Discipline of Human Physiology, Center for Neuroscience, School of Medicine, Flinders University Adelaide, SA, Australia ; Department of Physiology & Cell Biology, University of Nevada School of Medicine Reno, NV, USA.
Front Neurosci. 2013 Oct 14;7:184. doi: 10.3389/fnins.2013.00184. eCollection 2013.
It is well established that the intrinsic pacemaker mechanism that generates cyclical colonic migrating motor complexes (CMMCs) does not require endogenous nitric oxide (NO). However, pharmacological blockade of endogenous NO production potently increases the frequency of CMMCs, suggesting that endogenous NO acts normally to inhibit the CMMC pacemaker mechanism. In this study, we investigated whether mice with a life long genetic deletion of the neuronal nitric oxide synthase (nNOS) gene would show similar CMMC characteristics as wild type mice that have endogenous NO production acutely inhibited. Intracellular electrophysiological and mechanical recordings were made from circular muscle cells of isolated whole mouse colon in wild type and nNOS knockout (KO) mice at 35°C. In wild type mice, the NOS inhibitor, L-NA (100 μM) caused a significant increase in CMMC frequency and a significant depolarization of the CM layer. However, unexpectedly, the frequency of CMMCs in nNOS KO mice was not significantly different from control mice. Also, the resting membrane potential of CM cells in nNOS KO mice was not depolarized compared to controls; and the amplitude of the slow depolarization phase underlying MCs was of similar amplitude between KO and wild type offspring. These findings show that in nNOS KO mice, the major characteristics of CMMCs and their electrical correlates are, at least in adult mice, indistinguishable from wild type control offspring. One possibility why the major characteristics of CMMCs were no different between both types of mice is that nNOS KO mice may compensate for their life long deletion of the nNOS gene, and their permanent loss of neuronal NO production. In this regard, we suggest caution should be exercised when assuming that data obtained from adult nNOS KO mice can be directly extrapolated to wild type mice, that have been acutely exposed to an inhibitor of NOS.
众所周知,产生周期性结肠移行性运动复合波(CMMC)的内在起搏机制不需要内源性一氧化氮(NO)。然而,内源性 NO 产生的药理学阻断强烈增加了 CMMC 的频率,这表明内源性 NO 正常作用以抑制 CMMC 起搏机制。在这项研究中,我们研究了具有终身基因缺失神经元型一氧化氮合酶(nNOS)的小鼠是否会表现出与具有急性抑制内源性 NO 产生的野生型小鼠相似的 CMMC 特征。在 35°C 下,从野生型和 nNOS 敲除(KO)小鼠的分离全鼠结肠的环形肌细胞中进行细胞内电生理和机械记录。在野生型小鼠中,NOS 抑制剂 L-NA(100 μM)导致 CMMC 频率显着增加和 CM 层显着去极化。然而,出乎意料的是,nNOS KO 小鼠的 CMMC 频率与对照小鼠没有显着差异。此外,与对照相比,nNOS KO 小鼠 CM 细胞的静息膜电位没有去极化;并且 MC 下的慢去极化相的幅度在 KO 和野生型后代之间相似。这些发现表明,在 nNOS KO 小鼠中,CMMC 的主要特征及其电相关性,至少在成年小鼠中,与野生型对照后代没有区别。为什么两种类型的小鼠之间的 CMMC 的主要特征没有差异的一种可能性是,nNOS KO 小鼠可能会补偿其终身缺失的 nNOS 基因及其神经元 NO 产生的永久性丧失。在这方面,我们建议在假设从成年 nNOS KO 小鼠获得的数据可以直接外推到已经急性暴露于 NOS 抑制剂的野生型小鼠时应谨慎行事。