中国神经再生研究(英文版) ›› 2022, Vol. 17 ›› Issue (11): 2465-2471.doi: 10.4103/1673-5374.339008

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

孕酮对浦肯野细胞中T型Ca2+通道表达的影响

  

  • 出版日期:2022-11-15 发布日期:2022-04-22

Effects of progesterone on T-type-Ca2+-channel expression in Purkinje cells

Annika Eickhoff1, Jonas Tjaden1, Sarah Stahlke1, Matthias Vorgerd2, Verena Theis1, Veronika Matschke1, #, Carsten Theiss1, *, #   

  1. 1Department of Cytology, Institute of Anatomy, Ruhr-University Bochum, Bochum, Germany; 2Department of Neurology, Neuromuscular Center Ruhrgebiet, University Hospital Bergmannsheil, Ruhr-Universität Bochum, Bochum, Germany
  • Online:2022-11-15 Published:2022-04-22
  • Contact: Carsten Theiss, Dr. rer. nat., carsten.theiss@rub.de.

摘要:

孕酮通过孕酮受体A和B刺激小脑浦肯野细胞树突、树突棘、突触形成影响神经元可塑性,但孕酮对神经可塑性的进一步影响尚在研究中。实验应用浦肯野细胞培养物分析孕酮对电压门控T型Ca2+通道Cav3.1、Cav3.2和Cav3.3的分子水平的影响。结果显示,黄体酮对T型Ca2+通道的mRNA表达有直接影响,也对参与下游信号通路的蛋白激酶A和C有直接影响,这些信号通路在神经元可塑性中发挥着重要作用。使用激光显微切割和不同成熟阶段的纯化浦肯野细胞培养物检测T型Ca2+通道和信号级联的蛋白激酶的mRNA表达。以免疫组织化学染色确定T型Ca2+通道在浦肯野细胞中的定位特征。对纯化的浦肯野细胞培养物进行24和48h孕酮处理的结果表明,孕酮刺激并增加了浦肯野细胞中Cav3.1和Cav3.3以及相关蛋白激酶A和C的mRNA水平的表达。这些作用扩展了目前对孕酮在中枢神经系统功能的认识,并为其对神经可塑性的影响提供了一种新的解释。

https://orcid.org/0000-0001-7983-0143 (Carsten Theiss)

Abstract: Plasticity of cerebellar Purkinje cells (PC) is influenced by progesterone via the classical progesterone receptors PR-A and PR-B by stimulating dendritogenesis, spinogenesis, and synaptogenesis in these cells. Dissociated PC cultures were used to analyze progesterone effects at a molecular level on the voltage-gated T-type-Ca2+-channels Cav3.1, Cav3.2, and Cav3.3 as they helped determine neuronal plasticity by regulating Ca2+-influx in neuronal cells. The results showed direct effects of progesterone on the mRNA expression of T-type-Ca2+-channels, as well as on the protein kinases A and C being involved in downstream signaling pathways that play an important role in neuronal plasticity. For the mRNA expression studies of T-type-Ca2+-channels and protein kinases of the signaling cascade, laser microdissection and purified PC cultures of different maturation stages were used. Immunohistochemical staining was also performed to characterize the localization of T-type-Ca2+-channels in PC. Experimental progesterone treatment was performed on the purified PC culture for 24 and 48 hours. Our results show that progesterone increases the expression of Cav3.1 and Cav3.3 and associated protein kinases A and C in PC at the mRNA level within 48 hours after treatment at latest. These effects extend the current knowledge of the function of progesterone in the central nervous system and provide an explanatory approach for its influence on neuronal plasticity.

Key words: Cav3.1, Cav3.2, Cav3.3, neuronal plasticity, progesterone, protein kinase A, protein kinase C, Purkinje cells, rat cerebellum, T-type-Ca2+-channels