Huang Zitong, Liao Lu, Wang Zhesheng, Lu Yulin, Yan Weiming, Cao Hongying, Tan Bo
Research Center for Integrative Medicine, School of Basic Medical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, PR China.
The Third Clinical Medical College, Guangzhou University of Chinese Medicine, Guangzhou, PR China.
J Neurosci Methods. 2021 Jan 15;348:109012. doi: 10.1016/j.jneumeth.2020.109012. Epub 2020 Nov 26.
The nerve plexus of the enteric nervous system (ENS) plays a crucial part in regulating gastrointestinal functions, such as muscle contractile activity and nutrient absorption. Studying the nerve plexus can provide vital information for research on ENS disorders. Whole-mount preparation is an important technique for investigating the nerve plexus. However, currently available methods are time consuming and highly technical.
This study describes a simple and rapid method for preparing whole mounts of the longitudinal muscle and myenteric plexuses (LMMPs) of rat colon. Integral LMMPs can be easily separated from the underlying layer by using glass rods and wet cotton swabs.
The proposed method allows the easy separation of the LMMPs in intact sheets. Immunofluorescence histochemical staining of whole mounts enable clear visualization of enteric ganglia, nerve fibers, and several subtypes of neuronal populations residing in the myenteric plexus.
Compared with existing procedures for whole-mount preparations, the proposed method achieves a quicker and more efficient preparation of high-quality LMMPs from intestinal segments in sufficient quantity.
This work provides a rapid method for efficiently preparing whole mounts of the intestines. Our method can be used for in situ observation of the morphological and functional alterations of the myenteric plexus for further studies on the ENS.
肠神经系统(ENS)的神经丛在调节胃肠功能(如肌肉收缩活动和营养吸收)中起着关键作用。研究神经丛可为ENS疾病的研究提供重要信息。整装标本制备是研究神经丛的一项重要技术。然而,目前可用的方法既耗时又技术要求高。
本研究描述了一种简单快速的制备大鼠结肠纵肌和肌间神经丛(LMMPs)整装标本的方法。使用玻璃棒和湿棉签可轻松将完整的LMMPs与下层分离。
所提出的方法可轻松分离出完整薄片中的LMMPs。整装标本的免疫荧光组织化学染色能够清晰地观察到肠神经节、神经纤维以及肌间神经丛中存在的几种神经元群体亚型。
与现有的整装标本制备程序相比,所提出的方法能更快速、高效地从肠段制备出足够数量的高质量LMMPs。
本研究提供了一种快速有效制备肠整装标本的方法。我们的方法可用于原位观察肌间神经丛的形态和功能变化,以进一步研究ENS。