In vivo imaging of enteric neurogenesis in the deep tissue of mouse small intestine.

One of the challenges of using imaging techniques as a tool to study cellular physiology has been the inability to resolve structures that are not located near the surface of the preparation. Nonlinear optical microscopy, in particular two photon-excited fluorescence microscopy (2PM), has overcome t...

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Autores principales:	Kei Goto, Go Kato, Isao Kawahara, Yi Luo, Koji Obata, Hiromi Misawa, Tatsuya Ishikawa, Hiroki Kuniyasu, Junich Nabekura, Miyako Takaki
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/aeae729ec1ae45ea9b23809fa808ab6c
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spelling	oai:doaj.org-article:aeae729ec1ae45ea9b23809fa808ab6c2021-11-18T07:59:14ZIn vivo imaging of enteric neurogenesis in the deep tissue of mouse small intestine.1932-620310.1371/journal.pone.0054814https://doaj.org/article/aeae729ec1ae45ea9b23809fa808ab6c2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23382976/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203One of the challenges of using imaging techniques as a tool to study cellular physiology has been the inability to resolve structures that are not located near the surface of the preparation. Nonlinear optical microscopy, in particular two photon-excited fluorescence microscopy (2PM), has overcome this limitation, providing deeper optical penetration (several hundred µm) in ex vivo and in vivo preparations. We have used this approach in the gut to achieve the first in vivo imaging of enteric neurons and nerve fibers in the mucosa, submucosa, submucosal and myenteric plexuses, and circular and longitudinal muscles of the small intestine in H-line: Thy1 promoter GFP mice. Moreover, we obtained clear three-dimensional imaging of enteric neurons that were newly generated after gut transection and reanastomosis. Neurogenesis was promoted by oral application of the 5-HT(4)-receptor agonist, mosapride citrate (MOS). The number of newly generated neurons observed in mice treated with MOS for one week was 421±89 per 864,900 µm(2) (n = 5), which was significantly greater than that observed in preparations treated with MOS plus an antagonist (113±76 per 864,900 µm(2)) or in 4 week vehicle controls (100±34 per 864,900 µm(2)) (n = 4 both). Most neurons were located within 100 µm of the surface. These results confirm that activation of enteric neural 5-HT(4)-receptor by MOS promotes formation of new enteric neurons. We conclude that in vivo 2PM imaging made it possible to perform high-resolution deep imaging of the living mouse whole gut and reveal formation of new enteric neurons promoted by 5-HT(4)-receptor activation.Kei GotoGo KatoIsao KawaharaYi LuoKoji ObataHiromi MisawaTatsuya IshikawaHiroki KuniyasuJunich NabekuraMiyako TakakiPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 1, p e54814 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Kei Goto Go Kato Isao Kawahara Yi Luo Koji Obata Hiromi Misawa Tatsuya Ishikawa Hiroki Kuniyasu Junich Nabekura Miyako Takaki In vivo imaging of enteric neurogenesis in the deep tissue of mouse small intestine.
description	One of the challenges of using imaging techniques as a tool to study cellular physiology has been the inability to resolve structures that are not located near the surface of the preparation. Nonlinear optical microscopy, in particular two photon-excited fluorescence microscopy (2PM), has overcome this limitation, providing deeper optical penetration (several hundred µm) in ex vivo and in vivo preparations. We have used this approach in the gut to achieve the first in vivo imaging of enteric neurons and nerve fibers in the mucosa, submucosa, submucosal and myenteric plexuses, and circular and longitudinal muscles of the small intestine in H-line: Thy1 promoter GFP mice. Moreover, we obtained clear three-dimensional imaging of enteric neurons that were newly generated after gut transection and reanastomosis. Neurogenesis was promoted by oral application of the 5-HT(4)-receptor agonist, mosapride citrate (MOS). The number of newly generated neurons observed in mice treated with MOS for one week was 421±89 per 864,900 µm(2) (n = 5), which was significantly greater than that observed in preparations treated with MOS plus an antagonist (113±76 per 864,900 µm(2)) or in 4 week vehicle controls (100±34 per 864,900 µm(2)) (n = 4 both). Most neurons were located within 100 µm of the surface. These results confirm that activation of enteric neural 5-HT(4)-receptor by MOS promotes formation of new enteric neurons. We conclude that in vivo 2PM imaging made it possible to perform high-resolution deep imaging of the living mouse whole gut and reveal formation of new enteric neurons promoted by 5-HT(4)-receptor activation.
format	article
author	Kei Goto Go Kato Isao Kawahara Yi Luo Koji Obata Hiromi Misawa Tatsuya Ishikawa Hiroki Kuniyasu Junich Nabekura Miyako Takaki
author_facet	Kei Goto Go Kato Isao Kawahara Yi Luo Koji Obata Hiromi Misawa Tatsuya Ishikawa Hiroki Kuniyasu Junich Nabekura Miyako Takaki
author_sort	Kei Goto
title	In vivo imaging of enteric neurogenesis in the deep tissue of mouse small intestine.
title_short	In vivo imaging of enteric neurogenesis in the deep tissue of mouse small intestine.
title_full	In vivo imaging of enteric neurogenesis in the deep tissue of mouse small intestine.
title_fullStr	In vivo imaging of enteric neurogenesis in the deep tissue of mouse small intestine.
title_full_unstemmed	In vivo imaging of enteric neurogenesis in the deep tissue of mouse small intestine.
title_sort	in vivo imaging of enteric neurogenesis in the deep tissue of mouse small intestine.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/aeae729ec1ae45ea9b23809fa808ab6c
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In vivo imaging of enteric neurogenesis in the deep tissue of mouse small intestine.

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