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Organotypic slice culture of the mammalian retina

Published online by Cambridge University Press:  02 June 2009

Andreas Feigenspan
Affiliation:
Max-Planck Institut für Hirnforschung, Neuroanatomische Abteilung, Deutschordenstrasse 46, W 6000 Frankfurt, Germany.
Joachim Bormann
Affiliation:
Max-Planck Institut für Hirnforschung, Neuroanatomische Abteilung, Deutschordenstrasse 46, W 6000 Frankfurt, Germany.
Heinz Wässle
Affiliation:
Max-Planck Institut für Hirnforschung, Neuroanatomische Abteilung, Deutschordenstrasse 46, W 6000 Frankfurt, Germany.

Abstract

Vertical slices of 6-day postnatal (P6) rat retina were cut at a thickness of 100 μm and cultured using the roller-tube technique. After 14–21 days in vitro there was significant distortion of normal retinal architecture, but localized areas of the slices showed the typical pattern of layering of mature retina. The following immunocytochemical markers were used to characterize the different retinal cell types: antibodies against protein kinase C (PKC), calcium binding protein (CabP 28kD), neurofilaments (NF), glia-specific antibodies (GFAP, vimentin), and transmitter-specific antibodies (GABA, TH). The expression of these markers was compared in P6 retina, adult retina, and slice culture. To further characterize the cultured cells, patch-clamp recordings were performed in combination with intracellular injection of Lucifer Yellow (LY). Transmitter-and voltage-gated membrane currents were recorded from morphologically identified neurons. The experiments show that a mammalian slice culture can be used to study differentiation and function of retinal cell types.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1993

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