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利用转运时间、组织采集和全层免疫染色对小鼠纵向肠道运动进行表征的方案。

Protocol to characterize longitudinal gut motility in mice using transit time, tissue harvest, and whole-mount immunostaining.

作者信息

Frith Mary E, Kashyap Purna C, Linden David R, Chang Eugene B

机构信息

Interdisciplinary Scientist Training Program, University of Chicago, Chicago, IL 60637, USA; Department of Medicine, University of Chicago, Chicago, IL 60637, USA.

Division of Gastroenterology and Hepatology, Department of Medicine, Mayo Clinic, Rochester, MN 55905, USA; Enteric Neuroscience Program, Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, MN 55905, USA.

出版信息

STAR Protoc. 2025 Jun 20;6(2):103761. doi: 10.1016/j.xpro.2025.103761. Epub 2025 Apr 15.

DOI:10.1016/j.xpro.2025.103761
PMID:40238633
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12022682/
Abstract

Transit time is a key in vivo metric of gastrointestinal (GI) motility, which is a physiologic readout of cellular communication within the enteric system. Here, we present a protocol to characterize longitudinal gut motility in mice. We describe steps for transit testing, whole-mount immunostaining, and tissue harvest. We then detail procedures for image processing and manual cell counting. This protocol seeks to minimize inter-trial variability while assessing cellular and molecular features that may underpin motility differences between experimental conditions. For complete details on the use and execution of this protocol, please refer to Frith et al..

摘要

转运时间是胃肠道（GI）蠕动的一项关键体内指标，而胃肠道蠕动是肠神经系统内细胞通讯的生理读数。在此，我们展示了一种用于表征小鼠肠道纵向蠕动的方案。我们描述了转运测试、整装免疫染色和组织采集的步骤。然后，我们详细说明了图像处理和手动细胞计数的程序。该方案旨在在评估可能是实验条件之间蠕动差异基础的细胞和分子特征时，尽量减少试验间的变异性。有关此方案的使用和执行的完整详细信息，请参考弗里思等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f942/12022682/df4aaed15e1b/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f942/12022682/047c026673b6/fx1.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f942/12022682/c0871a5fc505/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f942/12022682/480ae3f35d0d/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f942/12022682/51396e941248/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f942/12022682/bf2e66118e64/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f942/12022682/3a441cfbeb2c/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f942/12022682/57f311cd26bc/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f942/12022682/df4aaed15e1b/gr8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f942/12022682/047c026673b6/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f942/12022682/33dcfa8edb8c/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f942/12022682/c0871a5fc505/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f942/12022682/480ae3f35d0d/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f942/12022682/51396e941248/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f942/12022682/bf2e66118e64/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f942/12022682/3a441cfbeb2c/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f942/12022682/57f311cd26bc/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f942/12022682/df4aaed15e1b/gr8.jpg

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