In vivo imaging of enteric neurogenesis in the deep tissue of mouse small intestine.
In vivo imaging of enteric neurogenesis in the deep tissue of mouse small intestine.
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DOI：
10.1371/journal.pone.0054814
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Takaki M
中科院分区：
文献类型：
--
作者：
Goto K;Kato G;Kawahara I;Luo Y;Obata K;Misawa H;Ishikawa T;Kuniyasu H;Nabekura J;Takaki M
使用成像技术作为研究细胞生理学的工具的挑战之一是无法解析不位于制剂表面附近的结构。非线性光学显微镜，特别是双光子激发荧光显微镜(2 PM)克服了这一限制，在体外和体内制剂中提供了更深的光学穿透性(数百微米)。我们在肠道中使用这种方法，首次实现了H-line：Thy1启动子GFP小鼠小肠粘膜、粘膜下层、粘膜下层和肌间神经丛以及环行肌和纵行肌中肠神经元和神经纤维的活体成像。此外，我们还获得了肠切除和吻合术后新生的肠神经元的清晰三维成像。口服5-HT4受体激动剂柠檬酸莫沙必利(MOS)可促进神经发生。用药1周的小鼠新生神经元数为421±89个/864,900µm2(n = 5)，明显多于用药加拮抗剂组(113±76个/864,900µm2)和空白对照组(100±34个/864,900µm2)(n = 4)。大多数神经元位于表面100微米以内。上述结果证实，MOS激活肠神经5-HT4受体可促进新肠神经元的形成。我们的结论是，在活体内，2 PM成像使对活体小鼠整个肠道进行高分辨率深度成像成为可能，并揭示了5-HT4受体激活促进的新肠神经元的形成。
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-HT4-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 µm2 (n = 5), which was significantly greater than that observed in preparations treated with MOS plus an antagonist (113±76 per 864,900 µm2) or in 4 week vehicle controls (100±34 per 864,900 µm2) (n = 4 both). Most neurons were located within 100 µm of the surface. These results confirm that activation of enteric neural 5-HT4-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-HT4-receptor activation.
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影响因子：
15.9
作者：
Laranjeira, Catia;Sandgren, Katarina;Pachnis, Vassilis
通讯作者：
Pachnis, Vassilis
DOI：
10.1152/ajpgi.00497.2001
发表时间：
2002-07-01
影响因子：
4.5
作者：
Yamanouchi, M;Shimatani, H;Takaki, M
通讯作者：
Takaki, M
DOI：
10.1523/jneurosci.1145-09.2009
发表时间：
2009-08-05
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Liu MT;Kuan YH;Wang J;Hen R;Gershon MD
通讯作者：
Gershon MD
影响因子：
64.5
作者：
Zhou, Ruyun;Niwa, Shinsuke;Hirokawa, Nobutaka
通讯作者：
Hirokawa, Nobutaka
影响因子：
16.2
作者：
Feng, GP;Mellor, RH;Sanes, JR
通讯作者：
Sanes, JR