TY - JOUR
T1 - Synaptojanin and Endophilin Mediate Neck Formation during Ultrafast Endocytosis
AU - Watanabe, Shigeki
AU - Mamer, Lauren Elizabeth
AU - Raychaudhuri, Sumana
AU - Luvsanjav, Delgermaa
AU - Eisen, Julia
AU - Trimbuch, Thorsten
AU - Söhl-Kielczynski, Berit
AU - Fenske, Pascal
AU - Milosevic, Ira
AU - Rosenmund, Christian
AU - Jorgensen, Erik M.
N1 - Funding Information:
We would like to thank Marta Orlando, Shu-Wen Chang, Marcial Camacho-Pérez, Tanja Rosenmund, Katja Pötschke, Bettina Brokowski, Rike Dannenberg, Sabine Lenz, and the Charité Viral Core Facility for technical assistance; Louis Kerr for providing access to electron microscopy at the Marine Biological Laboratory; and Pietro de Camilli and Timothy A. Ryan for reagents and lively discussions. We thank the Tzagournis Medical Research Endowment Fund for the Ohio State University College of Medicine Medical Student Research Scholarship (to L.E.M.). We thank EMBO ( ASTF 443-2012 ) and the Grass Foundation for providing a fellowship (to S.W.). The research was funded by the Johns Hopkins University start-up funds (to S.W.), the Johns Hopkins University discovery award (to S.W.), the University of Chicago – MBL Lillie award (to S.W., E.M.J., and C.R.), the National Science Foundation ( 1727271 to S.W.), the NIH ( NS034307 to E.M.J.), a European Research Council grant ( 249939 SYNVGLUT to C.R.), and German Research Council grants ( Neurocure EXC 257 to E.M.J. and C.R., SFB 665 and SFB 958 to C.R., and SFB889 and Emmy Noether to I.M.). S.W. is an Alfred P. Sloan fellow. E.M.J. is an Investigator of the Howard Hughes Medical Institute and an Alexander von Humboldt Scholar.
Funding Information:
We would like to thank Marta Orlando, Shu-Wen Chang, Marcial Camacho-Pérez, Tanja Rosenmund, Katja Pötschke, Bettina Brokowski, Rike Dannenberg, Sabine Lenz, and the Charité Viral Core Facility for technical assistance; Louis Kerr for providing access to electron microscopy at the Marine Biological Laboratory; and Pietro de Camilli and Timothy A. Ryan for reagents and lively discussions. We thank the Tzagournis Medical Research Endowment Fund for the Ohio State University College of Medicine Medical Student Research Scholarship (to L.E.M.). We thank EMBO (ASTF 443-2012) and the Grass Foundation for providing a fellowship (to S.W.). The research was funded by the Johns Hopkins University start-up funds (to S.W.), the Johns Hopkins University discovery award (to S.W.), the University of Chicago – MBL Lillie award (to S.W., E.M.J., and C.R.), the National Science Foundation (1727271 to S.W.), the NIH (NS034307 to E.M.J.), a European Research Council grant (249939 SYNVGLUT to C.R.), and German Research Council grants (Neurocure EXC 257 to E.M.J. and C.R., SFB 665 and SFB 958 to C.R., and SFB889 and Emmy Noether to I.M.). S.W. is an Alfred P. Sloan fellow. E.M.J. is an Investigator of the Howard Hughes Medical Institute and an Alexander von Humboldt Scholar.
Publisher Copyright:
© 2018 Elsevier Inc.
PY - 2018/6/27
Y1 - 2018/6/27
N2 - Ultrafast endocytosis generates vesicles from the plasma membrane as quickly as 50 ms in hippocampal neurons following synaptic vesicle fusion. The molecular mechanism underlying the rapid maturation of these endocytic pits is not known. Here we demonstrate that synaptojanin-1, and its partner endophilin-A, function in ultrafast endocytosis. In the absence of synaptojanin or endophilin, the membrane is rapidly invaginated, but pits do not become constricted at the base. The 5-phosphatase activity of synaptojanin is involved in formation of the neck, but 4-phosphatase is not required. Nevertheless, these pits are eventually cleaved into vesicles; within a 30-s interval, synaptic endosomes form and are resolved by clathrin-mediated budding. Then synaptojanin and endophilin function at a second step to aid with the removal of clathrin coats from the regenerated vesicles. These data together suggest that synaptojanin and endophilin can mediate membrane remodeling on a millisecond timescale during ultrafast endocytosis. Ultrafast endocytosis is a distinct form of synaptic vesicle recovery occurring within milliseconds of vesicle fusion. Using flash-and-freeze electron microscopy, Watanabe et al. demonstrate dual roles for synaptojanin and endophilin in membrane remodeling and clathrin uncoating on rapid timescales during ultrafast endocytosis.
AB - Ultrafast endocytosis generates vesicles from the plasma membrane as quickly as 50 ms in hippocampal neurons following synaptic vesicle fusion. The molecular mechanism underlying the rapid maturation of these endocytic pits is not known. Here we demonstrate that synaptojanin-1, and its partner endophilin-A, function in ultrafast endocytosis. In the absence of synaptojanin or endophilin, the membrane is rapidly invaginated, but pits do not become constricted at the base. The 5-phosphatase activity of synaptojanin is involved in formation of the neck, but 4-phosphatase is not required. Nevertheless, these pits are eventually cleaved into vesicles; within a 30-s interval, synaptic endosomes form and are resolved by clathrin-mediated budding. Then synaptojanin and endophilin function at a second step to aid with the removal of clathrin coats from the regenerated vesicles. These data together suggest that synaptojanin and endophilin can mediate membrane remodeling on a millisecond timescale during ultrafast endocytosis. Ultrafast endocytosis is a distinct form of synaptic vesicle recovery occurring within milliseconds of vesicle fusion. Using flash-and-freeze electron microscopy, Watanabe et al. demonstrate dual roles for synaptojanin and endophilin in membrane remodeling and clathrin uncoating on rapid timescales during ultrafast endocytosis.
KW - clathrin uncoating
KW - endophilin
KW - flash-and-freeze
KW - membrane remodeling
KW - synaptic endosomes
KW - synaptic vesicle endocytosis
KW - synaptic vesicle recycling
KW - synaptojanin
KW - ultrafast endocytosis
UR - http://www.scopus.com/inward/record.url?scp=85048792741&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85048792741&partnerID=8YFLogxK
U2 - 10.1016/j.neuron.2018.06.005
DO - 10.1016/j.neuron.2018.06.005
M3 - Article
C2 - 29953872
AN - SCOPUS:85048792741
SN - 0896-6273
VL - 98
SP - 1184-1197.e6
JO - Neuron
JF - Neuron
IS - 6
ER -