Expression and localization of 25-Dx, a membrane-associated putative progesterone-binding protein, in the developing Purkinje cell
Expression and localization of 25-Dx, a membrane-associated putative progesterone-binding protein, in the developing Purkinje cell
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DOI：
10.1016/j.neuroscience.2004.04.003
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发表时间：
2004-01-01
期刊：
影响因子：
3.3
通讯作者：
Tsutsui, K
中科院分区：
文献类型：
--
作者：
Sakamoto, H;Ukena, K;Tsutsui, K
神经类固醇在大脑中从头合成，小脑浦肯野细胞是神经类固醇形成的主要部位。我们已经证明，大鼠浦肯野细胞仅在新生儿时期就能从胆固醇中主动产生孕酮，孕酮通过其在该神经元中的核受体促进树突生长、脊柱发生和突触发生。另一方面，25-Dx，一种假定的膜孕酮受体，已经在大鼠肝脏中被鉴定出来。因此，在本研究中，我们研究了25-Dx在浦肯野细胞中的表达和定位，以了解黄体酮在该神经元中的作用模式。逆转录pcr和Western免疫印迹分析显示，25-Dx mRNA和25-Dx样蛋白在大鼠小脑中表达，在新生期增加。免疫细胞化学证实25- dx样蛋白的表达定位于浦肯野细胞和外颗粒细胞层。在超微结构水平上，我们进一步发现25- dx样免疫反应性与浦肯野细胞内质网和高尔基体的膜结构有关。这些结果表明浦肯野细胞在新生儿时期表达了假定的膜孕酮受体25-Dx。孕酮可能通过25-Dx及其在新生儿浦肯野细胞中的核受体促进树突生长、脊髓发生和突触发生。(c) 2004 ibro。Elsevier Ltd.出版。版权所有。
Neurosteroids are synthesized de novo in the brain and the cerebellar Purkinje cell is a major site for neurosteroid formation. We have demonstrated that the rat Purkinje cell actively produces progesterone de novo from cholesterol only during neonatal life and progesterone promotes dendritic growth, spinogenesis and synaptogenesis via its nuclear receptor in this neuron. On the other hand, 25-Dx, a putative membrane progesterone receptor, has been identified in the rat liver. In this study, we therefore investigated the expression and localization of 25-Dx in the Purkinje cell to understand the mode of progesterone actions in this neuron. Reverse transcription-PCR and Western immunoblot analyses revealed the expressions of 25-Dx mRNA and 25-Dx-like protein in the rat cerebellum, which increased during neonatal life. By immunocytochemistry, the expression of 25-Dx-like protein was localized in the Purkinje cell and external granule cell layer. At the ultrastructural level, we further found that 25-Dx-like immunoreactivity was associated with membrane structures of the endoplasmic reticulum and Golgi apparatus in the Purkinje cell. These results indicate that the Purkinje cell expresses the putative membrane progesterone receptor, 25-Dx during neonatal life. Progesterone may promote dendritic growth, spinogenesis and synaptogenesis via 25-Dx as well as its nuclear receptor in the Purkinje cell in the neonate. (C) 2004 IBRO. Published by Elsevier Ltd. All rights reserved.