title: Detecting Autophagy in Caenorhabditis elegans Embryos Using Markers of P Granule Degradation.
abstract: Autophagy plays an active role during the early stages of embryogenesis in the nematode Caenorhabditis elegans. Although their exact function is unknown, P granules are ribonucleoprotein particles that play a role in germ cell specification. The localization of P granules is restricted to the germline precursor cells in wild-type embryos, as a result of their degradation in the somatic cell lineage. Autophagy is known to be required for the degradation of P granules, as mutations in various autophagy genes, including those encoding the adaptor SEPA-1 and the p62-like adaptor SQST-1, result in the accumulation of the P granule components PGL-1 and PGL-3 (termed PGL granules) in the somatic cells of C. elegans embryos. In this protocol, we present a methodology for using fusion reporters of SEPA-1, SQST-1, and PGL-1 that have aided in the identification of new genes for normal autophagy activity by screening for mutant animals that lack the degradation of these autophagy substrates.
authors.affiliation: Department of Biology, Queens College-CUNY, Flushing, New York 11367; The Graduate Center, The City University of New York, New York 10016.
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Palmisano
authors.affiliation: Department of Biology, Queens College-CUNY, Flushing, New York 11367; The Graduate Center, The City University of New York, New York 10016.
authors.forename: Alicia
authors.initials: A
authors.lastname: Meléndez
mesh_terms: D000818:Animals; D001343:Autophagy / Q000502:physiology*; D017173:Caenorhabditis elegans / Q000166:cytology* / Q000196:embryology*; D047108:Embryonic Development / Q000235:genetics / Q000502:physiology*; D005854:Germ Cells / Q000473:pathology*; D059748:Proteolysis; D016601:RNA-Binding Proteins / Q000378:metabolism*
chemical_list: D016601:RNA-Binding Proteins