Spontaneous Synaptic Activity in an Organotypic Culture of the Mouse Retina

Abstract

The different decay kinetics of GABA(A)Rs expressed in amacrine cells of geph(+/+) and of geph(-/-) retinas suggests that these cells express at least two types of GABA(A)R subtypes. In amacrine cells of geph(-/-) mice, a specific GABA(A)R subtype that may contain the alpha2 subunit, is impaired by the absence of gephyrin, whereas other GABA(A)Rs appear to function normally.

Nicotinamide Adenine Dinucleotide-Dependent Binding of the Neuronal Ca2+Sensor Protein GCAP2 to Photoreceptor Synaptic Ribbons

“…SLF-mediated GCAP2-EGFP ( A–C ) heterologous expression labels the entire photoreceptor from the inner segments to the synaptic terminals (arrows) in the OPL. As generally observed by us and other groups (Fischer et al, 2000; Perez-Leon et al, 2003; Zhang et al, 2008), outer segments are absent from explant preparations. In analogy to GCAP2-EGFP expression, infection with EGFP-SLF virus also leads to labeling of the entire photoreceptor ( D ) in retina explant culture.…”

“…Preparation of organotypic cultures was performed mostly as described previously (Fischer et al, 2000; Pérez-León et al, 2003; Zhang et al, 2008), with some modifications. Briefly, freshly isolated eyes enucleated from adult mice housed under ambient light conditions were immediately immersed into ice-cold RPMI 1640 (supplemented with 10% fetal calf serum, 10 mM HEPES, 2 mM L-glutamine, 1 mM sodium pyruvate, 50 μ M β ME, 100 U/ml penicillin, and 100 μ g/ml streptomycin).…”

A novel organotypic culture model of the postnatal mouse retina allows the study of glutamate-mediated excitotoxicity

“…From the early 1980s there had been some reports of organotypic cultures of retina tissue on collagen substrate (Smalheiser et al, 1981;Carri and Ebendal, 1983). Perez-Leon et al (2003) presented that they cultured mouse retina onto sterile nitrocellulose filters which were then attached to coverslips and inserted into screw-top culture tubes and Schultz-Key et al (2002) sectioned rat retina into slices first with a McIIwain tissue chopper and then cultured them onto membranes or glass coverslips. We cultured retinal tissue directly on poly-d-lysine/laminin coated coverslips because of poly-d-lysine's property to enhance cell attachment to plastic and glass surfaces for many cell types and laminin's established property as a substrate for growth and differentiation of many cell types.…”

Roller Culture of Free-Floating Retinal Slices: A New System of Organotypic Cultures of Adult Rat Retina

“…However, many studies showed complete or partial degeneration of RGCs as a result of axotomy in organotypic retina cultures from newborn and postnatal animals. In neonatal organotypic retinal cultures, partial degeneration of RGCs was observed after 24 h in vitro [6,7,35] , while some RGCs were still present after 4 weeks in culture [10] . In roller drum retinal slice cultures of 5-to 7-day-old rats embedded in a plasma clot, most of the RGCs degenerated, and only few neurons expressing Thy-1 were found [13] .…”

“…Stationary fl atmount cultures of embryonic and early postnatal retinas, sometimes embedded in a plasma clot or in a three-dimensional collagen gel, are used mostly for the study of retinal ganglion cell (RGC) axonal growth as well as of factors which modify axonal regeneration [3][4][5] . Organotypic cultures of neonatal or postnatal mouse and rat retinas with preservation of the organotypic cell structure were grown on semipermeable membranes and Millipore fi lter membranes [6][7][8][9][10] . Survival of retinas of postnatal and adult mice was obtained in organ cultures grown in serum-free medium [9,11] .…”