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(Investigative Ophthalmology and Visual Science. 2003;44:1376-1387.)
© 2003 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.02-0702
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Spontaneous Synaptic Activity in an Organotypic Culture of the Mouse Retina

Jorge Pérez-León,1 Moritz J. Frech,1 Jörn E. Schröder,1 Frauke Fischer,1 Matthias Kneussel,2 Heinz Wässle,1 and Kurt H. Backus1

1From the Departments of Neuroanatomy and 2Neurochemistry, Max-Planck-Institute for Brain Research, Frankfurt/M., Germany.

PURPOSE. Many strains of mutant mice die at birth, when the retina is still very immature. The retinas of such mice can be studied in organotypic cultures. After a preceding anatomic study of the synaptic development, the electrical activity of the synaptic circuits within such cultures was studied in wild-type and gephyrin-deficient mice.

METHODS. Organotypic cultures of newborn mouse retinas were grown for 14 days in vitro. Spontaneous postsynaptic currents (sPSCs) of amacrine cells were measured by using the whole-cell configuration of the patch-clamp technique. GABAergic and glycinergic currents that were isolated with specific antagonists, and retinas from wild-type (geph+/+) and gephyrin-deficient (geph-/-) mice were compared.

RESULTS. Rapidly decaying sPSCs that were blocked by kynurenic acid were mediated by ionotropic glutamate receptors, whereas sPSCs with significantly higher peak amplitudes and slow-decay kinetics were identified as spontaneous inhibitory postsynaptic currents (sIPSCs) mediated by {gamma}-aminobutyric acid type A receptors (GABAARs) and glycine receptors (GlyRs). In gephyrin-deficient (geph-/-) cultures, we found no sIPSCs mediated by GlyRs. sIPSCs mediated by GABAARs expressed in amacrine cells of geph-/- retinas decayed significantly faster than GABAergic sIPSCs recorded in amacrine cells of geph+/+ retinas.

CONCLUSIONS. The different decay kinetics of GABAARs expressed in amacrine cells of geph+/+ and of geph-/- retinas suggests that these cells express at least two types of GABAAR subtypes. In amacrine cells of geph-/- mice, a specific GABAAR subtype that may contain the {alpha}2 subunit, is impaired by the absence of gephyrin, whereas other GABAARs appear to function normally.






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