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

被引:79
作者
Kamphuis, W [1 ]
Cailotto, C [1 ]
Dijk, F [1 ]
Bergen, A [1 ]
Buijs, RM [1 ]
机构
[1] NORI KNAW, Glaucoma Res Grp, Dept Ophthalmogenet, Grad Sch Neurosci, NL-1105 BA Amsterdam, Netherlands
关键词
retina; gene expression; circadian rhythm; quantitative PCR; rat; clock genes; ischemia;
D O I
10.1016/j.bbrc.2005.02.118
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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, Bmall, Rora, Rorb, and Rorc with changes ranging between 1.6- and 2.6-fold. No changes were found for Perl, Cryl, Clock, Rev-erb alpha, and Rev-erb beta. Significant differences in transcript levels were observed for several candidate reference genes: HPRT, GAPDH, rhodopsin, and Thyl 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 Perl 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. (c) 2005 Elsevier Inc. All rights reserved.
引用
收藏
页码:18 / 26
页数:9
相关论文
共 45 条
[1]   Analysis of gene expression in the rat hippocampus using real time PCR reveals high inter-individual variation in mRNA expression levels [J].
Alfonso, J ;
Pollevick, GD ;
Castensson, A ;
Jazin, E ;
Frasch, ACC .
JOURNAL OF NEUROSCIENCE RESEARCH, 2002, 67 (02) :225-234
[2]   Hypothalamic integration of central and peripheral clocks [J].
Buijs, RM ;
Kalsbeek, A .
NATURE REVIEWS NEUROSCIENCE, 2001, 2 (07) :521-526
[3]   Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays [J].
Bustin, SA .
JOURNAL OF MOLECULAR ENDOCRINOLOGY, 2000, 25 (02) :169-193
[4]   Temporal-spatial characterization of chicken clock genes: circadian expression in retina, pineal gland, and peripheral tissues [J].
Chong, NW ;
Chaurasia, SS ;
Haque, R ;
Klein, DC ;
Iuvone, PM .
JOURNAL OF NEUROCHEMISTRY, 2003, 85 (04) :851-860
[5]   Circadian clock and microarrays: mammalian genome gets rhythm [J].
Delaunay, F ;
Laudet, V .
TRENDS IN GENETICS, 2002, 18 (12) :595-597
[6]   Validation of housekeeping genes for normalizing RNA expression in real-time PCR [J].
Dheda, K ;
Huggett, JF ;
Bustin, SA ;
Johnson, MA ;
Rook, G ;
Zumla, A .
BIOTECHNIQUES, 2004, 37 (01) :112-+
[7]   Differential effects of ischemia/reperfusion on amacrine cell subtype-specific transcript levels in the rat retina [J].
Dijk, F ;
van Leeuwen, S ;
Kamphuis, W .
BRAIN RESEARCH, 2004, 1026 (02) :194-204
[8]   Ischemia-induced changes of AMPA-type glutamate receptor subunit expression pattern in the rat retina: A real-time quantitative PCR study [J].
Dijk, F ;
Kraal-Muller, E ;
Kamphuis, W .
INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE, 2004, 45 (01) :330-341
[9]   Ischemia-induced alterations of AMPA-type glutamate receptor subunit. Expression patterns in the rat retina - an immunocytochemical study [J].
Dijk, F ;
Kamphuis, W .
BRAIN RESEARCH, 2004, 997 (02) :207-221
[10]   Arylalkylamine N-acetyltransferase gene expression in retina and pineal gland of rats under various photoperiods [J].
Engel, L ;
Vollrath, L ;
Spessert, R .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2004, 318 (04) :983-986