Gupta Neha, Baker Salah A, Sanders Kenton M, Rabab Kaneez E, Thean Denzel Kf, Alkawadri Tuleen, Griffin Caoimhin S, Sergeant Gerard P, Hollywood Mark A, Thornbury Keith D, Drumm Bernard T
Smooth Muscle Research Centre, Department of Life & Health Science, Dundalk Institute of Technology, Dundalk, Co. Louth, Ireland.
Department of Physiology & Cell Biology, University of Nevada, Reno School of Medicine, Reno, NV, USA.
Sci Rep. 2025 May 19;15(1):17365. doi: 10.1038/s41598-025-00953-z.
Anoctamin-1 Ca-activated Cl channels (ANO1) are proposed to modulate contractility of urethra smooth muscle cells (USMC), but their cellular expression and contribution to agonist/neural evoked activity is unclear. ANO1 is implicated as a potential target to treat incontinence, thus this is an important issue to resolve. We sought to clarify roles of ANO1 in contractility of mouse USMC. We found expression of Ano1 transcripts in murine urethra, with no difference between male and females. Immunolabelling revealed ANO1 was expressed in USMC and not in specialized populations of interstitial cells (c-kit interstitial of Cajal-like cells (ICC-LC) and PDGFRα cells). However, a specific ANO1 channel inhibitor, Ani9, failed to affect urethral contractions elicited by phenylephrine, arginine vasopressin or electrical field stimulation of intrinsic nerves. CaCCA01 also failed to affect urethral contractions. In addition, Ani9 had no effect on Ca signals generated by USMC in situ. In contrast, Ani9 effectively reduced spontaneous contractions and Ca signals of mouse proximal colon. In addition, Ani9 inhibited ANO1 currents recorded in HEK 293 cells, at concentrations 30 times less than those used in organ bath experiments. Our data suggest that despite expression of ANO1 in USMC, these channels do not contribute to basal Ca signalling, or agonist and neurally-evoked contractions in murine urethra.
据推测，anoctamin-1钙激活氯离子通道（ANO1）可调节尿道平滑肌细胞（USMC）的收缩性，但其细胞表达以及对激动剂/神经诱发活动的作用尚不清楚。ANO1被认为是治疗尿失禁的潜在靶点，因此这是一个亟待解决的重要问题。我们试图阐明ANO1在小鼠USMC收缩性中的作用。我们发现小鼠尿道中存在Ano1转录本的表达，雄性和雌性之间无差异。免疫标记显示ANO1在USMC中表达，而在特殊的间质细胞群体（类Cajal间质细胞（ICC-LC）和血小板衍生生长因子受体α（PDGFRα）细胞）中不表达。然而，一种特异性ANO1通道抑制剂Ani9未能影响去氧肾上腺素、精氨酸加压素或内在神经的电场刺激所引发的尿道收缩。CaCCA01也未能影响尿道收缩。此外，Ani9对原位USMC产生的钙信号没有影响。相比之下，Ani9能有效降低小鼠近端结肠的自发收缩和钙信号。此外，在浓度比器官浴实验中所用浓度低30倍的情况下，Ani9就能抑制在人胚肾293（HEK 293）细胞中记录到的ANO1电流。我们的数据表明，尽管ANO1在USMC中表达，但这些通道对小鼠尿道的基础钙信号、激动剂诱发的收缩和神经诱发的收缩均无作用。
