Elevated calcineurin activity in primary astrocytes leads to the dephosphorylation of connexin 43 in conjunction with increased membrane permeability.

IF 1.6 4区 医学 Q4 NEUROSCIENCES Neuroreport Pub Date : 2024-07-01 Epub Date: 2024-05-17 DOI:10.1097/WNR.0000000000002051
Blaine E Weiss, Susan D Kraner, Irina A Artiushin, Christopher M Norris
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Abstract

Hyperactivation of the Ca2+/calmodulin-dependent phosphatase calcineurin (CN) is observed in reactive astrocytes associated with neuroinflammation and progressive degenerative diseases, like Alzheimer's disease. Apart from key transcription factors (e.g. nuclear factor of activated t cells and nuclear factor-κB) very few other CN-dependent pathways have been studied in astrocytes. The hemichannel protein, connexin 43 (Cx43) is found at high levels in astrocytes and contains a CN-sensitive Ser residue near its carboxy terminus. CN-dependent dephosphorylation of Cx43 has been reported in primary astrocytes treated with injurious stimuli, but much remains unknown about CN/Cx43 interactions in the context of neuroinflammation and disease. Western blots were used to assess total Cx43 and dephosphorylated Cx43 subtypes in rat embryonic primary astrocytes treated with a hyperactive CN fragment (ΔCN, via adenovirus), or with a proinflammatory cytokine cocktail. Under similar treatment conditions, an ethidium bromide (EtBr) uptake assay was used to assess membrane permeability. Effects of ΔCN and cytokines were tested in the presence or absence of the CN inhibitor, cyclosporin A. A connexin inhibitor, carbenoxolone was also used in EtBr assays to assess the involvement of connexins in membrane permeability. Treatment with ΔCN or cytokines increased dephosphorylated Cx43 levels in conjunction with increased membrane permeability (elevated EtBr uptake). Effects of ΔCN or cytokine treatment were blocked by cyclosporine A. Treatment-induced changes in EtBr uptake were also inhibited by carbenoxolone. The results suggest that Cx43 hemichannels could be an important mechanism through which astrocytic CN disrupts neurologic function associated with neurodegenerative disease.

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原代星形胶质细胞中钙调素活性的升高会导致连接蛋白 43 的去磷酸化,同时增加膜的通透性。
在与神经炎症和渐进性退行性疾病(如阿尔茨海默病)相关的反应性星形胶质细胞中,可以观察到 Ca2+/calmodulin 依赖性磷酸酶钙调磷酸酶(CN)的过度激活。除了关键的转录因子(如活化 t 细胞核因子和核因子-κB)外,很少有人研究过星形胶质细胞中其他依赖 CN 的途径。在星形胶质细胞中发现了高水平的半通道蛋白--连接蛋白 43(Cx43),其羧基末端附近含有对 CN 敏感的 Ser 残基。据报道,在受到损伤性刺激的原代星形胶质细胞中,Cx43会发生CN依赖性去磷酸化,但在神经炎症和疾病的背景下,CN/Cx43之间的相互作用仍有很多未知之处。研究人员使用 Western 印迹来评估用高活性 CN 片段(ΔCN,通过腺病毒)或促炎细胞因子鸡尾酒处理的大鼠胚胎原代星形胶质细胞中的总 Cx43 和去磷酸化 Cx43 亚型。在类似的处理条件下,使用溴化乙锭(EtBr)摄取试验来评估膜通透性。在有或没有氯化萘抑制剂环孢素 A 的情况下,测试了ΔCN 和细胞因子的影响。用ΔCN或细胞因子处理会增加去磷酸化的Cx43水平,同时增加膜通透性(EtBr摄取量增加)。环孢素 A 可阻断ΔCN 或细胞因子处理的影响。这些结果表明,Cx43 半通道可能是星形胶质细胞 CN 干扰与神经退行性疾病相关的神经功能的重要机制。
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来源期刊
Neuroreport
Neuroreport 医学-神经科学
CiteScore
3.20
自引率
0.00%
发文量
150
审稿时长
1 months
期刊介绍: NeuroReport is a channel for rapid communication of new findings in neuroscience. It is a forum for the publication of short but complete reports of important studies that require very fast publication. Papers are accepted on the basis of the novelty of their finding, on their significance for neuroscience and on a clear need for rapid publication. Preliminary communications are not suitable for the Journal. Submitted articles undergo a preliminary review by the editor. Some articles may be returned to authors without further consideration. Those being considered for publication will undergo further assessment and peer-review by the editors and those invited to do so from a reviewer pool. The core interest of the Journal is on studies that cast light on how the brain (and the whole of the nervous system) works. We aim to give authors a decision on their submission within 2-5 weeks, and all accepted articles appear in the next issue to press.
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