Organotypic culture of physiologically functional adult mammalian retinas.
BACKGROUND
The adult mammalian retina is an important model in research on the central nervous system. Many experiments require the combined use of genetic manipulation, imaging, and electrophysiological recording, which make it desirable to use an in vitro preparation. Unfortunately, the tissue culture of the adult mammalian retina is difficult, mainly because of the high energy consumption of photoreceptors.


RESULTS
We describe an interphase culture system for adult mammalian retina that allows for the expression of genes delivered to retinal neurons by particle-mediated transfer. The retinas retain their morphology and function for up to six days- long enough for the expression of many genes of interest-so that effects upon responses to light and receptive fields could be measured by patch recording or multielectrode array recording. We show that a variety of genes encoding pre- and post-synaptic marker proteins are localized correctly in ganglion and amacrine cells.


CONCLUSIONS
In this system the effects on neuronal function of one or several introduced exogenous genes can be studied within intact neural circuitry of adult mammalian retina. This system is flexible enough to be compatible with genetic manipulation, imaging, cell transfection, pharmacological assay, and electrophysiological recordings.
Koizumi
Amane
A
Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.
Zeck
Günther
G
Ben
Yixin
Y
Masland
Richard H
RH
Jakobs
Tatjana C
TC
D000200:Action Potentials; D025042:Amacrine Cells / Q000378:metabolism / Q000502:physiology; D000818:Animals; D005136:Eye Proteins / Q000235:genetics / Q000502:physiology; D005456:Fluorescent Dyes / Q000032:analysis; D018014:Gene Transfer Techniques; D009924:Organ Culture Techniques / Q000379:methods*; D018408:Patch-Clamp Techniques; D010957:Plasmids / Q000235:genetics; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000032:analysis; D012160:Retina / Q000502:physiology*; D012165:Retinal Ganglion Cells / Q000378:metabolism / Q000502:physiology
D005136:Eye Proteins; D005456:Fluorescent Dyes; D011993:Recombinant Fusion Proteins