Martínez Ana Alonso, Muñoz Yolanda Ruiz, Serrano Fuencisla San Juan, García Pilar Molist
Department of Functional Biology and Health Sciences, University of Vigo, 36310 Vigo, Spain.
J Comp Physiol B. 2008 Jul;178(5):647-54. doi: 10.1007/s00360-008-0256-x. Epub 2008 Mar 14.
We have examined the localization of P450scc in different tissues of the mollusc Mytilus galloprovincialis along a gonadal cycle by using a polyclonal antibody against rat cholesterol side-chain cleavage enzyme (cytochrome P450scc). Immunoreactivity specific for P450scc was found only in the cytoplasm of basophilic cells from digestive gland; gonad, gills and kidney appeared to be devoid of P450scc immunoreactivity. SDS-gel electrophoresis and western blot analysis of different subcellular fractions revealed that this protein is mainly located in microsomes, has a molecular weight of 51.4 kDa and is recognized by the antibody against rat P450scc. These results suggest that the digestive gland of M. galloprovincialis could be considered a steroidogenic tissue where the first step in biosynthesis of steroid hormones occurs. Moreover, seasonal slot blot analysis of post-mitochondrial fractions showed a P450scc induction by available nutrients and phytoplankton blooms.
我们使用针对大鼠胆固醇侧链裂解酶(细胞色素P450scc)的多克隆抗体,研究了在性腺周期中,P450scc在软体动物地中海贻贝不同组织中的定位。仅在消化腺嗜碱性细胞的细胞质中发现了对P450scc具有特异性的免疫反应性;性腺、鳃和肾脏似乎没有P450scc免疫反应性。对不同亚细胞组分进行的SDS凝胶电泳和蛋白质免疫印迹分析表明,该蛋白主要位于微粒体中,分子量为51.4 kDa,并且能被抗大鼠P450scc的抗体识别。这些结果表明,地中海贻贝的消化腺可被视为一个类固醇生成组织,类固醇激素生物合成的第一步在此发生。此外,对线粒体后组分的季节性狭缝印迹分析表明,可利用的营养物质和浮游植物大量繁殖会诱导P450scc的产生。