Erdmann B, Gerst H, Lippoldt A, Bülow H, Ganten D, Fuxe K, Bernhardt R
Max-Delbrück-Center for Molecular Medicine (MDC), Berlin-Buch, Germany.
Brain Res. 1996 Sep 9;733(1):73-82. doi: 10.1016/0006-8993(96)00540-9.
Cytochrome P45011B1 (11 beta-hydroxylase) was detected in the brain of male rats by in situ hybridization methods. Normal Sprague-Dawley rats were compared to the transgenic strain TGR(mRen2)27, characterized by the expression of the murine Ren-2d renin gene and the development of severe hypertension. Specific riboprobes were generated by in the vitro transcription of a 152 base-pair long cDNA template 35S-labeled riboprobes were hybridized to cryostat sections from adrenal glands and from two different levels of the brain using standard protocols and varying washing conditions. After exposure of the radiolabeled sections to X-ray film, the signals were quantified and compared. Following autoradiography and counterstaining, cytochrome P45011B1 mRNA was clearly localized in the zona fasciculata/reticularis of the adrenal cortex and in distinct layers of the cerebral cortex. High signal densities were obtained in the layers II-IV of the neocortex and in the layer II of the piriform cortex, although the concentrations of cytochrome P45011B1 mRNA were remarkably lower in the central nervous system as compared to adrenal glands. As revealed by the semi-quantitative analysis, there was a slight increase in adrenal 11 beta-hydroxylase mRNA in the transgenic rats, whereas the brain seems to express nearly the same amount of this enzyme in both strains. The cytochrome P45011B1 mRNA expression in distinct cells, probably nerve cells, and especially in regions with high densities of glucocorticoid receptors points to a possible function of brain derived corticosterone in receptor activation.
通过原位杂交方法在雄性大鼠脑中检测到细胞色素P45011B1（11β-羟化酶）。将正常的Sprague-Dawley大鼠与转基因品系TGR（mRen2）27进行比较，该品系的特征是表达小鼠Ren-2d肾素基因并发展为严重高血压。通过对152个碱基对长的cDNA模板进行体外转录产生特异性核糖探针，使用标准方案和不同的洗涤条件，将35S标记的核糖探针与来自肾上腺和脑的两个不同水平的冰冻切片杂交。将放射性标记的切片暴露于X射线胶片后，对信号进行定量和比较。经过放射自显影和复染后，细胞色素P45011B1 mRNA明显定位于肾上腺皮质的束状带/网状带以及大脑皮质的不同层。在新皮质的II-IV层和梨状皮质的II层获得了高信号密度，尽管与肾上腺相比，细胞色素P45011B1 mRNA在中枢神经系统中的浓度明显较低。半定量分析显示，转基因大鼠肾上腺11β-羟化酶mRNA略有增加，而在两种品系中大脑似乎表达几乎相同量的这种酶。细胞色素P45011B1 mRNA在不同细胞（可能是神经细胞）中，特别是在糖皮质激素受体高密度区域的表达，表明脑源性皮质酮在受体激活中可能具有功能。
