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Published on: September 3, 2014
Dap160/intersectin acts as a stabilizing scaffold required for synaptic development and vesicle endocytosis
Tong-Wey Koh1, Patrik Verstreken, Hugo J Bellen
1Program in Developmental Biology, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA.
Neuron
|July 21, 2004
Summary
Mutations in Dap160, a protein linked to endocytosis, cause temperature-sensitive paralysis and synaptic defects in Drosophila. Dap160 is crucial for stabilizing endocytic machinery and synaptic vesicle retrieval.
Area of Science:
- Neuroscience
- Cell Biology
- Genetics
Background:
- Dynamin-associated protein 160 kDa (Dap160) is the Drosophila homolog of intersectin.
- Intersectin is a putative adaptor protein involved in endocytosis, cytoskeletal regulation, and signaling.
Purpose of the Study:
- To isolate and characterize mutations in Dap160.
- To investigate the role of Dap160 in endocytosis and synaptic function in Drosophila.
Main Methods:
- Isolation of Dap160 loss-of-function mutants.
- Analysis of temperature-sensitive paralysis and endocytic defects.
- Assessment of neuromuscular junction (NMJ) morphology and neurotransmission.
- Biochemical analysis of endocytic protein levels.
- Electron microscopy of mutant NMJs.
Main Results:
- Partial loss-of-function Dap160 mutants exhibit temperature-sensitive paralysis.
- Null mutants display temperature-sensitive defects in endocytosis.
- Dap160 mutants show bouton overgrowth at larval NMJs with normal evoked neurotransmission.
- Endocytic defects are mild at 22°C and enhanced at 34°C.
- Levels of dynamin, synaptojanin, and endophilin are reduced in Dap160 mutants.
- Electron microscopy reveals fewer vesicles, aberrant large vesicles, and endocytic intermediates accumulation.
Conclusions:
- Dap160 is essential for stabilizing the endocytic macromolecular complex.
- Dap160 plays a critical role in synaptic vesicle retrieval at active and periactive zones.
- Dap160 functions similarly to dynamin in synaptic vesicle recycling.
Keywords:
Non-programmatic
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