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[Neuroscience] Re: Cerebellar slices, advice?

Bill via neur-sci%40net.bio.net (by connelly.bill from gmail.com)
Sun Mar 25 20:41:54 EST 2007


Thanks for everyone's great comments.

Christian, if you'll excuse my inexcusable level of ignorance (I was
brought up on hippocampal slices, and the cerebellum still seems very
complicated), if I'm wanting to look at inhibition in cerebellar
granule cells; there inhibition comes solely (or nearly solely) from
golgi cells? So electrically evoked IPSCs would be best generated with
a fine electrode in the granule layer close to my patch electrode?
Also, if I'm after GABAergic inhibition, there is a significant
population of GlyRs out there, so strychnine is the order of the day?

Apologies for my naiveté. and thanks for your time.

On Mar 21, 10:53 pm, usene... from out-of-phase.de (Christian Wilms) wrote:
> We work with cerebellar slices on a regualr basis. The plane you decide
> to cut in depends on what you are looking for. Sagital slices will have
> intact climbing fibers and good Purkinje cell dendritic trees, but the
> parallel fibers will be severed. Transversal slices will leave long
> parts of the parallel fibers intact, but Purkinje cell dendrites will
> often be severed. I've also found that climbing fibers tend to be
> severely damaged in transversal slices. It seems to be a good idea to
> use fairly thick slices if you are cutting in the transversal plane (400
> microns), whereas thin slices (200 microns) work fine for the sagital
> plane.
>
> People using sagital slices usually stick to a basic preparation
> technique. We normally only use the Vermis. Hence, after removing the
> hind section of the skull I use a scapel to severe the hemispheres from
> the vermis as parallel to the midline as possible. Next I cut the Vermis
> free from the visual cortex with a clean cut perpendicular to the
> midline. For slicing I glue the explant down on one of the two sagital
> surfaces.
>
> We tend to use animals between P18 and 24. At this age mice and rats
> have fully developed parallel fibers and Purkinje cells. A collegue
> working on granule cells tends to use P60 animals - that would fit to
> Nadia's comments.
>
> Let me know if you have any specific questions.
>
> h2h, Christian




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