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Making Optical and
Electrophysiological Measurements
in the Brain of Head-Fixed, Freely-
Moving Rodents
Sponsored by:
Making Optical and Electrophysiological Measurements in
the Brain of Head-Fixed, Freely-Moving Rodents
1. A brief history of The Mobile HomeCage (L. Khiroug, PhD)
2. Awake Plasticity (E. Castrén, MD, PhD)
3. Effects of drugs of abuse on dendritic spine plasticity:
intravitial microscopy in awake mice (P. Hyytiä, PhD)
4. Electrophysiological recordings of cortical activity in head-restrained
rodents in vivo (R. Khazipov, MD, PhD & M. Minlebaev, MD. PhD)
Sponsored by:
InsideScientific is an online educational environment
designed for life science researchers. Our goal is to aid in
the sharing and distribution of scientific information
regarding innovative technologies, protocols, research
tools and laboratory services.
A brief history of
The Mobile HomeCage
Leonard Khiroug
Chief Scientific Officer (CSO),
Neurotar Oy Ltd
• 10 years as a PI at Univ.
Helsinki (after postdoc at
Duke and NIH); Adj. prof
• In vivo two-photon
imaging as a service to
global Pharma industry
• Going awake: solving our
own problem => Product
for labs world-wide
Evolution to
Revolution
Academic
group
(2003-2013)
Service
business
(2009 - )
Device
business
(2014 - )
• Trans-BBB
Pharmacokinetics and
Biodistribution
Example:
Client’s proprietary antibody tagged
with Alexa Fluor 647 (red), kinetics
of penetration through BBB,
colocalization with Abeta tangles
labelled with Methoxy X04 (green)
In vivo microscopy
as a service
• Mitochondrial
fragmentation in Ischemic
Stroke models
Example:
Transgenic “reporter” mice expressing
mitoCFP on Thy1 promoter, visualizing
mitochondrial of cortical neurons.
Laser-induced stroke, reversible and
drug-targetable fragmentation of
mitochondria
In vivo microscopy
as a service
• Mitochondrial
fragmentation in Ischemic
Stroke models
Example:
Transgenic “reporter” mice expressing
mitoCFP on Thy1 promoter, visualizing
mitochondrial of cortical neurons.
Laser-induced stroke, reversible and
drug-targetable fragmentation of
mitochondria
In vivo microscopy
as a service
Note the network-like organization of elongated
mitochondria prior to the stroke induction
• Mitochondrial
fragmentation in Ischemic
Stroke models
Example:
Transgenic “reporter” mice expressing
mitoCFP on Thy1 promoter, visualizing
mitochondrial of cortical neurons.
Laser-induced stroke, reversible and
drug-targetable fragmentation of
mitochondria
In vivo microscopy
as a service
Note the fragmented mitochondria 30 minutes
after stroke induction (blood flow obstructed)
Full-body constraint
Previous solutions for studies in awake mice
Linear treadmill Head-mounted
devices
Air-lifted ball (spherical treadmill)
Previous solutions for studies in awake mice
Our Solution: “Flat Ball With a Wall”
Front view Side view Air
Natural enclosed environment, short habituation, stable fixation, compact design
Our Solution: “Flat Ball With a Wall”
Our Solution: “Flat Ball With a Wall”
 Two-photon
microscopy
 Optogenetics
 Intrinsic
optical
imaging
 Glutamate
uncaging
 Voltage-
sensitive dyes
 Whole-cell
patch clamp
 Multichannel
electrodes
 Voltammetry
 Ion-sensitive
electrodes
 Microdialysis
Sleep research in
head-fixed mice
Chronic recording (wire electrodes)
Acute recording (multi-shank high-density silicon probes)
Locomotion
Sleep
Run
Dr. Nikolaos Karalis and
Prof. Anton Sirota
(LMU, Munich, Germany)
Sleep research in
head-fixed mice
Dr. Nikolaos Karalis and
Prof. Anton Sirota
(LMU, Munich, Germany)
Sleep
Run
Sleep research in head-fixed mice
Head-fixed
in Mobile
HomeCage
Head-fixed
on treadmills
(linear, spherical)
+ VR
Carrying
head-mounted
miniaturized
devices
Short habituation and training,
minimized stress
+ - -
Cost-efficiency + - -
Natural environment
(flat floor, walls, obstacles)
+ - +
Allows high quality optics, multiple
precision electrodes
+ + -
Compactness and compatibility with
other equipment
+ - +
Allows place cell research or locomotion
over long distances
- + +
Summary
Awake Plasticity
Eero Castrén, MD, PhD
Neuroscience Center
University of Helsinki
Finland
Neuronal Plasticity:
• The capacity of neurons and neural circuits to change structurally and
functionally in response to experience
• Plasticity is regulated by neuronal activity
• Plasticity is bidirectional: adding and removing
• Neurogenesis => selection by programmed cell death
• Axon growth => selection by retraction
• Synaptogenesis => selection by synaptic pruning
• Synaptic potentiation and depression
• Active neurons and synapses are selected and stabilized
• NOT THE QUANTITY, BUT THE QUALITY
CTRL ANESTH
CTRL ANESTH
Question &
Experimental
Procedure
AN
30’
M M MM
24 h 1h 1h 24 h
Does anesthesia
influence dendritic
spine plasticity?
• Steady state spine
number or shape?
• Spine dynamics?
C A C A C A
Generally about
technique
• 2-photon imaging in
awake, freely
moving mice
Generally about
technique
• Allows repeated
imaging of an
identified neuronal
process
Isoflurane does not influence spine dynamics
Somatosensory cortex
• Imaging, stimulation and electrophysiology in living brain
• Increasingly evidence that anesthesia influences
• Need for recording/ imaging in awake animals
• Imaging/recording in behaving and learning animals
• Behavior in familiar, non-stressed environment
• Imaging in home cage environment would be optimal
ERA of connectomics
Effects of drugs of abuse on
dendritic spine plasticity:
intravitial microscopy in awake mice
Petri Hyytiä
PhD, adjunct professor,
Department of Pharmacology
Biomedicum
University of Helsinki Finland
• Introduction
• Why intravitial microscopy in awake mice?
• Our hypotheses
• Experimental design
• Preliminary data
• Summary of main points
What we are going to cover today?
The “Addicted” Spine
• Drug addiction marked by long-
lasting changes in behavior
• Persistent structural changes in
neurons in limbic brain regions
• Changes in size of cell bodies,
dendritic arborization, spine
morphology and dynamics
The brain reward pathways
Spiga et al. 2014
The “Addicted” Spine
• Drug addiction marked by long-
lasting changes in behavior
• Persistent structural changes in
neurons in limbic brain regions
• Changes in size of cell bodies,
dendritic arborization, spine
morphology and dynamics
• Robinson & Kolb 1997
• Medium spiny neurons in
nucleus accumbens
• First demonstration of drug-
induced structural plasticity
• Persistent increase in total
spines per 10 µm of dendrite
and in the number of spines
with multiple heads
(branched spines)
Amphetamine-
induced structural
modifications
Cocaine and morphine produce opposite effects in spine
density and dendritic branching
Cocaine Morphine
Fraction of spines gained
between imaging sessions
Cocaine induced structural plasticity in frontal cortex –
2-photon imaging via cranial windows
Spine accumulation during
cocaine treatment
New persistent spines gained
during cocaine conditioning:
correlation w/ cocaine CPP
Munoz-Cuevas et al 2013
Why intravitial microscopy in awake mice?
Benefits:
• Long-term longitudinal imaging
• No interfering anesthesia
• No need for control group
Limitation:
• Depth: only superficial layers of neocortex
• Other cortical areas:
motor, perirhinal,
orbital cortex
• Thalamocortical
projections
• Corticostriatal
projections
Primary
somatosensory
cortex (S1)
connectivity
Primary
somatosensory
cortex activation
by drugs of abuse
• Metabolic mapping
• Activation marker
c-Fos expression
• fMRI
• Changes in spine dynamics correlate with the
synaptic activity at the spines and therefore
serve as indicators of changes in neural circuitry
produced by drugs of abuse
• Persistence of the changes in spines point to a
mechanism behind the long-term drug effects,
including susceptibility to relapse
• Changes in dendritic spine turnover are
correlated with behavioral alterations
Our hypotheses
Thy1-YFP mice
Effects of subchronic morphine - Spine turnover imaging protocol
Week 1
M
1d
M M M
Week 3
M
1d
M
Week 2
MorphinePBS
Effects of subchronic morphine - Spine turnover imaging protocol
Week 1
M
1d
M M M
Week 3
M
1d
M
Week 2
MorphinePBS
Effects of subchronic morphine - Spine turnover imaging protocol
Week 1
M
1d
M M M
Week 3
M
1d
M
Week 2
MorphinePBS
Effects of subchronic morphine - Spine turnover imaging protocol
Week 1
M
1d
M M M
Week 3
M
1d
M
Week 2
MorphinePBS
Effects of subchronic morphine - Spine turnover imaging protocol
Week 1
M
1d
M M M
Week 3
M
1d
M
Week 2
MorphinePBS
Effects of subchronic morphine - Spine turnover imaging protocol
Week 1
M
1d
M M M
Week 3
M
1d
M
Week 2
MorphinePBS
Effects of subchronic morphine - Spine turnover imaging protocol
100 mm
Baseline, before PBS and Morphine 2 weeks after Morphine injections
100 mm
Imaging stability (raw data comparison)
• In vivo two-photon microscopy in non-anesthetized Thy1-
YFP mice yields unbiased information of drug-induced
changes in spine dynamics in the somatosensory cortex
• Monitoring these changes over extended periods of time
gives insight into drug-induced structural plasticity in the
mammalian neocortex
• Our pilot project with morphine demonstrates the
feasibility of the procedures using the mobile home cage
in the context of two-photon microscopy
Summary of main points
Electrophysiological recordings
of cortical activity in head-
restrained rodents in vivo
Roustem Khazipov
Directeur de Recherche,
INSERM U901
Marat Minlebaev
Charge de Recherche,
INSERM U901
1. Overview of the extracellular and patch-clamp
recording techniques (Khazipov)
2. Application of the Mobile HomeCage (Minlebaev)
3. Tips for stable electrophysiological recordings from the
head-fixed animals in Mobile HomeCage
What we are going to cover today:
Surface EEG
Intracortical recordings of the local field potentials
Multi-electrode arrays
Minlebaev & Khazipov, 2009
Intracortical recordings of the local field potentials
Minlebaev & Khazipov, 2009
• Local field potentials and
multiple unit activity (MUA -
spikes of individual neurons) can
be recorded from different
depth
• Current source density (CSD)
profile shows sinks and sources
of the population events
• MUA and cross-correlation
analysis shows how neurons in
different layers are activated
during population events
Multiple and single neuron action potentials
Mitrukhina et al., 2014
• Extracellular recordings
of neurons spikes with
tetrodes enable to
isolate spikes of
individual neurons
through cluster analysis
• One tetrode may give a
description of activity of
several neurons
Whole-cell recordings from a single neuron
Minlebaev et al., 2011
• Whole-cell recordings from
individual neurons enable
to access synaptic
correlates of the network
activity
• GABA and glutamate
synaptic currents can be
studied in isolation
• In current clamp mode,
firing of recorded cell and
subthreshold conductances
Cell-attached recordings
• Cell-attached recordings
enable to record spikes from
individual neurons and
• Currents through single ion
channels
• Reversal potential of GABA
currents can be deduced
from single GABA channel
activity without altering
intracellular chloride
Tyzio et al., 2006
1. Overview of the extracellular and patch-clamp
recording techniques (Khazipov)
2. Application of the Mobile HomeCage (Minlebaev)
3. Tips for stable electrophysiological recordings from the
head-fixed animals in Mobile HomeCage
What we are going to cover today:
Spontaneous hippocampal activity in CA1 region of awake mice
Theta oscillations
in Stratum
Radiatum
Cross-Frequency Coupling between Theta and Gamma
frequency bands in Stratum Radiatum
PAC measure = cfc Dataset = Structures = -
Ampfreq/Hz-
Phase freq / Hz -
5 10 15 20 25 30 35 40
0
10
20
30
40
50
60
70
80
90
0
0.005
0.01
0.015
0.02
0.025
0.03
0.035
0.04
0.045
0.05
Multi-unit activity (StrR and StrPyr) phase lock to StrRad
Local Field Potential
1s
Current clamp whole cell in vivo recordings
in the somatosensory cortex
2962029600295802956029540
Time (ms)
Signal00
(mV)
-90
-80
-70
-60
-50
-40
-30
-20
-10
0
3000029800296002940029200
Time (ms)
Signal00
(mV)
-90
-80
-70
-60
-50
-40
-30
-20
-10
0
• Coordinates (atlases, papers)
• Cortical surface better to keep wet (agarose better)
• Ensure mechanical stability (make the hole in the skull
as small as possible. Put dental cement close around
the hole)
• Minimize pulsations (put agar, silicone oil if needed)
• Avoid multiple insertions (not more than 5-10 attempts
through the same hole)
• Vertical penetration (‘+’ extra cortical immobilization,
‘-‘ tissue damage; ~10 penetrations).
• Pipette length depends on the recorded structure
(shorter – better, otherwise extra capacitance; for
striatum, up to 3 mm in mice)
Patch-clamping in awake mice
Thank You!
For additional information on the Mobile HomeCage and
methods for using this device in awake rodents with optical
imaging or electrophysiological techniques please visit:
http://www.neurotar.com/
InsideScientific is an online educational environment
designed for life science researchers. Our goal is to aid in
the sharing and distribution of scientific information
regarding innovative technologies, protocols, research
tools and laboratory services.

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Making Optical and Electrophysiological Measurements in the Brain of Head-Fixed, Freely-Moving Rodents

  • 1. Making Optical and Electrophysiological Measurements in the Brain of Head-Fixed, Freely- Moving Rodents Sponsored by:
  • 2. Making Optical and Electrophysiological Measurements in the Brain of Head-Fixed, Freely-Moving Rodents 1. A brief history of The Mobile HomeCage (L. Khiroug, PhD) 2. Awake Plasticity (E. Castrén, MD, PhD) 3. Effects of drugs of abuse on dendritic spine plasticity: intravitial microscopy in awake mice (P. Hyytiä, PhD) 4. Electrophysiological recordings of cortical activity in head-restrained rodents in vivo (R. Khazipov, MD, PhD & M. Minlebaev, MD. PhD) Sponsored by:
  • 3. InsideScientific is an online educational environment designed for life science researchers. Our goal is to aid in the sharing and distribution of scientific information regarding innovative technologies, protocols, research tools and laboratory services.
  • 4. A brief history of The Mobile HomeCage Leonard Khiroug Chief Scientific Officer (CSO), Neurotar Oy Ltd
  • 5. • 10 years as a PI at Univ. Helsinki (after postdoc at Duke and NIH); Adj. prof • In vivo two-photon imaging as a service to global Pharma industry • Going awake: solving our own problem => Product for labs world-wide Evolution to Revolution Academic group (2003-2013) Service business (2009 - ) Device business (2014 - )
  • 6. • Trans-BBB Pharmacokinetics and Biodistribution Example: Client’s proprietary antibody tagged with Alexa Fluor 647 (red), kinetics of penetration through BBB, colocalization with Abeta tangles labelled with Methoxy X04 (green) In vivo microscopy as a service
  • 7. • Mitochondrial fragmentation in Ischemic Stroke models Example: Transgenic “reporter” mice expressing mitoCFP on Thy1 promoter, visualizing mitochondrial of cortical neurons. Laser-induced stroke, reversible and drug-targetable fragmentation of mitochondria In vivo microscopy as a service
  • 8. • Mitochondrial fragmentation in Ischemic Stroke models Example: Transgenic “reporter” mice expressing mitoCFP on Thy1 promoter, visualizing mitochondrial of cortical neurons. Laser-induced stroke, reversible and drug-targetable fragmentation of mitochondria In vivo microscopy as a service Note the network-like organization of elongated mitochondria prior to the stroke induction
  • 9. • Mitochondrial fragmentation in Ischemic Stroke models Example: Transgenic “reporter” mice expressing mitoCFP on Thy1 promoter, visualizing mitochondrial of cortical neurons. Laser-induced stroke, reversible and drug-targetable fragmentation of mitochondria In vivo microscopy as a service Note the fragmented mitochondria 30 minutes after stroke induction (blood flow obstructed)
  • 10. Full-body constraint Previous solutions for studies in awake mice Linear treadmill Head-mounted devices
  • 11. Air-lifted ball (spherical treadmill) Previous solutions for studies in awake mice
  • 12. Our Solution: “Flat Ball With a Wall” Front view Side view Air Natural enclosed environment, short habituation, stable fixation, compact design
  • 13. Our Solution: “Flat Ball With a Wall”
  • 14. Our Solution: “Flat Ball With a Wall”  Two-photon microscopy  Optogenetics  Intrinsic optical imaging  Glutamate uncaging  Voltage- sensitive dyes  Whole-cell patch clamp  Multichannel electrodes  Voltammetry  Ion-sensitive electrodes  Microdialysis
  • 15. Sleep research in head-fixed mice Chronic recording (wire electrodes) Acute recording (multi-shank high-density silicon probes) Locomotion Sleep Run Dr. Nikolaos Karalis and Prof. Anton Sirota (LMU, Munich, Germany)
  • 16. Sleep research in head-fixed mice Dr. Nikolaos Karalis and Prof. Anton Sirota (LMU, Munich, Germany) Sleep Run
  • 17. Sleep research in head-fixed mice
  • 18. Head-fixed in Mobile HomeCage Head-fixed on treadmills (linear, spherical) + VR Carrying head-mounted miniaturized devices Short habituation and training, minimized stress + - - Cost-efficiency + - - Natural environment (flat floor, walls, obstacles) + - + Allows high quality optics, multiple precision electrodes + + - Compactness and compatibility with other equipment + - + Allows place cell research or locomotion over long distances - + + Summary
  • 19. Awake Plasticity Eero Castrén, MD, PhD Neuroscience Center University of Helsinki Finland
  • 20. Neuronal Plasticity: • The capacity of neurons and neural circuits to change structurally and functionally in response to experience • Plasticity is regulated by neuronal activity • Plasticity is bidirectional: adding and removing • Neurogenesis => selection by programmed cell death • Axon growth => selection by retraction • Synaptogenesis => selection by synaptic pruning • Synaptic potentiation and depression • Active neurons and synapses are selected and stabilized • NOT THE QUANTITY, BUT THE QUALITY
  • 21. CTRL ANESTH CTRL ANESTH Question & Experimental Procedure AN 30’ M M MM 24 h 1h 1h 24 h Does anesthesia influence dendritic spine plasticity? • Steady state spine number or shape? • Spine dynamics? C A C A C A
  • 22. Generally about technique • 2-photon imaging in awake, freely moving mice
  • 23. Generally about technique • Allows repeated imaging of an identified neuronal process
  • 24. Isoflurane does not influence spine dynamics Somatosensory cortex
  • 25. • Imaging, stimulation and electrophysiology in living brain • Increasingly evidence that anesthesia influences • Need for recording/ imaging in awake animals • Imaging/recording in behaving and learning animals • Behavior in familiar, non-stressed environment • Imaging in home cage environment would be optimal ERA of connectomics
  • 26. Effects of drugs of abuse on dendritic spine plasticity: intravitial microscopy in awake mice Petri Hyytiä PhD, adjunct professor, Department of Pharmacology Biomedicum University of Helsinki Finland
  • 27. • Introduction • Why intravitial microscopy in awake mice? • Our hypotheses • Experimental design • Preliminary data • Summary of main points What we are going to cover today?
  • 28. The “Addicted” Spine • Drug addiction marked by long- lasting changes in behavior • Persistent structural changes in neurons in limbic brain regions • Changes in size of cell bodies, dendritic arborization, spine morphology and dynamics The brain reward pathways
  • 29. Spiga et al. 2014 The “Addicted” Spine • Drug addiction marked by long- lasting changes in behavior • Persistent structural changes in neurons in limbic brain regions • Changes in size of cell bodies, dendritic arborization, spine morphology and dynamics
  • 30. • Robinson & Kolb 1997 • Medium spiny neurons in nucleus accumbens • First demonstration of drug- induced structural plasticity • Persistent increase in total spines per 10 µm of dendrite and in the number of spines with multiple heads (branched spines) Amphetamine- induced structural modifications
  • 31. Cocaine and morphine produce opposite effects in spine density and dendritic branching Cocaine Morphine
  • 32. Fraction of spines gained between imaging sessions Cocaine induced structural plasticity in frontal cortex – 2-photon imaging via cranial windows Spine accumulation during cocaine treatment New persistent spines gained during cocaine conditioning: correlation w/ cocaine CPP Munoz-Cuevas et al 2013
  • 33. Why intravitial microscopy in awake mice? Benefits: • Long-term longitudinal imaging • No interfering anesthesia • No need for control group Limitation: • Depth: only superficial layers of neocortex
  • 34. • Other cortical areas: motor, perirhinal, orbital cortex • Thalamocortical projections • Corticostriatal projections Primary somatosensory cortex (S1) connectivity
  • 35. Primary somatosensory cortex activation by drugs of abuse • Metabolic mapping • Activation marker c-Fos expression • fMRI
  • 36. • Changes in spine dynamics correlate with the synaptic activity at the spines and therefore serve as indicators of changes in neural circuitry produced by drugs of abuse • Persistence of the changes in spines point to a mechanism behind the long-term drug effects, including susceptibility to relapse • Changes in dendritic spine turnover are correlated with behavioral alterations Our hypotheses Thy1-YFP mice
  • 37. Effects of subchronic morphine - Spine turnover imaging protocol
  • 38. Week 1 M 1d M M M Week 3 M 1d M Week 2 MorphinePBS Effects of subchronic morphine - Spine turnover imaging protocol
  • 39. Week 1 M 1d M M M Week 3 M 1d M Week 2 MorphinePBS Effects of subchronic morphine - Spine turnover imaging protocol
  • 40. Week 1 M 1d M M M Week 3 M 1d M Week 2 MorphinePBS Effects of subchronic morphine - Spine turnover imaging protocol
  • 41. Week 1 M 1d M M M Week 3 M 1d M Week 2 MorphinePBS Effects of subchronic morphine - Spine turnover imaging protocol
  • 42. Week 1 M 1d M M M Week 3 M 1d M Week 2 MorphinePBS Effects of subchronic morphine - Spine turnover imaging protocol
  • 43. Week 1 M 1d M M M Week 3 M 1d M Week 2 MorphinePBS Effects of subchronic morphine - Spine turnover imaging protocol
  • 44. 100 mm Baseline, before PBS and Morphine 2 weeks after Morphine injections 100 mm Imaging stability (raw data comparison)
  • 45. • In vivo two-photon microscopy in non-anesthetized Thy1- YFP mice yields unbiased information of drug-induced changes in spine dynamics in the somatosensory cortex • Monitoring these changes over extended periods of time gives insight into drug-induced structural plasticity in the mammalian neocortex • Our pilot project with morphine demonstrates the feasibility of the procedures using the mobile home cage in the context of two-photon microscopy Summary of main points
  • 46. Electrophysiological recordings of cortical activity in head- restrained rodents in vivo Roustem Khazipov Directeur de Recherche, INSERM U901 Marat Minlebaev Charge de Recherche, INSERM U901
  • 47. 1. Overview of the extracellular and patch-clamp recording techniques (Khazipov) 2. Application of the Mobile HomeCage (Minlebaev) 3. Tips for stable electrophysiological recordings from the head-fixed animals in Mobile HomeCage What we are going to cover today:
  • 49. Intracortical recordings of the local field potentials Multi-electrode arrays Minlebaev & Khazipov, 2009
  • 50. Intracortical recordings of the local field potentials Minlebaev & Khazipov, 2009 • Local field potentials and multiple unit activity (MUA - spikes of individual neurons) can be recorded from different depth • Current source density (CSD) profile shows sinks and sources of the population events • MUA and cross-correlation analysis shows how neurons in different layers are activated during population events
  • 51. Multiple and single neuron action potentials Mitrukhina et al., 2014 • Extracellular recordings of neurons spikes with tetrodes enable to isolate spikes of individual neurons through cluster analysis • One tetrode may give a description of activity of several neurons
  • 52. Whole-cell recordings from a single neuron Minlebaev et al., 2011 • Whole-cell recordings from individual neurons enable to access synaptic correlates of the network activity • GABA and glutamate synaptic currents can be studied in isolation • In current clamp mode, firing of recorded cell and subthreshold conductances
  • 53. Cell-attached recordings • Cell-attached recordings enable to record spikes from individual neurons and • Currents through single ion channels • Reversal potential of GABA currents can be deduced from single GABA channel activity without altering intracellular chloride Tyzio et al., 2006
  • 54. 1. Overview of the extracellular and patch-clamp recording techniques (Khazipov) 2. Application of the Mobile HomeCage (Minlebaev) 3. Tips for stable electrophysiological recordings from the head-fixed animals in Mobile HomeCage What we are going to cover today:
  • 55. Spontaneous hippocampal activity in CA1 region of awake mice
  • 57. Cross-Frequency Coupling between Theta and Gamma frequency bands in Stratum Radiatum PAC measure = cfc Dataset = Structures = - Ampfreq/Hz- Phase freq / Hz - 5 10 15 20 25 30 35 40 0 10 20 30 40 50 60 70 80 90 0 0.005 0.01 0.015 0.02 0.025 0.03 0.035 0.04 0.045 0.05
  • 58. Multi-unit activity (StrR and StrPyr) phase lock to StrRad Local Field Potential 1s
  • 59. Current clamp whole cell in vivo recordings in the somatosensory cortex 2962029600295802956029540 Time (ms) Signal00 (mV) -90 -80 -70 -60 -50 -40 -30 -20 -10 0 3000029800296002940029200 Time (ms) Signal00 (mV) -90 -80 -70 -60 -50 -40 -30 -20 -10 0
  • 60. • Coordinates (atlases, papers) • Cortical surface better to keep wet (agarose better) • Ensure mechanical stability (make the hole in the skull as small as possible. Put dental cement close around the hole) • Minimize pulsations (put agar, silicone oil if needed) • Avoid multiple insertions (not more than 5-10 attempts through the same hole) • Vertical penetration (‘+’ extra cortical immobilization, ‘-‘ tissue damage; ~10 penetrations). • Pipette length depends on the recorded structure (shorter – better, otherwise extra capacitance; for striatum, up to 3 mm in mice) Patch-clamping in awake mice
  • 61. Thank You! For additional information on the Mobile HomeCage and methods for using this device in awake rodents with optical imaging or electrophysiological techniques please visit: http://www.neurotar.com/
  • 62. InsideScientific is an online educational environment designed for life science researchers. Our goal is to aid in the sharing and distribution of scientific information regarding innovative technologies, protocols, research tools and laboratory services.