Guardiola Ombretta, Andolfi Gennaro, Tirone Mario, Iavarone Francescopaolo, Brunelli Silvia, Minchiotti Gabriella
Stem Cell Fate Laboratory, Institute of Genetics and Biophysics "A. Buzzati Traverso", CNR;
Stem Cell Fate Laboratory, Institute of Genetics and Biophysics "A. Buzzati Traverso", CNR.
J Vis Exp. 2017 Jan 1(119):54515. doi: 10.3791/54515.
Skeletal muscle regeneration is a physiological process that occurs in adult skeletal muscles in response to injury or disease. Acute injury-induced skeletal muscle regeneration is a widely used, powerful model system to study the events involved in muscle regeneration as well as the mechanisms and different players. Indeed, a detailed knowledge of this process is essential for a better understanding of the pathological conditions that lead to skeletal muscle degeneration, and it aids in identifying new targeted therapeutic strategies. The present work describes a detailed and reproducible protocol to induce acute skeletal muscle regeneration in mice through a single intramuscular injection of cardiotoxin (CTX). CTX belongs to the family of snake venom toxins and causes myolysis of myofibers, which eventually triggers the regeneration events. The dynamics of skeletal muscle regeneration is evaluated by histological analysis of muscle sections. The protocol also illustrates the experimental procedures for dissecting, freezing, and cutting the Tibialis Anterior muscle, as well as the routine Hematoxylin & Eosin staining that is widely used for subsequent morphological and morphometric analysis.
骨骼肌再生是一种生理过程，发生在成年骨骼肌中以应对损伤或疾病。急性损伤诱导的骨骼肌再生是一个广泛使用的、强大的模型系统，用于研究肌肉再生所涉及的事件以及相关机制和不同参与者。事实上，深入了解这一过程对于更好地理解导致骨骼肌退化的病理状况至关重要，并且有助于确定新的靶向治疗策略。本研究描述了一种详细且可重复的方案，通过单次肌肉注射心脏毒素（CTX）诱导小鼠急性骨骼肌再生。CTX属于蛇毒毒素家族，可导致肌纤维溶解，最终触发再生事件。通过对肌肉切片进行组织学分析来评估骨骼肌再生的动态过程。该方案还阐述了解剖、冷冻和切割胫骨前肌的实验步骤，以及广泛用于后续形态学和形态计量分析的常规苏木精和伊红染色。
