title: Deciphering dynamics of clathrin-mediated endocytosis in a living organism.
abstract: Current understanding of clathrin-mediated endocytosis (CME) dynamics is based on detection and tracking of fluorescently tagged clathrin coat components within cultured cells. Because of technical limitations inherent to detection and tracking of single fluorescent particles, CME dynamics is not characterized in vivo, so the effects of mechanical cues generated during development of multicellular organisms on formation and dissolution of clathrin-coated structures (CCSs) have not been directly observed. Here, we use growth rates of fluorescence signals obtained from short CCS intensity trace fragments to assess CME dynamics. This methodology does not rely on determining the complete lifespan of individual endocytic assemblies. Therefore, it allows for real-time monitoring of spatiotemporal changes in CME dynamics and is less prone to errors associated with particle detection and tracking. We validate the applicability of this approach to in vivo systems by demonstrating the reduction of CME dynamics during dorsal closure of Drosophila melanogaster embryos.
authors.affiliation: Department of Physics, The Ohio State University, Columbus, OH 43210.
authors.forename: Joshua P
authors.identifier: 0000-0002-7990-6387
authors.initials: JP
authors.lastname: Ferguson
authors.affiliation: Department of Physics, The Ohio State University, Columbus, OH 43210.
authors.forename: Nathan M
authors.identifier: 0000-0002-6090-1148
authors.initials: NM
authors.lastname: Willy
authors.affiliation: Department of Physics, The Ohio State University, Columbus, OH 43210.
authors.forename: Spencer P
authors.initials: SP
authors.lastname: Heidotting
authors.affiliation: Department of Physics, The Ohio State University, Columbus, OH 43210.
authors.forename: Scott D
authors.initials: SD
authors.lastname: Huber
authors.affiliation: Biophysics Graduate Program, The Ohio State University, Columbus, OH 43210.
authors.forename: Matthew J
authors.identifier: 0000-0003-1092-4410
authors.initials: MJ
authors.lastname: Webber
authors.affiliation: Department of Physics, The Ohio State University, Columbus, OH 43210 Biophysics Graduate Program, The Ohio State University, Columbus, OH 43210 kural.1@osu.edu.
authors.forename: Comert
authors.identifier: 0000-0002-9065-6542
authors.initials: C
authors.lastname: Kural
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival; D058948:Cell Tracking; D002784:Cholesterol / Q000378:metabolism; D002966:Clathrin / Q000378:metabolism*; D022163:Clathrin-Coated Vesicles; D004331:Drosophila melanogaster / Q000166:cytology* / Q000196:embryology / Q000378:metabolism*; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000378:metabolism; D004705:Endocytosis; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000378:metabolism; D021621:Imaging, Three-Dimensional; D013997:Time Factors
chemical_list: D002966:Clathrin; D049452:Green Fluorescent Proteins; D002784:Cholesterol