Connectomics Research Center, National Tsing Hua University Hsinchu, Taiwan.
Front Neuroanat. 2013 Oct 22;7:36. doi: 10.3389/fnana.2013.00036. eCollection 2013.
Tongue receives extensive innervation to perform taste, sensory, and motor functions. Details of the tongue neuroanatomy and its plasticity in response to injury offer insights to investigate tongue neurophysiology and pathophysiology. However, due to the dispersed nature of the neural network, standard histology cannot provide a global view of the innervation. We prepared transparent mouse tongue by optical clearing to reveal the spatial features of the tongue innervation and its remodeling in injury. Immunostaining of neuronal markers, including PGP9.5 (pan-neuronal marker), calcitonin gene-related peptide (sensory nerves), tyrosine hydroxylase (sympathetic nerves), and vesicular acetylcholine transporter (cholinergic parasympathetic nerves and neuromuscular junctions), was combined with vessel painting and nuclear staining to label the tissue network and architecture. The tongue specimens were immersed in the optical-clearing solution to facilitate photon penetration for 3-dimensiontal (3-D) confocal microscopy. Taking advantage of the transparent tissue, we simultaneously revealed the tongue microstructure and innervation with subcellular-level resolution. 3-D projection of the papillary neurovascular complex and taste bud innervation was used to demonstrate the spatial features of tongue mucosa and the panoramic imaging approach. In the tongue injury induced by 4-nitroquinoline 1-oxide administration in the drinking water, we observed neural tissue remodeling in response to the changes of mucosal and muscular structures. Neural networks and the neuromuscular junctions were both found rearranged at the peri-lesional region, suggesting the nerve-lesion interactions in response to injury. Overall, this new tongue histological approach provides a useful tool for 3-D imaging of neural tissues to better characterize their roles with the mucosal and muscular components in health and disease.
舌头接受广泛的神经支配，以执行味觉、感觉和运动功能。舌头神经解剖结构及其对损伤的可塑性的详细信息为研究舌头神经生理学和病理生理学提供了线索。然而，由于神经网络的分散性质，标准组织学不能提供神经支配的全局视图。我们通过光学透明化来制备透明的老鼠舌头，以揭示舌头神经支配及其在损伤后的重塑的空间特征。神经元标志物的免疫染色，包括 PGP9.5（神经元标志物）、降钙素基因相关肽（感觉神经）、酪氨酸羟化酶（交感神经）和囊泡乙酰胆碱转运体（胆碱能副交感神经和神经肌肉接头），与血管染色和核染色相结合，用于标记组织网络和结构。将舌头标本浸泡在光学透明溶液中，以促进光子穿透进行三维（3-D）共聚焦显微镜检查。利用透明组织，我们同时以亚细胞分辨率揭示了舌头的微观结构和神经支配。乳头神经血管复合体和味蕾神经支配的 3-D 投影用于展示舌头黏膜的空间特征和全景成像方法。在饮用水中给予 4-硝基喹啉 1-氧化物诱导的舌头损伤中，我们观察到神经组织对黏膜和肌肉结构变化的重塑。在损伤周边区域发现神经网络和神经肌肉接头都发生了重新排列，表明神经-损伤相互作用对损伤的反应。总的来说，这种新的舌头组织学方法为神经组织的 3-D 成像提供了一个有用的工具，以更好地描述它们在健康和疾病状态下与黏膜和肌肉成分的关系。
