Cullingford T E, Bhakoo K, Peuchen S, Dolphin C T, Patel R, Clark J B
Department of Neurochemistry, Institute of Neurology, University College London, England, UK.
J Neurochem. 1998 Apr;70(4):1366-75. doi: 10.1046/j.1471-4159.1998.70041366.x.
We report the isolation, by RT-PCR, of partial cDNAs encoding the rat peroxisome proliferator-activated receptor (PPAR) isoforms PPAR alpha, PPAR beta, and PPAR gamma and the rat retinoid X receptor (RXR) isoforms RXR alpha, RXR beta, and RXR gamma. These cDNAs were used to generate antisense RNA probes to permit analysis, by the highly sensitive and discriminatory RNase protection assay, of the corresponding mRNAs in rat brain regions during development. PPAR alpha, PPAR beta, RXR alpha, and RXR beta mRNAs are ubiquitously present in different brain regions during development, PPAR gamma mRNA is essentially undetectable, and RXR gamma mRNA is principally localised to cortex. We demonstrate, for the first time, the presence of PPAR and RXR mRNAs in primary cultures of neonatal meningeal fibroblasts, cerebellar granule neurons (CGNs), and cortical and cerebellar astrocytes and in primary cultures of adult cortical astrocytes. PPAR alpha, PPAR beta, RXR alpha, and RXR beta mRNAs are present in all cell types, albeit that PPAR alpha and RXR alpha mRNAs are at levels near the limit of detection in CGNs. PPAR gamma mRNA is expressed at low levels in most cell types but is present at levels similar to those of PPAR alpha mRNA in adult astrocytes. RXR gamma mRNA is present either at low levels, or below the level of detection of the assay, for all cell types studied.
我们报告了通过逆转录聚合酶链反应(RT-PCR)分离出编码大鼠过氧化物酶体增殖物激活受体(PPAR)亚型PPARα、PPARβ和PPARγ以及大鼠视黄酸X受体(RXR)亚型RXRα、RXRβ和RXRγ的部分cDNA。这些cDNA被用于生成反义RNA探针,以便通过高灵敏度和高分辨力的核糖核酸酶保护分析,来检测发育过程中大鼠脑区相应的mRNA。在发育过程中,PPARα、PPARβ、RXRα和RXRβ的mRNA普遍存在于不同脑区,PPARγ的mRNA基本检测不到,而RXRγ的mRNA主要定位于皮层。我们首次证明,在新生脑膜成纤维细胞、小脑颗粒神经元(CGN)、皮层和小脑星形胶质细胞的原代培养物以及成年皮层星形胶质细胞的原代培养物中存在PPAR和RXR的mRNA。PPARα、PPARβ、RXRα和RXRβ的mRNA存在于所有细胞类型中,尽管PPARα和RXRα的mRNA在CGN中的水平接近检测极限。PPARγ的mRNA在大多数细胞类型中低水平表达,但在成年星形胶质细胞中的表达水平与PPARα的mRNA相似。在所研究的所有细胞类型中,RXRγ的mRNA要么水平较低,要么低于检测水平。