Retina expresses a novel variant of the ryanodine receptor.
Calcium released from intracellular stores via the ryanodine receptor (RyR) mediates a variety of signalling processes. We previously showed that retina expresses the three known types of RyR, but retinal membrane preparations exhibit unique characteristics such as Ca2+-independent [3H]ryanodine-binding and inhibition by caffeine. We have heretofore suggested that the major retinal RyR isoform is novel. The present study aimed to identify this receptor isoform and to localize RyR in mammalian retina. Immunoblotting with specific and pan-antibodies showed that the major retinal RyR has a mobility similar to that of RyR2 or RyR3. Real-time PCR revealed that the major type is RyR2, and RT-PCR followed by sequencing showed a transcript that encodes a protein with approximately 99% identity to RyR2, yet lacking two regions of seven and 12 amino acids and including an additional insertion of eight amino acids. An antibody against RyR2 localized this type to somas and primary dendrites of most retinal neurons. An antibody against RyR1 localized RyR to most somas but also revealed staining in photoreceptor outer segments, concentrated on the disk membranes at their rim. The ryanodine-binding properties and the electrophoretic mobility of RyR from the outer segments were similar to those of the whole retinal preparation. The results thus identify a novel variant of RyR2 which can contribute to regulating photoreceptor Ca2+ concentrations. The restricted localization of the outer segment RyR to the disk rim suggests that its activation mechanism involves a coupling between retinal RyR and the cGMP-gated channel.
Shoshan-Barmatz
Varda
V
Department of Life Sciences, Ben Gurion University of the Negev, Beer Sheva, 84105, Israel. vardasb@bgu.ac.il
Zakar
Miri
M
Shmuelivich
Fania
F
Nahon
Edna
E
Vardi
Noga
N
D000818:Animals; D002417:Cattle; D003001:Cloning, Molecular / Q000379:methods; D015870:Gene Expression / Q000502:physiology*; D016253:Microscopy, Immunoelectron / Q000379:methods; D008954:Models, Biological; D011817:Rabbits; D051381:Rats; D012160:Retina / Q000378:metabolism* / Q000648:ultrastructure; D012433:Ryanodine / Q000378:metabolism; D019837:Ryanodine Receptor Calcium Release Channel / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D016415:Sequence Alignment / Q000379:methods; D013347:Subcellular Fractions / Q000378:metabolism / Q000648:ultrastructure; D014316:Tritium / Q000378:metabolism
D019837:Ryanodine Receptor Calcium Release Channel; D014316:Tritium; D012433:Ryanodine