Immunofluorescence Characterization of Muscle and Hypodermal Antigens in Embryos of the Nematode Caenorhabditus Elegans
Curry, Alice M.
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Caenorhabditus elegans is a small, free-living nematode whose development has been extensively studied. Zygotes follow rigidly-determined cleavage patterns to become embryos of 558 cells. Embryos undergo extensive changes in cellular position and morphology , resulting in four distinct morphological stages: lima bean, comma, tadpole, and pretzel. These gross morphological changes reflect the specific structural changes which are taking place. Development of muscle cells during embryogenesis has previously been studied, using antibodies against a major muscle protein, myosin, in immunofluorescence microscopy. Patterns of anti-myosin fluorescence are distinctive for each stage, and can be used to determine both the locations of muscle cells and the developmental stage of embryos. The present study used anti-myosin in conjunction with a series of monoclonal antibodies (mAb's) against various muscle and hypodermal antigens. An antibody against hypodermal desmosomes outlined individual hypodermal cells. Using this mAb with anti-myosin as a reference, the fusion of several uninucleate hypodermal cells into a large syncytium was followed. Another mAb labeled intermediate filaments in those regions of the hypodermis which lie directly over the bodywall muscles. Antibodies to an antigen in the basment membrane which connects the hypodermis to the bodywall muscles showed that this antigen was always extracellular, and appeared to be a product of the muscle cells. To confirm this, temperature-sensitive mutants which arrest cleavage at less than 100 cells were stained with antibodies to muscle, hypodermal, and basement membrane antigens. These mutants are known to express tissue- specific proteins, even after arrest. Comparison of staining patterns revealed that the basement membrane is a muscle cell product.