Kucherenko Mariya M, Marrone April K, Rishko Valentyna M, Yatsenko Andriy S, Klepzig Annekatrin, Shcherbata Halyna R
Gene Expression and Signaling Research Group, Max Planck Institute for Biophysical Chemistry.
J Vis Exp. 2010 Dec 27(46):2438. doi: 10.3791/2438.
The molecular characterization of muscular dystrophies and myopathies in humans has revealed the complexity of muscle disease and genetic analysis of muscle specification, formation and function in model systems has provided valuable insight into muscle physiology. Therefore, identifying and characterizing molecular mechanisms that underlie muscle damage is critical. The structure of adult Drosophila multi-fiber muscles resemble vertebrate striated muscles (1) and the genetic tractability of Drosophila has made it a great system to analyze dystrophic muscle morphology and characterize the processes affecting muscular function in ageing adult flies (2). Here we present the histological technique for preparing paraffin-embedded and frozen sections of Drosophila thoracic muscles. These preparations allow for the tissue to be stained with classical histological stains and labeled with protein detecting dyes, and specifically cryosections are ideal for immunohistochemical detection of proteins in intact muscles. This allows for analysis of muscle tissue structure, identification of morphological defects, and detection of the expression pattern for muscle/neuron-specific proteins in Drosophila adult muscles. These techniques can also be slightly modified for sectioning of other body parts.
人类肌肉萎缩症和肌病的分子特征揭示了肌肉疾病的复杂性，而在模型系统中对肌肉特化、形成和功能进行遗传分析，为深入了解肌肉生理学提供了有价值的见解。因此，识别和表征肌肉损伤背后的分子机制至关重要。成年果蝇多纤维肌肉的结构类似于脊椎动物的横纹肌(1)，果蝇的遗传易处理性使其成为分析营养不良性肌肉形态以及表征影响成年果蝇肌肉功能过程的理想系统(2)。在此，我们展示了用于制备果蝇胸部肌肉石蜡包埋切片和冷冻切片的组织学技术。这些制备方法能使组织用经典组织学染色剂染色并用蛋白质检测染料标记，特别是冷冻切片非常适合对完整肌肉中的蛋白质进行免疫组织化学检测。这使得能够分析肌肉组织结构、识别形态缺陷以及检测果蝇成年肌肉中肌肉/神经元特异性蛋白质的表达模式。这些技术稍作修改后也可用于其他身体部位的切片。
