Merck

Paraffin-embedded and frozen sections of Drosophila adult muscles.

Journal of visualized experiments : JoVE (2011-01-06)
Mariya M Kucherenko, April K Marrone, Valentyna M Rishko, Andriy S Yatsenko, Annekatrin Klepzig, Halyna R Shcherbata
ABSTRACT

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.

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Chloroform, anhydrous, ≥99%, contains 0.5-1.0% ethanol as stabilizer