title: Inactivation of clathrin heavy chain inhibits synaptic recycling but allows bulk membrane uptake.
abstract: Synaptic vesicle reformation depends on clathrin, an abundant protein that polymerizes around newly forming vesicles. However, how clathrin is involved in synaptic recycling in vivo remains unresolved. We test clathrin function during synaptic endocytosis using clathrin heavy chain (chc) mutants combined with chc photoinactivation to circumvent early embryonic lethality associated with chc mutations in multicellular organisms. Acute inactivation of chc at stimulated synapses leads to substantial membrane internalization visualized by live dye uptake and electron microscopy. However, chc-inactivated membrane cannot recycle and participate in vesicle release, resulting in a dramatic defect in neurotransmission maintenance during intense synaptic activity. Furthermore, inactivation of chc in the context of other endocytic mutations results in membrane uptake. Our data not only indicate that chc is critical for synaptic vesicle recycling but they also show that in the absence of the protein, bulk retrieval mediates massive synaptic membrane internalization.
authors.affiliation: Department of Molecular and Developmental Genetics, VIB Flemish Institute for Biotechnology, 3000 Leuven, Belgium.
authors.forename: Jaroslaw
authors.initials: J
authors.lastname: Kasprowicz
authors.forename: Sabine
authors.initials: S
authors.lastname: Kuenen
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Miskiewicz
authors.forename: Ron L P
authors.initials: RL
authors.lastname: Habets
authors.forename: Liesbet
authors.initials: L
authors.lastname: Smitz
authors.forename: Patrik
authors.initials: P
authors.lastname: Verstreken