Methods in Neuroscience Flashcards
what questions can we ask?
Santiago Ramon y Cajal said we can ask how but not why questions.
But how questions can be asked on many different levels.
Cellular – how do neurons communicate?
Anatomical – how are different areas interconnected (cytoarchitectonics)?
Functional – how do certain areas mediate certain behaviors?
- How do we know occipital cortex plays big role in vision?
Systemic – how do disease processes affect brain function?
- What does Parkinson do, Alzheimer’s do?
List some methods
Cellular – histology and staining.
- Ways to prepare tissue so they can be seen under microscope
Anatomical – autopsy of healthy and diseased brains.
Functional methods; human (natural/rarely induced - stroke, bullet wound, car accident etc.) and animal (induced - specific areas) brain lesions.
neural stimulation – animal studies (single units) and human studies (disruption to networks – TMS).
- TMS = transcranial magnetic stimulation - way of temporary lesioning or existing cells
Systemic – neuroimaging – the brain in action.
Who is Santiago Ramon y Cajal?
father of modern day neuroscience
Lesion methods
removal of tissue (ablation), radio frequency lesions, chemical lesions(can target certain transmitters), reversible chemical lesions, cooling(reversible, can use animal as own control subject).
What is a lesion?
is removal of tissue but can be done in different ways
Lesion studies
Assume that if cognition X is disrupted by a lesion to a brain area Y then region Y “supports” function X.
If an area of the brain like occipital cortex if lesioning it affects vision then then that area is linked to vision
Modular concept of brain organization – a place for everything and everything in its place.
What shouldn’t be forgotten?
Shouldn’t forget that many functions are highly distributed throughout the brain!
Damaging a particular area may affcet behaviour but doesn’t mean only that area is involved in that behviour a network could be involved
Animal lesion studies
experimental ablation.(removal of tissue)
caution in interpretation – change in behavior after a lesion could be due to many things:
a) the loss of function supported by that particular region.
b) the loss of a sub-component necessary for the behavior caused by disruption to the network.
c) co-existing behavioural changes unrelated to the particular region.
- Stress of surgery or something else
Animal lesion models
Lesions may also change behaviour (adaptation to the deficit).
Monkeys deprived of sensory feedback to one limb will favour use of the other limb.
When deprived of sensory feedback to both limbs they will not favour one over the other.
- haven’t knocked out the ability for that monkey to use that limb, but it’s had a preference now for using the other limb
- but if you knock out sensory feedback, it uses both
Methodological limitation
correspondence between animals and humans may be low (e.g., right parietal lesions in monkeys does not produce hemi spatial neglect).
Animal lesion studies types
- high frequency radio pulse
- chemical lesions
- excitotoxic lesion
- reversible lesions
High frequency radio pulse
destroys everything.
Chemical lesions
targets specific neurotransmitters neurons.
Excitotoxic lesion
spares axons that are passing through area.
- think: head is lesion (sparing axons passing through)
Reversible lesions
anesthetic or cooling
- cooling a section of the brain
(neurons not firing as much - almost temporarily inactivating that area of the brain).
Reversible Lesions: Design Issues
Within-subjects designs
test with the lesion and without
increased statistical power
Subject is testing as its own control
Test same animal with brain cool and brain normal
Disadvantage of reversible lesions
don’t euthanize the animal, stain the brain, and determine that the lesion was in the desired location.
- hard to know if you hit the right area
Stereotaxic Surgery
- helmet like mount on a person’s shaved head
- purpose: so surgeon can use a map of the brain to know where they’re doing the implantation
- use a particular seam in the skull (a SUTURE) as a landmark.
- BREGMA - the junction of the coronal and sagittal sutures (fontanel in infants).
- BREGMA
the junction of the coronal and sagittal sutures (fontanel in infants, soft spot, young infant baby hasn’t fused yet)
Skull Sutures: Targets for Lesion or Electrode Placement
- Cut scalp of rat open so we are looking at the skill
- Use bregma as 0.00 point
- Lower an electrode so that the tip touches the edge of the skull
- We take a recording of that
- If we use that as our 0 point, we can then use a map/altus of the brain to figure out how far down/left or right, forward or back etc. we have to go to hit the area we want to hit
- Have to figure out where to drill the hole & how far down do we have to send it in order to target that specific area of interest
Stereotaxic Atlas
like goggle maps of brain
Breaks brain down into diff quadrant
So you can figure out how to adjust stereotaxic device to leison the areas your testing
Rats are anesthetized, place the rat so that ear bars go into ears and tightly lock the skull into position, bite bar is put Infront of front teeth so that skull is in position, make sure its in level, holes are drilled and the tip of the electrode is sunk down to the level specified in the stereotaxic atlas.
Current is turned on, ablating the structure.
Rat is sutured and is tested after a couple of days.
Lesion group
anesthesia
incision
lesion
suture and stitches
Sham group
anesthesia
incision
sutures and stitches
Note the incision in the sham group includes lowering the electrode or cannula to the appropriate depth.
control animals must undergo all aspects of the lesion study except the lesion itself.