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BMS 248 - Neural circuits > Modern methods in neuroscience II > Flashcards

Modern methods in neuroscience II Flashcards

1
Q

What did Cajal and Golgi develop?

A

Ways to label cells

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2
Q

What was the difference between labelling cells in Cajal and Golgi techniques?

A

Golgi:
- Spare labelling (not all of the cell)

Cajal:
- Fine details

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3
Q

With Golgi labelling, what can be seen?

A

The MORPHOLOGY of the NERUONS

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4
Q

With Cajal labelling, what can be seen?

A

Individual DENDRITES

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5
Q

What are the problems with Golgi and Cajal methods of looking at the morphology of neurons?

A
  • Can’t label INDIVIDUAL neurons

- Cannot combine ELECTOPHYSIOLOGY and MORPHOLOGY assesment in the same cell (neurons are dead)

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6
Q

What do we want to study in order to understand about new neuronal types?

A

1) Morphology of the neurons
2) Map connections
3) Activity of the neurons (what stimuli activate them)
4) Theoretical study

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7
Q

What is the electrophysiochemical method used to study neurons?

A

Patch clamp technique

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8
Q

What can be used to measure morphology and electrophysiology in one experiment?How?

What is the advantage of this?

A

Patch clamp technique with flurorescence in the electrode
- Fluorescence will diffuse into the cell

Advantage:

  • The neuron is still alive
  • Can see how the morphology/electrophysiology changes?
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9
Q

What are the problems with patch clamp with fluorescence to study neurons?

A

1) Cannot label may cells
2) Limited ability to label a specific cell type
3) Limited ability to label cellular compartments (eg. dendrites, specific receptors)
4) Still LIMITED ability for live labelling

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10
Q

Why is there limited ability to label cellular compartments (eg. receptors) in the cell with patch clamp/fluorescence?

A

Use a SYNTHETIC DYE - goes everywhere in the cell

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11
Q

Using the patch clamp/fluorescence technique, why is there still limited ability for live labelling?

A

In most cases, method is applied to SLICES of tissue

Complicated with live animals

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12
Q

How are the problems with patch clamp/fluorescence techniques overcome?

A

Using GFP

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13
Q

What is GFP stimulated by?

What does it emit?

A

Blue light

Emits green light

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14
Q

What are RFP, YFP and CFP?

A

Modifications of GFP that have different colours

different excitatory and emission wavelengths of light

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15
Q

What can GFP be used for?

A
  • To label cells (morphology)

- Follow MIGRATION of cells and the PROJECTIONS of the neurons

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16
Q

How can GFP be used to study the activity of neurons?

A

Use a modification of GFP (GCaMP)

17
Q

What is GCaMP?

What is the structure?

A

GFP based calcium indicator

GFP fused to 2 calcium binding proteins (M13 and Calmodulin)

18
Q

What happens in GCaMP in the presence of calcium?

What does this cause?

A

M13 and Calmodulin interact and STABILSE the GFP

Causes the GFP to become brighter

19
Q

What happens when the neuron is active when using GCaMP and why>

A

Calcium channels open
Calcium levels in the cell rise
Causes more M13 and calmodulin interaction
Causes GFP to be brighter

20
Q

What is the difference between wide field and confocal microscopy?

A

Wild field - collects light from above and below the focal plane

Confocal - Pinhole rejects light NOT coming FROM the FOCAL plane (reject light from above and below the FP)

21
Q

What does confocal microscopy result in?

A

A large increase in SPATIAL resolution

22
Q

What is the problem with using GCaMP to study function of neurons?

How are these problems overcome?

A
  • Animal is sedated using Na channel blockers
  • Neurons spike less

OR

  • Animal is stressed
  • Release of other molecules in the brain
  • Animal doesn’t perform the behaviour that is normally does

Overcome by using:

  • Virtual reality
  • Freely moving animal
23
Q

Describe the virtuality technique of studying the function of neurons

A
  • Mouse on a ball that moves in a certain direction when the mouse moved
  • Head is fixed
  • Turn of the ball leads to a turn of the field of view in that direction
24
Q

Describe the freely moving mice teqnique of studying the function of neurons

A
  • Tiny fluorescent microscope placed directly into the skull of the mouse
  • Image is delivered via light guide
25
Q

How are channel-rhodopsin and halorhodopsin used?

A

Introduced into a cell

Used to modify neuronal activity

26
Q

How is channel-rhodopsin activated?

A

Using blue light

27
Q

What happens when channel-rhodopsin is activated?

A

Depolarisation of the membrane

28
Q

What type of channel is channel-rhodopsin?

A

Non-specific cation channel

29
Q

What activates halorhodopsin?

A

Yellow light

30
Q

What ions pass through halorhodopsin?

A

Cl

31
Q

What happens what halorhodopsin is activated?

A

The cell is hyperpolarised

BMS 248 - Neural circuits (16 decks)

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