title: "Late" macroendosomes and acidic endosomes in vertebrate motor nerve terminals.
abstract: Activity at the vertebrate nerve-muscle synapse creates large macroendosomes (MEs) via bulk membrane infolding. Visualized with the endocytic probe FM1-43, most (94%) of the ∼25 MEs/terminal created by brief (30-Hz, 18-second) stimulation dissipate rapidly (∼1 minute) into vesicles. Others, however, remain for hours. Here we study these "late" MEs by using 4D live imaging over a period of ∼1 hour after stimulation. We find that some (51/398 or 13%) disappear spontaneously via exocytosis, releasing their contents into the extracellular milieu. Others (at least 15/1,960 or 1%) fuse or closely associate with a second class of endosomes that take up acidophilic dyes (acidic endosomes [AEs]). AEs are plentiful (∼47/terminal) and exist independent of stimulation. Unlike MEs, which exhibit Brownian motion, AEs exhibit directed motion (average, 83 nm/sec) on microtubules within and among terminal boutons. AEs populate the axon as well, where movement is predominantly retrograde. They share biochemical and immunohistochemical markers (e.g., lysosomal-associated membrane protein [LAMP-1]) with lysosomes. Fusion/association of MEs with AEs suggests a sorting/degradation pathway in nerve terminals wherein the role of AEs is similar to that of lysosomes. Based on our data, we propose that MEs serve as sorting endosomes. Thus their contents, which include plasma membrane proteins, vesicle proteins, and extracellular levels of Ca(2+) , can be targeted either toward the reformation and budding of synaptic vesicles, toward secretion via exocytosis, or toward a degradation process that utilizes AEs either for lysis within the terminal or for transport toward the cell body.
authors: Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110, USA. wilk@wustl.edu
authors: Richard S
authors: RS
authors: Stewart
authors: Haibing
authors: H
authors: Teng
authors: Robert S
authors: RS
authors: Wilkinson
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D002402:Cathepsin D / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003964:Diaphragm / Q000502:physiology; D004558:Electric Stimulation; D011992:Endosomes / Q000378:metabolism; D005089:Exocytosis / Q000502:physiology; D006801:Humans; D066298:In Vitro Techniques; D051907:Lysosomal Membrane Proteins / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D018613:Microscopy, Confocal; D018482:Muscle, Skeletal / Q000502:physiology; D009469:Neuromuscular Junction / Q000166:cytology* / Q000502:physiology*; D017729:Presynaptic Terminals / Q000502:physiology*; D012911:Snakes / Q000033:anatomy & histology
chemical_list: D051907:Lysosomal Membrane Proteins; D008562:Membrane Glycoproteins; D002402:Cathepsin D; D002118:Calcium