Differential expression of mRNA and protein encoding retinal and pineal S-antigen during the light/dark cycle.
S-Antigen is a soluble cell protein unique to the retina and pineal gland. In the former, it is a well-characterized molecule that participates in light-induced signal transduction in photoreceptor cells. In the latter, the functional role is presently not known. The expression of S-antigen and its mRNA was examined in the rat retina and pineal gland throughout the diurnal cycle and with light interruption of the dark cycle. A cDNA for rat S-antigen was isolated from a pineal gland library to examine the mRNAs. A 1.7-kb mRNA for S-antigen was observed in both the pineal gland and the retina. Retinal S-antigen mRNA was expressed throughout the diurnal cycle and increased with light interruption of the dark cycle. In contrast, pineal gland S-antigen mRNA levels were detectable only during the dark and were absent preceding and during light. The phenotypic expression of immunoreactive S-antigen, identified with two S-antigen monoclonal antibodies (MAbs), MAb A9C6 and MAb C10C10, was analyzed by sodium dodecyl sulfate (SDS)-polyacrylamide gel (PAGE) and isoelectric focusing (IEF) electrophoresis. Immunoblot analysis of gels after SDS-PAGE revealed a single 46-kDa protein in retina. In contrast, two bands of approximately 43 and 46 kDa were identified in the pineal gland. Immunoblots of the retinal extracts separated by IEF electrophoresis revealed five S-antigen isomers, which vary quantitatively throughout the diurnal cycle and when light interrupted the dark cycle. Immunoblots of the pineal gland samples separated by IEF electrophoresis indicated that the pineal gland possesses four pineal gland-specific forms of S-antigen in addition to the five forms present in the retina. The differences observed in the mRNA and protein analyses suggest tissue-specific structural components for S-antigen in the retina and pineal gland that are not regulated in the same manner.
Craft
C M
CM
Department of Psychiatry, University of Texas Southwestern Medical School, Dallas 75235.
Whitmore
D H
DH
Donoso
L A
LA
D000595:Amino Acid Sequence; D000818:Animals; D000941:Antigens / Q000235:genetics*; D019393:Arrestin; D001483:Base Sequence; D004247:DNA / Q000302:isolation & purification / Q000378:metabolism*; D003624:Darkness; D004591:Electrophoresis, Polyacrylamide Gel; D005136:Eye Proteins / Q000235:genetics*; D015151:Immunoblotting; D007525:Isoelectric Focusing; D008027:Light; D008297:Male; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D010507:Periodicity; D010870:Pineal Gland / Q000276:immunology*; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012160:Retina / Q000276:immunology*; D012995:Solubility; D014020:Tissue Extracts / Q000032:analysis; D014867:Water
D000941:Antigens; D019393:Arrestin; D005136:Eye Proteins; D008565:Membrane Proteins; D012333:RNA, Messenger; D014020:Tissue Extracts; D014867:Water; D004247:DNA