current elicited is measured using the pipette
simultaneously, the wash solution starts flowing through the theta, the wash system contains certain chemicals that keep the cell at base current and keep it healthy and alive while a part
We first studied the effect of curcumin derivative A on glutamate-evoked AMPA receptor-mediated whole cell currents in HEK293 Cells.
The amplitude generated by GluA2Q receptors was measured using Integrated Patch Amplifiers (IPA),
Agonist was applied on GluR2Q by using Piezo Fast Exchange solution with 10 mM of glutamate.
Data were analyzed using Igor 7 software.
As shown in figure A, application of Curcumin derivative A in agonist barrels showed significant decrease in the peak glutamate-activated current of GluA2Q.
The amount of peak current has decreased from 4820 pA to 3012 pA (i.e. ~1.6 folds).
The y-axis of figure A was plotted as A/A1, which is a function of inhibitor concentration vs. the concentration of the curcumin derivatives in the x-axis.
From the observed shortening of the activation limb of the AMPA receptor current trace resonating with the decrease in the peak amplitude. This suggest that the curcumin derivative may affect the activation phase time.????
We evaluated the effect of the derivative on desensitization. Following the activation of GluA2, superimposed current traces evoked by 500 ms pulses of 10 mm glutamate in AMPA Receptor alone and AMPA Receptor plus the curcumin derivative were obtained to study its effect on desensitization.
To recall , Desensitization is defined as the characteristic decay in current after a long simulation time following activation. Thus evoking for the period of 500ms allow us to observe this phenomenon and the slow recovery of the receptor.
curcumin derivative affected the desensitization which was markedly reduced following the administration of the curcumin as in figure B.
Comparison of desensitization time constants between AMPA Receptor alone (9.84 ± 1.64 ms, n = 4) and with the addition of curcumin (4.6 ± 1.1 ms, n = 4) shows a significant difference in the desensitization time constant.
The amount of desensitization can be measured by : peak/steady state
the rate of desensitization equals the inverse of time constant obtained by exponential fit to the desensitization (tau) using equation : 1/τ(s-1)
The amount of desensitization increased from an average of 19.08 to that of 30.78 ~1.6 folds, while the average rate of desensitization increased from 0.10 ms-1 to 0.22 ms-1 ~2 folds.
We focused our attention on deactivation.
Following the activation of GluA2, superimposed current traces evoked by 1 ms pulses of 10 mm glutamate in AMPA Receptor alone and AMPA Receptor with curcumin derivative were obtained to study the curcumin derivatives effect on deactivation.
To recall , Deactivation is the characteristic decay in current after a short simulation time following activation. Applying agonist for 1 ms allow us to catch the receptor rapid recovery upon the immediate removal of glutamate.
As shown in figure B the presence of curcumin derivative also affected deactivation, by a distinct decrease of the current peak, which increased the rate of receptor deactivation.
Comparison of deactivation time constants between AMPA Receptor alone (5.88 ± 0.40 ms, n = 5) and with curcumin (4.8 ± 0.6 ms, n = 5) variants (figure A) shows a notable difference in the deactivation time constant.
The decrease in time constant suggest that the curcumin derivative accelerated the rate of deactivation of GluA2.
The amount of deactivation, calculated using :peak/steady state,increased from an average of 895.37 to that of 977.25 ~1.1folds
calculated using equation 1/τ(s-1) : the average rate of deactivation increased from 0.17 ms-1 to 0.2 ms-1 ~1.18 folds
In the presence of curcumin, the time the receptor spent in the desensitized and deactivated phases was reduced, demonstrated by a reduction in the value of tau. This suggests that curcumin might have an effect on the AMPA receptor conformation, destabilizing the desensitized/deactivated conformation and favoring the resting conformation.
Point 2 ; Denoting that curcumin derivative renders AMPA Receptor less sensitive for its agonist glutamate.
Before point 3: We suggest these mechanism for the curcumin effect on AMPA Receptors.
وفي النهاية ،، نتقدم بجزيل الشكر وعظيم الامتنان الأهل الاعزاء الذين كانو مصدر القوة والالهام لنا ،، (وبنتمنى انه نكون رفعنا راسكم وفرحناكم بهاليوم ) وشكرا لجميع الأصدقاء والزملاء الذين كانو بجانبنا منذ بداية الطريق وشكرا لجميع الحضور والمستمعين ،،