Regulation and the Mechanism of Estrogen on Cav1.2 Gene in Rat-Cultured Cortical Astrocytes
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L-type calcium channel (LTCC) gene Cav1.2 is believed to play an important role in the alteration of Ca2+ homeostasis in brain astrocytes. Increasing evidence shows that alteration of intracellular Ca2+ concentration is related to the effect of 17β-estradiol (E2) in a variety of neurophysiological and neuropathological conditions. In this study, we measured immunoreactivity of Cav1.2 protein expression in rat primary cortical astrocytes by using Western blots. We demonstrated that E2 upregulated Cav1.2 expression in a dose- and time-dependent manner and the effect of E2 on Cav1.2 expression were blocked by an estrogen receptor (ER) antagonist, ICI-182,780. The ER subtype-selective ERα agonists propylpyrazole triole (PPT) and ERβ agonist diarylpropionitrile (DPN) both increase the expression of Cav1.2 in a dose-dependent manner. Also, the PPT most closely mimicked the upregulation of Cav1.2 protein expression by E2. Similar experiments of 10 nM E2-treated ERα- or ERβ-knockdown astrocytes have also shown that the E2 regulation of Cav1.2 protein expression is mediated through an ERα-dependent pathway. Furthermore, we established that E2 did not change the level of Cav1.2 mRNA. The induction of E2-mediated Cav1.2 expression was inhibited by cycloheximide (CHX) but not by actinomycin D (Act-D), suggesting that E2 regulation of Cav1.2 expression occurred at a posttranscriptional level. We also found that E2 may increase Cav1.2 levels by decreasing its ubiquitination and degradation rate. These findings provide new information about the effect of E2 on Cav1.2 in astrocytes, particularly necessary for the treatment of neurological disease.
Keywords17β-estradiol Astrocyte Cav1.2 Estrogen receptor
This work was supported by the National Natural Science Foundation of China (Nos. 81220108010). We would like to acknowledge the First Affiliated Hospital of Chongqing Medical University.
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflicts of interest.
All experimental procedures were performed in compliance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals, and all experimental protocols were approved by the Ethics Committee of Chongqing Medical University.
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