Distribution across tissue layers of extrinsic nerves innervating the mouse colorectum - an in vitro anterograde tracing study.
Distribution across tissue layers of extrinsic nerves innervating the mouse colorectum - an in vitro anterograde tracing study.
复制标题
支配小鼠结肠直肠的外在神经的组织层分布——一项体外顺行追踪研究。
DOI：
10.1111/nmo.12419
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Gebhart,GeraldF
中科院分区：
文献类型：
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作者：
Brumovsky,PabloR;La,Jun-Ho;Gebhart,GeraldF
研究背景盆神经（pelvic nerve，PN）的顺行追踪和整体标本水平面显示是研究结直肠周围神经分布的基础。在这里，我们进行了类似的分析，但在低温切片的小鼠结肠直肠，允许更直接的可视化神经分布在所有组织layer.MethodsColorectal与附加PN从成年雄性BalbC小鼠解剖。通过细PN纤维的单纤维记录证实存在活跃传入。随后使用生物素酰胺（BTA）进行“散装”（所有纤维）顺行追踪。分别使用组织学和免疫组织化学技术对BTA阳性神经进行可视化，并评价与降钙素基因相关肽（CGRP）的共定位。Key ResultsBTA阳性神经分布于小鼠结肠直肠的各层，与肌间神经丛神经元接触，分布于肌肉层内，穿透器官更深处，与血管、粘膜下神经丛神经元接触，甚至穿透粘膜。几个追踪的轴突共定位CGRP，支持其传入性质。最后，顺行追踪的PN也暴露了丰富的BTA阳性神经的主要pelvic ganglion.Conclusions & InferencesWe目前的模式的神经支配的外源性轴突跨层的小鼠结肠直肠，包括不稳定的粘膜层。所提出的方法也可能是有用的，在分析周围神经的形态和生理之间的关联，针对不同层的结直肠。
BackgroundAnterogradein vitrotracing of the pelvic nerve (PN) and visualization in the horizontal plane in whole mount preparations has been fundamental in the analysis of distribution of peripheral nerves innervating the colorectum. Here, we performed a similar analysis, but in cryostat sections of the mouse colorectum, allowing for a more direct visualization of nerve distribution in all tissue layers.MethodsColorectum with attached PNs was dissected from adult male BalbC mice. Presence of active afferents was certified by single fiber recording of fine PN fibers. This was followed by ‘bulk’ (all fibers) anterograde tracing using biotinamide (BTA). Histo‐ and immunohistochemical techniques were used for visualization of BTA‐positive nerves, and evaluation of co‐localization with calcitonin gene‐related peptide (CGRP), respectively. Tissue was analyzed using confocal microscopy on transverse or longitudinal colorectum sections.Key ResultsAbundant BTA‐positive nerves spanning all layers of the mouse colorectum and contacting myenteric plexus neurons, distributing within the muscle layer, penetrating deeper into the organ and contacting blood vessels, submucosal plexus neurons or even penetrating the mucosa, were regularly detected. Several traced axons co‐localized CGRP, supporting their afferent nature. Finally, anterograde tracing of the PN also exposed abundant BTA‐positive nerves in the major pelvic ganglion.Conclusions & InferencesWe present the patterns of innervation of extrinsic axons across layers in the mouse colorectum, including the labile mucosal layer. The proposed approach could also be useful in the analysis of associations between morphology and physiology of peripheral nerves targeting the different layers of the colorectum.