Differential expression of cone opsin mRNA levels following experimental retinal detachment and reattachment.
OBJECTIVE
To identify changes in S- and M-sensitive cone opsin gene expression following retinal detachment (RD) and reattachment.


METHODS
Cat retinas were detached for 1, 3, 7, or 28 days, or reattached after 1 h, 1 day, or 3 days of RD and fixed in 4% paraformaldehyde. Pieces of mid-peripheral retina were removed from the same region of each detached, normal (attached), and reattached retina and embedded in paraffin. Paraffin sections (8 mm) were processed for in situ hybridization using S- or M-cone opsins, rod opsin, or phosducin riboprobes in vitro transcribed from cat partial cDNAs. Labeled cells were counted to obtain the number of labeled cells/mm retina.


RESULTS
The number of cells labeled with the anti-sense cone opsin riboprobes, and the intensity of this label, decreased after RD. The number of cones labeled with the anti-sense S-opsin riboprobe decreased to 42% of normal at 3 days of RD. The number of M-opsin mRNA-positive cones decreased to 4% of normal at 3 days of RD. The number of cells positive for M-opsin or S-opsin mRNA recovered to near normal levels after reattachment. Phosducin and rod opsin mRNA labeling was near normal in surviving rod photoreceptors after RD.


CONCLUSIONS
Cones and rods behave differently after detachment. There are significant obstacles to overcome in order to study the responses of cones after RD because surviving cells no longer label with antibodies used as cone markers in normal retina. The results of this study show that: (1) After RD, surviving cones decrease their expression of opsin mRNA while rods do not; (2) Upon reattachment of the retina, the cones once again begin to express their opsins; (3) Most cones survive short-term detachments; and (4) Defects in cone-based vision after reattachment may not be based mainly on the loss of cones but due to other changes in these cells, for example, reduced phototransduction and/or changes in synaptic connectivity to second order neurons.
Rex
Tonia S
TS
Neuroscience Research Institute and Department of Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, Santa Barbara, CA. trex@mail.med.upenn.edu
Lewis
Geoffrey P
GP
Geller
Scott F
SF
Fisher
Steven K
SK
D000818:Animals; D002415:Cats; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000032:analysis; D004195:Disease Models, Animal; D015870:Gene Expression; D017403:In Situ Hybridization; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism*; D017949:Retinal Cone Photoreceptor Cells / Q000378:metabolism*; D012163:Retinal Detachment / Q000378:metabolism* / Q000601:surgery; D017948:Retinal Rod Photoreceptor Cells / Q000378:metabolism; D017299:Rod Opsins / Q000235:genetics*
D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger; D017299:Rod Opsins