Transcriptome comparison of murine wild-type and synaptophysin-deficient retina reveals complete identity.
Loss of synaptophysin, one of the major synaptic vesicle membrane proteins, is surprisingly well tolerated in knockout mice. To test whether compensatory gene transcription accounts for the apparent lack of functional deficiencies, comparative transcriptome analyses were carried out. The retina was selected as the most suitable tissue since morphological alterations were observed in mutant photoreceptors, most notably a reduction of synaptic vesicles and concomitant increase in clathrin-coated vesicles. Labeled cRNA was prepared in triplicate from retinae of age- and sex-matched wild-type and mutant litter mates and hybridized to high-density microarray chips. Only three differentially expressed RNAs were identified in this way, one of which was synaptophysin. Further validation by quantitative RT-PCR could only corroborate the results for the latter. We therefore conclude that, despite the distinct morphological phenotype, no significant changes in gene expression are detectable in synaptophysin-deficient animals and that therefore compensatory mechanisms are either pre-existent and/or act at the posttranscriptional level.
Bai
Lin
L
Department of Anatomy and Cell Biology, Johannes Gutenberg University, Becherweg 13, 55128 Mainz, Germany.
Spiwoks-Becker
Isabella
I
Leube
Rudolf E
RE
D000818:Animals; D002966:Clathrin / Q000378:metabolism; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation; D043326:Glucan 1,3-beta-Glucosidase; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D046228:Microarray Analysis / Q000379:methods; D046529:Microscopy, Electron, Transmission / Q000379:methods; D010786:Photoreceptor Cells / Q000378:metabolism / Q000648:ultrastructure; D012333:RNA, Messenger / Q000096:biosynthesis; D012160:Retina / Q000378:metabolism* / Q000648:ultrastructure; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013572:Synaptic Vesicles / Q000378:metabolism / Q000648:ultrastructure; D016708:Synaptophysin / Q000172:deficiency*
D002966:Clathrin; D005136:Eye Proteins; D012333:RNA, Messenger; D016708:Synaptophysin; D043326:Glucan 1,3-beta-Glucosidase