title: Functional characterization of retromer in GLUT4 storage vesicle formation and adipocyte differentiation.
abstract: Insulin-stimulated translocation of glucose transporter 4 (GLUT4) storage vesicles (GSVs), the specialized intracellular compartments within mature adipocytes, to the plasma membrane (PM) is a fundamental cellular process for maintaining glucose homeostasis. Using 2 independent adipocyte cell line models, human primary Simpson-Golabi-Behmel syndrome and mouse 3T3-L1 fibroblast cell lines, we demonstrate that the endosome-associated protein-sorting complex retromer colocalizes with GLUT4 on the GSVs by confocal microscopy in mature adipocytes. By use of both confocal microscopy and differential ultracentrifugation techniques, retromer is redistributed to the PM of mature adipocytes upon insulin stimulation. Furthermore, stable knockdown of the retromer subunit-vacuolar protein-sorting 35, or the retromer-associated protein sorting nexin 27, by lentivirus-delivered small hairpin RNA impaired the adipogenesis process when compared to nonsilence control. The knockdown of retromer decreased peroxisome proliferator activated receptor γ expression during differentiation, generating adipocytes with decreased levels of GSVs, lipid droplet accumulation, and insulin-stimulated glucose uptake. In conclusion, our study demonstrates a role for retromer in the GSV formation and adipogenesis.
authors.affiliation: *Institute for Molecular Biosciences, The University of Queensland, St. Lucia, Queensland, Australia; and Division of Pediatric Endocrinology and Diabetes, Department of Pediatrics and Adolescent Medicine, University of Ulm, Ulm, Germany.
authors.forename: Zhe
authors.initials: Z
authors.lastname: Yang
authors.affiliation: *Institute for Molecular Biosciences, The University of Queensland, St. Lucia, Queensland, Australia; and Division of Pediatric Endocrinology and Diabetes, Department of Pediatrics and Adolescent Medicine, University of Ulm, Ulm, Germany.
authors.forename: Lee Kian
authors.initials: LK
authors.lastname: Hong
authors.affiliation: *Institute for Molecular Biosciences, The University of Queensland, St. Lucia, Queensland, Australia; and Division of Pediatric Endocrinology and Diabetes, Department of Pediatrics and Adolescent Medicine, University of Ulm, Ulm, Germany.
authors.forename: Jordan
authors.initials: J
authors.lastname: Follett
authors.affiliation: *Institute for Molecular Biosciences, The University of Queensland, St. Lucia, Queensland, Australia; and Division of Pediatric Endocrinology and Diabetes, Department of Pediatrics and Adolescent Medicine, University of Ulm, Ulm, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Wabitsch
authors.affiliation: *Institute for Molecular Biosciences, The University of Queensland, St. Lucia, Queensland, Australia; and Division of Pediatric Endocrinology and Diabetes, Department of Pediatrics and Adolescent Medicine, University of Ulm, Ulm, Germany.
authors.forename: Nicholas A
authors.initials: NA
authors.lastname: Hamilton
authors.affiliation: *Institute for Molecular Biosciences, The University of Queensland, St. Lucia, Queensland, Australia; and Division of Pediatric Endocrinology and Diabetes, Department of Pediatrics and Adolescent Medicine, University of Ulm, Ulm, Germany.
authors.forename: Brett M
authors.initials: BM
authors.lastname: Collins
authors.affiliation: *Institute for Molecular Biosciences, The University of Queensland, St. Lucia, Queensland, Australia; and Division of Pediatric Endocrinology and Diabetes, Department of Pediatrics and Adolescent Medicine, University of Ulm, Ulm, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Bugarcic
authors.affiliation: *Institute for Molecular Biosciences, The University of Queensland, St. Lucia, Queensland, Australia; and Division of Pediatric Endocrinology and Diabetes, Department of Pediatrics and Adolescent Medicine, University of Ulm, Ulm, Germany r.teasdale@uq.edu.au.
authors.forename: Rohan D
authors.initials: RD
authors.lastname: Teasdale
mesh_terms: D041721:3T3-L1 Cells; D017667:Adipocytes / Q000378:metabolism* / Q000502:physiology*; D000818:Animals; D001145:Arrhythmias, Cardiac / Q000378:metabolism / Q000473:pathology; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism / Q000502:physiology; D011992:Endosomes / Q000378:metabolism / Q000502:physiology; D005347:Fibroblasts / Q000378:metabolism / Q000502:physiology; D055785:Gene Knockdown Techniques / Q000379:methods; D040181:Genetic Diseases, X-Linked / Q000378:metabolism / Q000473:pathology; D005877:Gigantism / Q000378:metabolism / Q000473:pathology; D005947:Glucose / Q000378:metabolism; D051275:Glucose Transporter Type 4 / Q000378:metabolism*; D006330:Heart Defects, Congenital / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007328:Insulin / Q000378:metabolism; D008607:Intellectual Disability / Q000378:metabolism / Q000473:pathology; D051379:Mice; D047495:PPAR gamma / Q000378:metabolism; D021381:Protein Transport / Q000502:physiology
chemical_list: D051275:Glucose Transporter Type 4; D007328:Insulin; D047495:PPAR gamma; D005947:Glucose
keywords: adipogenesis; endosome; plasma membrane; protein trafficking