Neha Gupta , Salah A. Baker , Kenton M. Sanders , Caoimhin S. Griffin , Gerard P. Sergeant , Mark A. Hollywood , Keith D. Thornbury , Bernard T. Drumm
{"title":"卡贾尔间质细胞样细胞(ICC-LC)表现出动态自发活动,但在小鼠尿道中没有功能神经支配","authors":"Neha Gupta , Salah A. Baker , Kenton M. Sanders , Caoimhin S. Griffin , Gerard P. Sergeant , Mark A. Hollywood , Keith D. Thornbury , Bernard T. Drumm","doi":"10.1016/j.ceca.2024.102931","DOIUrl":null,"url":null,"abstract":"<div><p>Urethral smooth muscle cells (USMC) contract to occlude the internal urethral sphincter during bladder filling. Interstitial cells also exist in urethral smooth muscles and are hypothesized to influence USMC behaviours and neural responses. These cells are similar to Kit<sup>+</sup> interstitial cells of Cajal (ICC), which are gastrointestinal pacemakers and neuroeffectors. Isolated urethral ICC-like cells (ICC-LC) exhibit spontaneous intracellular Ca<sup>2+</sup> signalling behaviours that suggest these cells may serve as pacemakers or neuromodulators similar to ICC in the gut, although observation and direct stimulation of ICC-LC within intact urethral tissues is lacking. We used mice with cell-specific expression of the Ca<sup>2+</sup> indicator, GCaMP6f, driven off the endogenous promoter for <em>Kit</em> (Kit-GCaMP6f mice) to identify ICC-LC <em>in situ</em> within urethra muscles and to characterize spontaneous and nerve-evoked Ca<sup>2+</sup> signalling. ICC-LC generated Ca<sup>2+</sup> waves spontaneously that propagated on average 40.1 ± 0.7 μm, with varying amplitudes, durations, and spatial spread. These events originated from multiple firing sites in cells and the activity between sites was not coordinated. ICC-LC in urethra formed clusters but not interconnected networks. No evidence for entrainment of Ca<sup>2+</sup> signalling between ICC-LC was obtained. Ca<sup>2+</sup> events in ICC-LC were unaffected by nifedipine but were abolished by cyclopiazonic acid and decreased by an antagonist of Orai Ca<sup>2+</sup> channels (GSK-7975A). Phenylephrine increased Ca<sup>2+</sup> event frequency but a nitric oxide donor (DEA-NONOate) had no effect. Electrical field stimulation (EFS, 10 Hz) of intrinsic nerves, which evoked contractions of urethral rings and increased Ca<sup>2+</sup> event firing in USMC, failed to evoke responses in ICC-LC. Our data suggest that urethral ICC-LC are spontaneously active but are not regulated by autonomic neurons.</p></div>","PeriodicalId":9678,"journal":{"name":"Cell calcium","volume":null,"pages":null},"PeriodicalIF":4.3000,"publicationDate":"2024-07-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0143416024000897/pdfft?md5=b7812ffdecf9e29cc0881b72c750836c&pid=1-s2.0-S0143416024000897-main.pdf","citationCount":"0","resultStr":"{\"title\":\"Interstitial cell of Cajal-like cells (ICC-LC) exhibit dynamic spontaneous activity but are not functionally innervated in mouse urethra\",\"authors\":\"Neha Gupta , Salah A. Baker , Kenton M. Sanders , Caoimhin S. Griffin , Gerard P. Sergeant , Mark A. Hollywood , Keith D. Thornbury , Bernard T. Drumm\",\"doi\":\"10.1016/j.ceca.2024.102931\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Urethral smooth muscle cells (USMC) contract to occlude the internal urethral sphincter during bladder filling. Interstitial cells also exist in urethral smooth muscles and are hypothesized to influence USMC behaviours and neural responses. These cells are similar to Kit<sup>+</sup> interstitial cells of Cajal (ICC), which are gastrointestinal pacemakers and neuroeffectors. Isolated urethral ICC-like cells (ICC-LC) exhibit spontaneous intracellular Ca<sup>2+</sup> signalling behaviours that suggest these cells may serve as pacemakers or neuromodulators similar to ICC in the gut, although observation and direct stimulation of ICC-LC within intact urethral tissues is lacking. We used mice with cell-specific expression of the Ca<sup>2+</sup> indicator, GCaMP6f, driven off the endogenous promoter for <em>Kit</em> (Kit-GCaMP6f mice) to identify ICC-LC <em>in situ</em> within urethra muscles and to characterize spontaneous and nerve-evoked Ca<sup>2+</sup> signalling. ICC-LC generated Ca<sup>2+</sup> waves spontaneously that propagated on average 40.1 ± 0.7 μm, with varying amplitudes, durations, and spatial spread. These events originated from multiple firing sites in cells and the activity between sites was not coordinated. ICC-LC in urethra formed clusters but not interconnected networks. No evidence for entrainment of Ca<sup>2+</sup> signalling between ICC-LC was obtained. Ca<sup>2+</sup> events in ICC-LC were unaffected by nifedipine but were abolished by cyclopiazonic acid and decreased by an antagonist of Orai Ca<sup>2+</sup> channels (GSK-7975A). Phenylephrine increased Ca<sup>2+</sup> event frequency but a nitric oxide donor (DEA-NONOate) had no effect. Electrical field stimulation (EFS, 10 Hz) of intrinsic nerves, which evoked contractions of urethral rings and increased Ca<sup>2+</sup> event firing in USMC, failed to evoke responses in ICC-LC. 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引用次数: 0
摘要
尿道平滑肌细胞(USMC)会在膀胱充盈时收缩以闭合尿道内括约肌。尿道平滑肌中也存在间质细胞,据推测,它们会影响尿道平滑肌细胞的行为和神经反应。这些细胞类似于 Kit interstitial cells of Cajal (ICC),后者是胃肠道起搏器和神经效应器。分离的尿道 ICC 样细胞(ICC-LC)表现出自发的细胞内 Ca 信号行为,这表明这些细胞可能充当类似于肠道 ICC 的起搏器或神经调节器,尽管目前还缺乏对完整尿道组织内 ICC-LC 的观察和直接刺激。我们使用细胞特异性表达钙离子指示剂 GCaMP6f 的小鼠(Kit-GCaMP6f 小鼠)来鉴定尿道肌肉中的 ICC-LC,并描述自发和神经诱发的钙离子信号。ICC-LC 自发产生的 Ca 波平均传播 40.1 ± 0.7 μm,振幅、持续时间和空间扩散各不相同。这些事件源于细胞中的多个点燃点,而点燃点之间的活动并不协调。尿道中的 ICC-LC 形成了集群,但没有形成相互连接的网络。没有证据表明ICC-LC之间的钙离子信号是相互关联的。ICC-LC 中的 Ca 事件不受硝苯地平的影响,但会被环噻唑啉酸所取消,并因 Orai Ca 通道拮抗剂(GSK-7975A)而减少。苯肾上腺素增加了钙离子事件频率,但一氧化氮供体(DEA-NONOate)没有影响。对固有神经的电场刺激(EFS,10 Hz)可诱发尿道环收缩并增加 USMC 的 Ca 事件发射,但未能诱发 ICC-LC 的反应。我们的数据表明,尿道 ICC-LC 自发活跃,但不受自律神经元的调节。
Interstitial cell of Cajal-like cells (ICC-LC) exhibit dynamic spontaneous activity but are not functionally innervated in mouse urethra
Urethral smooth muscle cells (USMC) contract to occlude the internal urethral sphincter during bladder filling. Interstitial cells also exist in urethral smooth muscles and are hypothesized to influence USMC behaviours and neural responses. These cells are similar to Kit+ interstitial cells of Cajal (ICC), which are gastrointestinal pacemakers and neuroeffectors. Isolated urethral ICC-like cells (ICC-LC) exhibit spontaneous intracellular Ca2+ signalling behaviours that suggest these cells may serve as pacemakers or neuromodulators similar to ICC in the gut, although observation and direct stimulation of ICC-LC within intact urethral tissues is lacking. We used mice with cell-specific expression of the Ca2+ indicator, GCaMP6f, driven off the endogenous promoter for Kit (Kit-GCaMP6f mice) to identify ICC-LC in situ within urethra muscles and to characterize spontaneous and nerve-evoked Ca2+ signalling. ICC-LC generated Ca2+ waves spontaneously that propagated on average 40.1 ± 0.7 μm, with varying amplitudes, durations, and spatial spread. These events originated from multiple firing sites in cells and the activity between sites was not coordinated. ICC-LC in urethra formed clusters but not interconnected networks. No evidence for entrainment of Ca2+ signalling between ICC-LC was obtained. Ca2+ events in ICC-LC were unaffected by nifedipine but were abolished by cyclopiazonic acid and decreased by an antagonist of Orai Ca2+ channels (GSK-7975A). Phenylephrine increased Ca2+ event frequency but a nitric oxide donor (DEA-NONOate) had no effect. Electrical field stimulation (EFS, 10 Hz) of intrinsic nerves, which evoked contractions of urethral rings and increased Ca2+ event firing in USMC, failed to evoke responses in ICC-LC. Our data suggest that urethral ICC-LC are spontaneously active but are not regulated by autonomic neurons.
期刊介绍:
Cell Calcium covers the field of calcium metabolism and signalling in living systems, from aspects including inorganic chemistry, physiology, molecular biology and pathology. Topic themes include:
Roles of calcium in regulating cellular events such as apoptosis, necrosis and organelle remodelling
Influence of calcium regulation in affecting health and disease outcomes