Generation of Kcnma1fl-tdTomato, a conditional deletion of the BK channel α subunit in mouse.
Generation of Kcnma1fl-tdTomato, a conditional deletion of the BK channel α subunit in mouse.
复制标题
DOI：
10.14814/phy2.12612
复制
发表时间：
2015-11
影响因子：
2.5
通讯作者：
Meredith AL
中科院分区：
文献类型：
--
作者：
Zemen BG;Lai MH;Whitt JP;Khan Z;Zhao G;Meredith AL
BK 大电导钙激活 K+ 通道 (KCa1.1) 在许多组织中广泛表达，有助于心血管、神经和其他特殊生理功能的系统调节。成孔α亚基由Kcnma1基因编码，最初在小鼠中被命名为mSlo1，在果蝇中被命名为slowpoke。小鼠（Kcnma1−/−）的整体缺失会导致神经元和肌肉兴奋性以及与非兴奋性细胞通道功能相关的其他表型出现大量缺陷。虽然纯合子无效小鼠是可行的，但 BK 功能的普遍丧失使得对涉及独立表达 BK 通道的多种细胞类型相互作用的表型的解释变得复杂。在这里，我们报告了使用 Cre-loxP 系统 (Kcnma1fl-tdTomato) 生成用于 Kcnma1 条件失活的目标等位基因。 Cre 介导的重组产生无效等位基因，并且在来自 Nestin-Cre 的神经元和肌肉细胞中检测不到 BK 电流； Kcnma1fl/fl 和 SM22α-Cre；分别为 Kcnma1fl/fl 小鼠。在 Cre 表达组织中检测到 tdTomato 表达，但在 Cre 阴性对照中未检测到。这些数据证明了 Kcnma1fl-tdTomato 在条件性删除 BK 通道方面的实用性，有助于了解组织特异性对体内生理功能的贡献。
BK large conductance calcium-activated K+ channels (KCa1.1) are expressed widely across many tissues, contributing to systemic regulation of cardiovascular, neurological, and other specialized physiological functions. The pore-forming α subunit is encoded by the Kcnma1 gene, originally named mSlo1 in mouse and slowpoke in Drosophila. Global deletion in mouse (Kcnma1−/−) produces a plethora of defects in neuron and muscle excitability, as well as other phenotypes related to channel function in nonexcitable cells. While homozygous null mice are viable, the ubiquitous loss of BK function has complicated the interpretation of phenotypes involving the interaction of multiple cell types which independently express BK channels. Here, we report the generation of a targeted allele for conditional inactivation of Kcnma1 using the Cre-loxP system (Kcnma1fl-tdTomato). Cre-mediated recombination generates a null allele, and BK currents were not detectable in neurons and muscle cells from Nestin-Cre; Kcnma1fl/fl and SM22α-Cre; Kcnma1fl/fl mice, respectively. tdTomato expression was detected in Cre-expressing tissues, but not in Cre-negative controls. These data demonstrate the utility of Kcnma1fl-tdTomato for conditional deletion of the BK channel, facilitating the understanding of tissue-specific contributions to physiological function in vivo.