在口腔中,舌头是支持吞咽食物和液体的重要肌肉器官。它负责味觉,基于它包含的许多不同的味蕾。舌病领域的研究需要合适的临床前模型。因此,健康的兔舌可以作为病理情况的基线和参考。有了这个考虑,我们涵盖了健康兔舌的固定和组织学染色以及免疫组织化学标记。在本技术说明中,讨论了固定剂的初始选择,比较福尔马林固定和随后的石蜡包埋与冷冻保存。此外,我们通过几个例子描述了福尔马林固定的抗原回收步骤的效果。最后,我们提供ECM标记胶原蛋白I,胶原蛋白III,纤连蛋白,α-SMA和弹性蛋白染色以及ki67的增殖状态和PAR-2蛋白表达作为舌切片炎症状态和伤害感受的标志物,主要来自舌体。从技术上讲,我们发现石蜡切片对胶原蛋白I的优越性,胶原蛋白III,纤连蛋白,KI67和α-SMA标记,用于选定的检测系统。至于ECM组件,固有层富含胶原蛋白和纤连蛋白,而舌头的肌肉体仅显示肌纤维之间的胶原蛋白和纤连蛋白阳性区域。此外,α-SMA在动脉和静脉壁中明显表达。另一方面,炎性标志物PAR-2在唾液腺中显著表达,并在一定程度上在血管壁中表达。在舌的排泄管中发现了特定的PAR-2表达。本技术说明旨在提供基线图像,可用于比较患病兔舌的病理状态以及种间比较,如老鼠或老鼠的舌头。最后,它可以用于与人类的情况进行比较。
In the oral cavity the tongue is an important muscular organ that supports the swallowing of food and liquids. It is responsible for the sense of taste, based on the many different taste buds it contains. Research in the field of tongue diseases demands for suitable preclinical models. The healthy rabbit tongue may therefore serve as baseline and reference for the pathological situation. With this consideration, we covered the fixation and histological stainings as well as the immunohistochemical labelling of the healthy rabbit tongue. In this technical note, initial choice of the fixative is discussed, with a comparison of formalin fixation and subsequent paraffin embedding versus cryopreservation. Moreover, we delineate the effect of an antigen retrieval step for formalin fixation by several examples. Finally, we provide ECM markers collagen I, collagen III, fibronectin, α-SMA and elastin staining as well as ki67 for proliferative status and PAR-2 protein expression as a marker for inflammatory status and nociception in tongue sections, mainly from the tongue body. Technically, we found superiority of paraffin sections for collagen I, collagen III, fibronectin, ki67 and α-SMA labelling, for selected detections systems. As for ECM components, the lamina propria was very rich in collagen and fibronectin, while the muscular body of the tongue showed only collagen and fibronectin positive areas between the muscle fibers. Moreover, α-SMA was clearly expressed in the walls of arteries and veins. The inflammatory marker PAR-2 on the other hand was prominently expressed in the salivary glands and to some extent in the walls of the vessels. Particular PAR-2 expression was found in the excretory ducts of the tongue. This technical note has the aim to provide baseline images that can be used to compare the pathological state of the diseased rabbit tongue as well as for inter-species comparison, such as mouse or rat tongue. Finally, it can be used for the comparison with the human situation.