VAMP4 Is an Essential Cargo Molecule for Activity-Dependent Bulk Endocytosis

Neuron. 2015 Dec 2;88(5):973-984. doi: 10.1016/j.neuron.2015.10.043. Epub 2015 Nov 19.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Action Potentials / genetics
  • Animals
  • Animals, Newborn
  • Cells, Cultured
  • Cerebellum / cytology
  • Embryo, Mammalian
  • Endocytosis / genetics
  • Endocytosis / physiology*
  • Endosomes / metabolism
  • Endosomes / ultrastructure
  • Female
  • Heterocyclic Compounds, 3-Ring / metabolism
  • Hippocampus / cytology
  • Male
  • Mice
  • Neurons / physiology*
  • Neurons / ultrastructure
  • R-SNARE Proteins / genetics
  • R-SNARE Proteins / metabolism*
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Rhodamines
  • Synaptic Vesicles / metabolism
  • Synaptic Vesicles / ultrastructure
  • Vesicular Glutamate Transport Proteins / metabolism


  • Heterocyclic Compounds, 3-Ring
  • R-SNARE Proteins
  • RNA, Small Interfering
  • Rhodamines
  • VAMP4 protein, rat
  • Vesicular Glutamate Transport Proteins
  • tetramethylrosamine