title: The mouse cerebellar cortex in organotypic slice cultures: an in vitro model to analyze the consequences of mutations and pathologies on neuronal survival, development, and function.
abstract: Thin acute slices and dissociated cell cultures taken from different parts of the brain have been widely used to examine the function of the nervous system, neuron-specific interactions, and neuronal development (specifically, neurobiology, neuropharmacology, and neurotoxicology studies). Here, we focus on an alternative in vitro model: brain-slice cultures in roller tubes, initially introduced by Beat Gähwiler for studies with rats, that we have recently adapted for studies of mouse cerebellum. Cultured cerebellar slices afford many of the advantages of dissociated cultures of neurons and thin acute slices. Organotypic slice cultures were established from newborn or 10-15-day-old mice. After 3-4 weeks in culture, the slices flattened to form a cell monolayer. The main types of cerebellar neurons could be identified with immunostaining techniques, while their electrophysiological properties could be easily characterized with the patch-clamp recording technique. When slices were taken from newborn mice and cultured for 3 weeks, aspects of the cerebellar development were displayed. A functional neuronal network was established despite the absence of mossy and climbing fibers, which are the two excitatory afferent projections to the cerebellum. When slices were made from 10-15-day-old mice, which are at a developmental stage when cerebellum organization is almost established, the structure and neuronal pathways were intact after 3-4 weeks in culture. These unique characteristics make organotypic slice cultures of mouse cerebellar cortex a valuable model for analyzing the consequences of gene mutations that profoundly alter neuronal function and compromise postnatal survival.
authors.affiliation: Institut des Neurosciences Cellulaires et Intégratives, UMR 7168 LC2 CNRS/ULP, Department of Neurotransmission et Sécrétion Neuroendocrine, Centre de Neurochimie, 5 rue Blaise Pascal, F-67084 Strasbourg Cedex, France.
authors.forename: Etienne
authors.initials: E
authors.lastname: Lonchamp
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Dupont
authors.forename: Huguette
authors.initials: H
authors.lastname: Beekenkamp
authors.forename: Bernard
authors.initials: B
authors.lastname: Poulain
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Bossu
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D002525:Cerebellar Cortex / Q000166:cytology / Q000254:growth & development* / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D051379:Mice; D008959:Models, Neurological; D009154:Mutation / Q000235:genetics; D009434:Neural Pathways / Q000166:cytology / Q000254:growth & development* / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D009924:Organ Culture Techniques / Q000379:methods; D018408:Patch-Clamp Techniques