Dap160/intersectin acts as a stabilizing scaffold required for synaptic development and vesicle endocytosis.
We describe the isolation of mutations in dynamin-associated protein 160 kDa (dap160), the Drosophila homolog of intersectin, a putative adaptor for proteins involved in endocytosis, cytoskeletal regulation, and signaling. We show that partial loss-of-function mutants display temperature-sensitive (ts) paralysis, whereas null mutants show ts defects in endocytosis. Loss-of-function mutants exhibit bouton overgrowth at larval neuromuscular junctions (NMJs), but evoked neurotransmission is normal. Mutant NMJs show a mild endocytic defect at 22 degrees C, which is strongly enhanced at 34 degrees C. The levels of dynamin, synaptojanin and endophilin are severely reduced in dap160 mutant NMJs, suggesting that Dap160 serves to stabilize an endocytic macromolecular complex. Electron microscopy reveals fewer vesicles, aberrant large vesicles, and an accumulation of endocytic intermediates at active and periactive zones in mutant terminals. Our data suggest that Dap160, like dynamin, is involved in synaptic vesicle retrieval at active and periactive zones.
Koh
Tong-Wey
TW
Program in Developmental Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA.
Verstreken
Patrik
P
Bellen
Hugo J
HJ
D033942:Adaptor Proteins, Vesicular Transport; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism / Q000502:physiology*; D004330:Drosophila / Q000254:growth & development; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D004705:Endocytosis / Q000502:physiology*; D007814:Larva; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D009043:Motor Activity / Q000235:genetics; D009048:Motor Skills / Q000502:physiology; D009154:Mutation; D009469:Neuromuscular Junction / Q000648:ultrastructure; D009479:Neuropeptides / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D013569:Synapses / Q000502:physiology* / Q000648:ultrastructure; D013572:Synaptic Vesicles / Q000502:physiology*; D013697:Thermosensing / Q000235:genetics; D033921:Vesicular Transport Proteins
D033942:Adaptor Proteins, Vesicular Transport; D002352:Carrier Proteins; C113692:Dap160 protein, Drosophila; D029721:Drosophila Proteins; D008565:Membrane Proteins; D009479:Neuropeptides; D033921:Vesicular Transport Proteins; C115919:intersectin 1