Circadian expression of clock genes and clock-controlled genes in the rat retina

The circadian expression patterns of genes encoding for proteins that make up the core of the circadian clock were measured in rat retina using real-time quantitative PCR (qPCR). Transcript levels of several genes previously used for normalization of qPCR assays were determined and the effect of isc...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2005-04, Vol.330 (1), p.18-26
Main Authors: Kamphuis, Willem, Cailotto, Cathy, Dijk, Frederike, Bergen, Arthur, Buijs, Ruud M.
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Blotting, Northern
Circadian Rhythm
Clock genes
CLOCK Proteins
DNA Primers
Electrophoresis, Agar Gel
Gene expression
Ischemia
Male
Polymerase Chain Reaction
Quantitative PCR
Rat
Rats
Rats, Wistar
Reperfusion Injury - genetics
Retina
Retina - metabolism
RNA, Messenger - genetics
Trans-Activators - genetics
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Summary:The circadian expression patterns of genes encoding for proteins that make up the core of the circadian clock were measured in rat retina using real-time quantitative PCR (qPCR). Transcript levels of several genes previously used for normalization of qPCR assays were determined and the effect of ischemia–reperfusion on the expression of clock genes was studied. Statistically significant circadian changes in transcript levels were found for: Per2, Per3, Cry2, Bmal1, Rora, Rorb, and Rorc with changes ranging between 1.6- and 2.6-fold. No changes were found for Per1, Cry1, Clock, Rev-erb α, and Rev-erb β. Significant differences in transcript levels were observed for several candidate reference genes: HPRT, GAPDH, rhodopsin, and Thy1 and, consequently, the use of these genes for normalization purposes in qPCR or Northern blots may lead to erroneous conclusions. Ischemia–reperfusion leads to a persistent decrease of Per1 and Cry2, which may be related to the selective degeneration of amacrine and ganglion cells. We conclude that while all clock genes are expressed in the retina, only a few show a clear circadian pattern.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2005.02.118