Journal
NEURON
Volume 88, Issue 5, Pages -Publisher
CELL PRESS
DOI: 10.1016/j.neuron.2015.10.043
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Funding
- Medical Research Council [G1002117]
- Marie-Curie Initial Training Network grant [289581: NPlast]
- BBSRC PhD studentship
- Medical Research Council [G1002117] Funding Source: researchfish
- MRC [G1002117] Funding Source: UKRI
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The accurate formation of synaptic vesicles (SVs) and incorporation of their protein cargo during endocytosis is critical for the maintenance of neurotransmission. During intense neuronal activity, a transient and acute accumulation of SV cargo occurs at the plasma membrane. Activity-dependent bulk endocytosis (ADBE) is the dominant SV endocytosis mode under these conditions; however, it is currently unknown how ADBE mediates cargo retrieval. We examined the retrieval of different SV cargo molecules during intense stimulation using a series of genetically encoded pH-sensitive reporters in neuronal cultures. The retrieval of only one reporter, VAMP4-pHluorin, was perturbed by inhibiting ADBE. This selective recovery was confirmed by the enrichment of endogenous VAMP4 in purified bulk endosomes formed by ADBE. VAMP4 was also essential for ADBE, with a cytoplasmic di-leucine motif being critical for this role. Therefore, VAMP4 is the first identified ADBE cargo and is essential for this endocytosis mode to proceed.
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