Bijos Dominika A, Drake Marcus J, Vahabi Bahareh
School of Clinical Sciences, University of Bristol, Bristol, United Kingdom; Bristol Urological Institute, North Bristol NHS Trust, Bristol, United Kingdom.
Bristol Urological Institute, North Bristol NHS Trust, Bristol, United Kingdom; Department of Biological, Biomedical and Analytical Sciences, University of the West of England, Bristol, United Kingdom.
PLoS One. 2014 Sep 2;9(9):e106190. doi: 10.1371/journal.pone.0106190. eCollection 2014.
To investigate presence, location and functional role of calcium-activated chloride channel (CaCC) Anoctamin-1 (Ano1) in rat urinary bladder.
Bladders from 3 week old Wistar rats were studied. End-point PCR on total mRNA was used to assess the expression of Ano1. Immunofluorescent labelling of whole mount bladder tissue imaged with confocal microscope allowed localization of Ano1 and vimentin immunopositive cells. The effects of CaCC blockers: niflumic acid (NFA) (3,10,30 µM) and 5-Nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) (10, 30 µM) on spontaneous phasic contractile activity of intact (with mucosa) and denuded (without mucosa) detrusor strips were measured under isometric tension in organ baths (n = 141, N = 60).
Ano1 expression was found at mRNA level in mucosa and detrusor layers. Confocal microscopy revealed presence of Ano1 immunopositive cells in mucosa and in detrusor layers; a subpopulation of vimentin positive cells expressed Ano1. Both chloride channel blockers reduced the amplitude and frequency of phasic contractions in denuded and intact strips.
Ano1 is expressed in rat urinary bladder and is present in cells sharing markers with interstitial cells. CaCC blockers reduced phasic activity of the bladder tissue. Ano1 is expressed in the bladder and plays a role in its spontaneous phasic contractile activity.
研究钙激活氯离子通道（CaCC）无翅型MMTV整合位点家族成员1（Ano1）在大鼠膀胱中的存在情况、位置及功能作用。
研究3周龄Wistar大鼠的膀胱。采用总mRNA的终点PCR法评估Ano1的表达。用共聚焦显微镜对膀胱全层组织进行免疫荧光标记成像，以定位Ano1和波形蛋白免疫阳性细胞。在器官浴槽中，在等长张力条件下，测量CaCC阻滞剂：尼氟灭酸（NFA）（3、10、30 μM）和5-硝基-2-（3-苯丙基氨基）苯甲酸（NPPB）（10、30 μM）对完整（有黏膜）和去黏膜（无黏膜）逼尿肌条自发相性收缩活动的影响（n = 141，N = 60）。
在黏膜和逼尿肌层的mRNA水平发现了Ano1表达。共聚焦显微镜显示黏膜和逼尿肌层存在Ano1免疫阳性细胞；波形蛋白阳性细胞亚群表达Ano1。两种氯离子通道阻滞剂均降低了去黏膜和完整肌条的相性收缩幅度和频率。
Ano1在大鼠膀胱中表达，且存在于与间质细胞有共同标志物的细胞中。CaCC阻滞剂降低了膀胱组织的相性活动。Ano1在膀胱中表达，并在其自发相性收缩活动中起作用。
