Abstract：
A fundamental understanding of the enteric nervous system in normal and diseased states is limited by the lack of standard measures of total enteric neuron number. The adult zebrafish is a useful model in this context as it is amenable to in toto imaging of the intestine. We leveraged this to develop a technique to image and quantify all enteric neurons within the adult zebrafish intestine and applied this method to assess the relationship between intestinal length and total enteric neuron number.
Dissected adult zebrafish intestines were immunostained in wholemount, optically cleared with refractive index-matched solution, and then imaged in tiles using light-sheet microscopy. Imaging software was used to stitch the tiles, and the full image underwent automated cell counting. Total enteric neuron number was assessed in relation to intestinal length using linear regression modeling.
Whole gut imaging of the adult zebrafish intestine permits the visualization of endogenous and immunohistochemistry-derived fluorescence throughout the intestine. While enteric neuron distribution is heterogeneous between intestinal segments, total enteric neuron number positively correlates with intestinal length.
Imaging of all enteric neurons within the adult vertebrate intestine is possible in models such as the zebrafish. In this study, we apply this to demonstrate a positive correlation between enteric neuron number and intestinal length. Quantifying total enteric numbers will facilitate future studies of enteric neuropathies and ENS structure in animal models and potentially in biopsied tissue samples.
Dissected adult zebrafish intestines were immunostained in wholemount, optically cleared with refractive index-matched solution, and then imaged in tiles using light-sheet microscopy. Imaging software was used to stitch the tiles, and the full image underwent automated cell counting. Total enteric neuron number was assessed in relation to intestinal length using linear regression modeling.
Whole gut imaging of the adult zebrafish intestine permits the visualization of endogenous and immunohistochemistry-derived fluorescence throughout the intestine. While enteric neuron distribution is heterogeneous between intestinal segments, total enteric neuron number positively correlates with intestinal length.
Imaging of all enteric neurons within the adult vertebrate intestine is possible in models such as the zebrafish. In this study, we apply this to demonstrate a positive correlation between enteric neuron number and intestinal length. Quantifying total enteric numbers will facilitate future studies of enteric neuropathies and ENS structure in animal models and potentially in biopsied tissue samples.
摘要：
对正常和患病状态下的肠神经系统的基本理解受到缺乏总肠神经元数量的标准测量的限制。成年斑马鱼在这种情况下是有用的模型，因为它适合于肠的成像。我们利用这一点来开发一种技术来成像和量化成年斑马鱼肠道内的所有肠神经元，并应用这种方法来评估肠长度和总肠神经元数量之间的关系。
解剖的成年斑马鱼肠在整体上进行免疫染色，用折射率匹配的溶液进行光学清除，然后使用光片显微镜在瓷砖中成像。使用成像软件来缝合瓷砖，和完整的图像进行了自动细胞计数。使用线性回归模型评估肠神经元总数与肠长度的关系。
成年斑马鱼肠道的全肠成像允许观察整个肠道的内源性和免疫组织化学衍生的荧光。虽然肠神经元分布在肠段之间是异质的，肠神经元总数与肠长度呈正相关。
成年脊椎动物肠道内所有肠神经元的成像在斑马鱼等模型中是可能的。在这项研究中,我们将其应用于证明肠神经元数量和肠长度之间的正相关。定量总肠数量将有助于在动物模型中以及潜在地在活检组织样品中对肠神经病和ENS结构的未来研究。
解剖的成年斑马鱼肠在整体上进行免疫染色，用折射率匹配的溶液进行光学清除，然后使用光片显微镜在瓷砖中成像。使用成像软件来缝合瓷砖，和完整的图像进行了自动细胞计数。使用线性回归模型评估肠神经元总数与肠长度的关系。
成年斑马鱼肠道的全肠成像允许观察整个肠道的内源性和免疫组织化学衍生的荧光。虽然肠神经元分布在肠段之间是异质的，肠神经元总数与肠长度呈正相关。
成年脊椎动物肠道内所有肠神经元的成像在斑马鱼等模型中是可能的。在这项研究中,我们将其应用于证明肠神经元数量和肠长度之间的正相关。定量总肠数量将有助于在动物模型中以及潜在地在活检组织样品中对肠神经病和ENS结构的未来研究。
