Observation of the highly organized development of granule cells in rat cerebellar organotypic cultures

doi:10.1016/0006-8993(94)90161-9

Brain Research

Volume 641, Issue 2, 4 April 1994, Pages 319-327

https://doi.org/10.1016/0006-8993(94)90161-9 Get rights and content

Abstract

Cerebellar slices of 9-day-old rats were cultured for a week at the interface between air and a culture medium containing horse serum and hormone cocktail, and the development of granule cells was characterized morphologically. The typical layered structure of the cerebellar cortex was well preserved during the cultivation. Many granule cells in the external granular layer (EGL) proliferated actively within the early culture period. They were migrating downward at 3 days in vitro (DIV) and almost completed the migration to the internal granular layer (IGL) after 6 DIV. In the middle and upper molecular layer (ML), parallel fibers were elongated horizontally, which is the orientation 90° rotated compared to that in vivo. They formed synapses with Purkinje cell dendrites. Regional differences in synapse density and maturity existed which might reflect a gradient in progressive synapse formation comparable to that in vivo. Thus, a serial process of highly organized development of granule cells was realized for the first time in vitro, although some spatial or temporal modifications existed. Such a culture system could be a useful experimental model for the study of cellular and molecular mechanisms of spatiotemporally organized neuronal development.

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However, parallel fibers of the granule cells are oriented perpendicular to Purkinje dendrites and are better retained in the transversal plane (Heck & Sultan, 2002). However, granule cells are immature during the postnatal period and exhibit robust plasticity for regrowing axons and synapses with Purkinje dendrites in culture (Tanaka, Tomita, Yoshida, Yano, & Shimizu, 1994). We utilized the same equipment and technique described in the previous section with some modifications noted below.
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Observation of the highly organized development of granule cells in rat cerebellar organotypic cultures

Abstract

Exp. Cell Res.

Dev. Brain Res.

J. Neurosci. Meth.

Brain Res.

Dev. Biol.

Neuroscience

Neuroscience

Brain Res.

Brain Res.

Brain Res.

J. Neurosci. Meth.

Autoradiographic and histological studies of postnatal neurogenesis. III. Dating the time of production and onset of differentiation of cerebellar microneurons in rats

J. Comp. Neurol.

Postnatal development of the cerebellar cortex in the rat. I. The external germinal layer and the transitional molecular layer

J. Comp. Neurol.

Postntal development of the cerebellar cortex in the rat. II. Phases of the maturation of Purkinje cells and of the molecular layer

J. Comp. Neurol.

Mature Purkinje cells in cerebellar tissue culture: an ultrastructural study

J. Comp. Neurol.