Organotypic cerebellar cultures: apoptotic challenges and detection.

Content Provider	Europe PMC
Author	Hurtado de Mendoza, Tatiana Balana, Bartosz Slesinger, Paul A. Verma, Inder M.
Copyright Year	2011
Abstract	Organotypic cultures of neuronal tissue were first introduced by Hogue in 1947 1,2 and have constituted a major breakthrough in the field of neuroscience. Since then, the technique was developed further and currently there are many different ways to prepare organotypic cultures. The method presented here was adapted from the one described by Stoppini et al. for the preparation of the slices and from Gogolla et al. for the staining procedure 3,4.A unique feature of this technique is that it allows you to study different parts of the brain such as hippocampus or cerebellum in their original structure, providing a big advantage over dissociated cultures in which all the cellular organization and neuronal networks are disrupted. In the case of the cerebellum it is even more advantageous because it allows the study of Purkinje cells, extremely difficult to obtain as dissociated primary culture. This method can be used to study certain developmental features of the cerebellum in vitro, as well as for electrophysiological and pharmacological experiments in both wild type and mutant mice.The method described here was designed to study the effect of apoptotic stimuli such as Fas ligand in the developing cerebellum, using TUNEL staining to measure apoptotic cell death. If TUNEL staining is combined with cell type specific markers, such as Calbindin for Purkinje cells, it is possible to evaluate cell death in a cell population specific manner. The Calbindin staining also serves the purpose of evaluating the quality of the cerebellar cultures.
Journal	Journal of Visualized Experiments : JoVE
PubMed Central reference number	PMC3197101
Issue Number	51
PubMed reference number	21633327
e-ISSN	1940087X
DOI	10.3791/2564
Language	English
Publisher	MyJove Corporation
Publisher Date	2011-05-17
Access Restriction	Open
Rights License	This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/ Copyright © 2011, Journal of Visualized Experiments
Subject Keyword	Neuroscience Issue 51 Cerebellum Organotypic Fas Apoptosis Purkinje cell
Content Type	Text
Resource Type	Article
Subject	Immunology and Microbiology Chemical Engineering Neuroscience Biochemistry, Genetics and Molecular Biology