Cloning and characterization of the rat Slo3 (K(Ca)5.1) channel: From biophysics to pharmacology
Cloning and characterization of the rat Slo3 (K(Ca)5.1) channel: From biophysics to pharmacology
复制标题
大鼠 Slo3 (K(Ca)5.1) 通道的克隆和表征：从生物物理学到药理学
DOI：
10.1111/bph.15078
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发表时间：
2020
影响因子：
7.3
通讯作者：
Zhang Zhe
中科院分区：
文献类型：
--
作者：
Wang Guang-Ming;Zhong Zhi-Gang;Du Xiang-Rong;Zhang Fei-Fei;Guo Qing;Liu Ye;Tang Qiong-Yao;Zhang Zhe
背景与目的Slo 3钾（KCa5.1）通道在睾丸和精子中特异性表达，对于哺乳动物雄性受精至关重要。牛、小鼠和人Slo 3 α-亚基的序列差异揭示了不同物种之间的快速进化速率。大鼠Slo 3（rSlo 3）通道尚未被克隆和characterizedprevious.Experimental ApproachWe使用分子克隆，电生理学（inside‐out patches和outside‐out patches）和诱变来研究rSlo 3通道的生物物理性质和药理学特征。Key ResultsThe大鼠Slo 3通道（rSlo 3）是由电压和胞浆pH而不是细胞内钙门控的。rSlo 3通道的电压依赖性、pH敏感性和活化动力学特征与其他Slo 3直向同源物的特征不同。在药理学方面，rSlo 3通道的4-AP阻断也显示出与其mSlo 3通道阻断不同的性质。伊比利亚毒素和孕酮对rSlo 3通道有微弱的抑制作用。最后，我们发现，异丙酚，广泛使用的全身麻醉剂之一，从细胞内和细胞外的两侧阻断rSlo 3通道，而氯胺酮只阻断rSlo 3通道的细胞外side.Conclusion和ImplicationsOur研究结果表明，rSlo 3通道具有独特的生物物理和药理学特性。我们的研究结果为Slo 3家族通道的改变提供了新的见解，这对于估计使用这些药物改善精子质量的效果是有价值的。
Background and PurposeThe Slo3 potassium (KCa5.1) channel, which is specifically expressed in the testis and sperm, is essential for mammalian male fertilization. The sequence divergence of the bovine, mouse and human Slo3 α‐subunit revealed a rapid evolution rate across different species. The rat Slo3 (rSlo3) channel has not been cloned and characterized previously.Experimental ApproachWe used molecular cloning, electrophysiology (inside‐out patches and outside‐out patches) and mutagenesis to investigate the biophysical properties and pharmacological characteristics of the rSlo3 channel.Key ResultsThe rat Slo3 channel (rSlo3) is gated by voltage and cytosolic pH rather than intracellular calcium. The characteristics of voltage‐dependent, pH‐sensitivity and activation kinetics of the rSlo3 channel differ from the characteristics of other Slo3 orthologues. In terms of pharmacology, the 4‐AP blockade of the rSlo3 channel also shows properties distinct from its blockade of the mSlo3 channel. Iberiotoxin and progesterone weakly inhibit the rSlo3 channel. Finally, we found that propofol, one of the widely used general anaesthetics, blocks the rSlo3 channel from both intracellular and extracellular sides, whereas ketamine only blocks the rSlo3 channel at the extracellular side.Conclusion and ImplicationsOur findings suggest that the rSlo3 channel possesses unique biophysical and pharmacological properties. Our results provide new insights into the diversities of the Slo3 family of channels, which are valuable for estimating the effects of the use of these drugs to improve sperm quality.