RRC ID 34878
Author Nikolova LS, Metzstein MM.
Title Intracellular lumen formation in Drosophila proceeds via a novel subcellular compartment.
Journal Development
Abstract Cellular tubes have diverse morphologies, including multicellular, unicellular and subcellular architectures. Subcellular tubes are found prominently within the vertebrate vasculature, the insect breathing system and the nematode excretory apparatus, but how such tubes form is poorly understood. To characterize the cellular mechanisms of subcellular tube formation, we have refined methods of high pressure freezing/freeze substitution to prepare Drosophila larvae for transmission electron microscopic (TEM) analysis. Using our methods, we have found that subcellular tube formation may proceed through a previously undescribed multimembrane intermediate composed of vesicles bound within a novel subcellular compartment. We have also developed correlative light/TEM procedures to identify labeled cells in TEM-fixed larval samples. Using this technique, we have found that Vacuolar ATPase (V-ATPase) and the V-ATPase regulator Rabconnectin-3 are required for subcellular tube formation, probably in a step resolving the intermediate compartment into a mature lumen. In general, our ultrastructural analysis methods could be useful for a wide range of cellular investigations in Drosophila larvae.
Volume 142(22)
Pages 3964-73
Published 2015-11-15
DOI 10.1242/dev.127902
PII dev.127902
PMID 26428009
MeSH Adaptor Proteins, Signal Transducing / metabolism* Animals Cell Compartmentation / physiology* Drosophila / growth & development* Drosophila Proteins / metabolism* Freeze Fracturing Intracellular Space / physiology* Larva / growth & development Larva / ultrastructure Microscopy, Electron, Transmission / methods* Vacuolar Proton-Translocating ATPases / metabolism*
IF 5.413
Times Cited 4