Tweak Your Technique

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Electrophysiology 101. August 19, 2011. Kevin Lee. What can be studied with electrophysiology?. Single channel behaviour. in expression systems: HEK293, CHO’s, X. . Laevis. . oocytes. Intrinsic properties of excitable cells. ID: 567099 Download Presentation

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Tweak Your Technique




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Presentations text content in Tweak Your Technique

Slide1

Tweak Your TechniqueElectrophysiology 101

August 19, 2011

Kevin Lee

Slide2

What can be studied with electrophysiology?

Single channel behaviour

in expression systems: HEK293, CHO’s, X.

Laevis

oocytes

Intrinsic properties of excitable cells

neurons,

myocytes

Synaptic transmission

neuromuscular junction, neuronal synapses

Slide3

Slide4

The hippocampus

Slide5

Field recordings

Slide6

LTP/LTD

Slide7

Whole-cell Recordings

Slide8

Patch-Recording Methods

Slide9

Whole-cell recordings

Current-clamp vs Voltage-clamp

“EPSP”

“EPSC”

Slide10

Whole-cell recordings

Current-clamp vs Voltage-clamp

Slide11

Voltage-clamp

Slide12

mEPSCs

(minis)

mEPSC

amplitude reflects synaptic AMPAR content

Beique

et al. (PNAS, 2006)

Slide13

Beique

et al. (PNAS, 2006)

mEPSCs

(minis)

mEPSC

frequency can reflect:

- total number of synapses

-

presynaptic

function

Slide14

Release probability (Pr) of glutamate

Changes in frequency of mEPSCs is generally believed to reflect changes in Pr.“Paired-Pulse” electrical stimulation can also provide information about Pr.

Beique

et al. (PNAS, 2006)

Slide15

What about Molecular interventions?

Slide16

Biolistic transfection of cortical neurons in organotypic slices

Beique

& Andrade (J.

Physiol

, 2002; cover photo)

Slide17

Biolistic transfection of cortical neurons in organotypic slices

Slide18

Biolistic transfection of cortical neurons in organotypic slices

Barria

& Malinow (Neuron, 2002)

Cull-Candy et al. (Science STKE, 2004)

Slide19

Drawbacks to electrical stimulation:

Difficult to isolate individual synaptic events

Spatial Parameters left unknown

Slide20

2P-Uncaging of MNI-Glutamate

Slide21

2P-Uncaging of MNI-Glutamate

Béïque et al., PNAS 2006

Uncaging

of MNI-

Glu activates both AMPA and NMDA receptors

High spatial resolution of glutamate

uncaging

Slide22

Single Synapse Ca

2+

Imaging

Time

1

2

1

2

Uncage

Simultaneous Glutamate

Uncaging

and Calcium Imaging

Slide23

Fundamental dendritic properties probed by glutamate uncaging and calcium imaging

Losonczy

& Magee (Neuron, 2006)

Slide24

Fundamental

dendritic properties probed by glutamate uncaging and calcium imaging

Losonczy

& Magee (Neuron, 2006)

Slide25

Experimental Wizardry to Envy

Kravitz

et al. (Nature, 2009)

Slide26

Experimental Wizardry to Envy

Kravitz

et al. (Nature, 2009)

Slide27

Multiple Simultaneous Recordings in Slices

Olah

et al. (Nature, 2009)

Slide28

The

End


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