4.4 Article

An efficient approach for wholemount preparation of the myenteric plexus of rat colon

Journal

JOURNAL OF NEUROSCIENCE METHODS
Volume 348, Issue -, Pages -

Publisher

ELSEVIER
DOI: 10.1016/j.jneumeth.2020.109012

Keywords

Enteric nervous system; Myenteric plexus; Separation; Whole mount

Funding

  1. National Natural Science Foundation of China [81573715, 81973586]

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This study presents a simple and rapid method for preparing whole mounts of rat colon, facilitating the easy separation of LMMPs and allowing clear visualization of structures through immunofluorescence staining. Compared to existing methods, this approach is more efficient and provides sufficient high-quality LMMPs in a quicker manner.
Background: 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. New method: 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. Results: 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. Comparison with existing methods: 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. Conclusions: 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.

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