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Neurosci Res. 2019 Feb 18. pii: S0168-0102(18)30523-6. doi: 10.1016/j.neures.2019.02.004. [Epub ahead of print]

Confocal and multiphoton calcium imaging of the enteric nervous system in anesthetized mice.

Author information

1
Graduate School of Science and Engineering, Saitama University, Saitama, 338-8570, Japan; Brain and Body System Science Institute, Saitama University, Saitama, 338-8570, Japan.
2
Graduate School of Science and Engineering, Saitama University, Saitama, 338-8570, Japan; Brain and Body System Science Institute, Saitama University, Saitama, 338-8570, Japan; RIKEN Center for Brain Science, Wako, Saitama, 351-0198, Japan. Electronic address: masasato@mail.saitama-u.ac.jp.
3
Department of Anatomy, Division of Medicine, Faculty of Medical Sciences, University of Fukui, Fukui, 910-1193, Japan.
4
Graduate School of Science and Engineering, Saitama University, Saitama, 338-8570, Japan; Brain and Body System Science Institute, Saitama University, Saitama, 338-8570, Japan; Second Department of Pharmacology, School of Pharmaceutical Sciences, Kyushu University of Health and Welfare, Miyazaki, 882-8508, Japan.
5
Graduate School of Science and Engineering, Saitama University, Saitama, 338-8570, Japan; Brain and Body System Science Institute, Saitama University, Saitama, 338-8570, Japan; RIKEN Center for Brain Science, Wako, Saitama, 351-0198, Japan.
6
Laboratory of Chemical Pharmacology, Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo, 113-0033, Japan; Center for Information and Neural Networks, National Institute of Information and Communications Technology, Suita, Osaka, 565-0871, Japan.
7
Shionogi & Co., Ltd, Osaka, 561-0825, Japan.
8
Department of Orthopaedic Surgery, Nara Medical University, School of Medicine, Nara, 634-8521, Japan. Electronic address: mtakaki@naramed-u.ac.jp.
9
Graduate School of Science and Engineering, Saitama University, Saitama, 338-8570, Japan; Brain and Body System Science Institute, Saitama University, Saitama, 338-8570, Japan; RIKEN Center for Brain Science, Wako, Saitama, 351-0198, Japan; RIKEN Center for Advanced Intelligence Project, Tokyo, 103-0072, Japan. Electronic address: jnakai@mail.saitama-u.ac.jp.

Abstract

Most imaging studies of the enteric nervous system (ENS) that regulates the function of the gastrointestinal tract are so far performed using preparations isolated from animals, thus hindering the understanding of the ENS function in vivo. Here we report a method for imaging the ENS cellular network activity in living mice using a new transgenic mouse line that co-expresses G-CaMP6 and mCherry in the ENS combined with the suction-mediated stabilization of intestinal movements. With confocal or two-photon imaging, our method can visualize spontaneous and pharmacologically-evoked ENS network activity in living animals at cellular and subcellular resolutions, demonstrating the potential usefulness for studies of the ENS function in health and disease.

KEYWORDS:

Confocal microscopy; Genetically-encoded calcium indicator; Gut; In vivo imaging; Serotonin; Transgenic mice; Two-photon microscopy

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