期刊
JOURNAL OF NEUROTRAUMA
卷 32, 期 22, 页码 1777-1788出版社
MARY ANN LIEBERT, INC
DOI: 10.1089/neu.2014.3784
关键词
aging; control genes; housekeeping; normalization; quantitative real-time RT-PCR; reference genes; traumatic brain injury
资金
- German Research Foundation [DFG TH1430/3-1, CRC1080/A9]
- Federal Ministry of Education and Research [BMBF 01EO1003]
To prevent methodological errors of quantitative PCR (qPCR) normalization with reference genes is obligatory. Although known to influence gene expression, impact of age on housekeeping gene expression has not been determined after acute brain lesions such as traumatic brain injury (TBI). Therefore, expression of eight common control genes was investigated at 15min, 24h, and 72h after experimental TBI in 2- and 21-month-old C57Bl6 mice. Expression of (2)-microglobulin (B2M), -actin (ActB), and porphobilinogen deaminase (PBGD) increased after TBI in both ages. 2M demonstrated age-dependent differences and highest inter- and intragroup variations. Expression of cyclophilin A, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), hypoxanthine ribosyltransferase (HPRT), S100B, and 18SrRNA remained stable. Cyclophilin A and HPRT demonstrated strongest inter- and intragroup stability. The data indicate that the expression of most but not all control genes is stable during aging. The correct choice of housekeeping genes is of key importance to ensure adequate normalization of qPCR data. With respect to insult and age, normalization strategies should consider cyclophilin A as a single normalizer. Normalization with two reference genes is recommended with cyclophilin A and HPRT in young mice and in mixed age studies and with cyclophilin A and GAPDH in old mice. In addition, the present study suggests not to use (2)-microglobulin, -actin or PBGD as single control genes because of strong regulation after CCI in 2- and 21-month-old mice.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据