Oudhoff Hendrik, Baumgartner Florian, Jaźwińska Anna
Department of Biology, University of Fribourg.
Department of Biology, University of Fribourg;
J Vis Exp. 2023 Jul 7(197). doi: 10.3791/65491.
Skeletal muscle undergoes renewal and restoration after minor injury through the activation of satellite-like stem cells. Severe injuries of the musculature often lead to fibrosis in humans. In comparison to mammals, zebrafish possess a higher innate capacity for organ regeneration, providing a powerful model for studying tissue restoration after extensive damage to the organ. Here, a cryoinjury model is described to induce profound damage to four myomeres of the caudal peduncle in adult zebrafish. A custom-made cryoprobe was designed to fit the body shape and reproducibly injure the lateral musculature from the skin to the midline. Importantly, the body integrity remained intact, and the fish continued their swimming activity. Changes to the skeletal muscle were assessed by histological staining and fluorescence staining of sarcomeric proteins on tissue sections. This method will open up new avenues of research aiming to understand how the degeneration of the skeletal muscle induces reparative responses and, thus, the reactivation of the myogenic program in adult zebrafish.
骨骼肌在受到轻微损伤后，通过激活卫星样干细胞进行更新和修复。在人类中，肌肉组织的严重损伤往往会导致纤维化。与哺乳动物相比，斑马鱼具有更高的器官再生先天能力，为研究器官受到广泛损伤后的组织修复提供了一个有力的模型。在此，描述了一种冷冻损伤模型，用于诱导成年斑马鱼尾柄的四个肌节受到严重损伤。设计了一种定制的冷冻探头，以适应鱼的体型，并可重复地损伤从皮肤到中线的外侧肌肉组织。重要的是，鱼体完整性保持完好，鱼继续其游泳活动。通过对组织切片进行组织学染色和肌节蛋白荧光染色来评估骨骼肌的变化。该方法将开辟新的研究途径，旨在了解成年斑马鱼骨骼肌的退化如何诱导修复反应，从而重新激活肌源性程序。
