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首页> 外文期刊>BMC Physiology >Differential role of STIM1 and STIM2 during transient inward (T in) current generation and the maturation process in the Xenopus oocyte
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Differential role of STIM1 and STIM2 during transient inward (T in) current generation and the maturation process in the Xenopus oocyte

机译：STIM1和STIM2在非洲爪蟾卵母细胞瞬时内向（T in）电流生成和成熟过程中的差异作用

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Background The Xenopus oocyte is a useful cell model to study Ca2+ homeostasis and cell cycle regulation, two highly interrelated processes. Here, we used antisense oligonucleotides to investigate the role in the oocyte of stromal interaction molecule (STIM) proteins that are fundamental elements of the store-operated calcium-entry (SOCE) phenomenon, as they are both sensors for Ca2+ concentration in the intracellular reservoirs as well as activators of the membrane channels that allow Ca2+ influx. Results Endogenous STIM1 and STIM2 expression was demonstrated, and their synthesis was knocked down 48–72?h after injecting oocytes with specific antisense sequences. Selective elimination of their mRNA and protein expression was confirmed by PCR and Western blot analysis, and we then evaluated the effect of their absence on two endogenous responses: the opening of SOC channels elicited by G protein-coupled receptor (GPCR)-activated Ca2+ release, and the process of maturation stimulated by progesterone. Activation of SOC channels was monitored electrically by measuring the Tin response, a Ca2+-influx-dependent Cl? current, while maturation was assessed by germinal vesicle breakdown (GVBD) scoring and electrophysiology. Conclusions It was found that STIM2, but not STIM1, was essential in both responses, and Tin currents and GVBD were strongly reduced or eliminated in cells devoid of STIM2; STIM1 knockdown had no effect on the maturation process, but it reduced the Tin response by 15 to 70%. Thus, the endogenous SOCE response in Xenopus oocytes depended mainly on STIM2, and its expression was necessary for entry into meiosis induced by progesterone.
机译：背景爪蟾卵母细胞是研究Ca 2 + 稳态和细胞周期调控这两个高度相关的过程的有用细胞模型。在这里，我们使用反义寡核苷酸来研究基质相互作用分子（STIM）蛋白在卵母细胞中的作用，这些蛋白是储存操作性钙进入（SOCE）现象的基本要素，因为它们都是Ca 2+的传感器细胞内储库中的浓度以及允许Ca 2 + 流入的膜通道激活剂。结果证实了内源性STIM1和STIM2的表达，并且在向卵母细胞中注入特定的反义序列后，它们的合成被敲低了48–72？h。通过PCR和Western印迹分析证实了其mRNA和蛋白表达的选择性消除，然后我们评估了它们的缺失对两种内源性应答的影响：G蛋白偶联受体（GPCR）激活的Ca < sup> 2 + 释放，以及孕酮刺激的成熟过程。通过测量T in 响应，Ca 2 + -流入依赖的Cl ？电流来电气监测SOC通道的激活，通过生小泡破坏（GVBD）评分和电生理评估。结论发现，STIM2而不是STIM1在两种反应中都是必不可少的，并且在缺乏STIM2的细胞中T in 电流和GVBD被强烈降低或消除。 STIM1敲低对成熟过程没有影响，但将T in 响应降低了15％至70％。因此，非洲爪蟾卵母细胞中的内源性SOCE反应主要取决于STIM2，其表达对于进入孕激素诱导的减数分裂是必需的。

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