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Black HL, Livingstone R, Mastick CC, et al. Knockout of syntaxin-4 in 3T3-L1 adipocytes reveals new insight into GLUT4 trafficking and adiponectin secretion. J Cell Sci. 2022;135(1)doi: 10.1242/jcs.258375.
Black, H. L., Livingstone, R., Mastick, C. C., Al Tobi, M., Taylor, H., Geiser, A., Stirrat, L., Kioumourtzoglou, D., Petrie, J. R., Boyle, J. G., Bryant, N. J., & Gould, G. W. (2022). Knockout of syntaxin-4 in 3T3-L1 adipocytes reveals new insight into GLUT4 trafficking and adiponectin secretion. Journal of cell science, 135(1), . https://doi.org/10.1242/jcs.258375
Black, Hannah L, et al. "Knockout of syntaxin-4 in 3T3-L1 adipocytes reveals new insight into GLUT4 trafficking and adiponectin secretion." Journal of cell science vol. 135,1 (2022). doi: https://doi.org/10.1242/jcs.258375
Black HL, Livingstone R, Mastick CC, Al Tobi M, Taylor H, Geiser A, Stirrat L, Kioumourtzoglou D, Petrie JR, Boyle JG, Bryant NJ, Gould GW. Knockout of syntaxin-4 in 3T3-L1 adipocytes reveals new insight into GLUT4 trafficking and adiponectin secretion. J Cell Sci. 2022 Dec 01;135(1). doi: 10.1242/jcs.258375. Epub 2022 Jan 10. PMID: 34859814.
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J Cell Sci. 2022 Dec 01;135(1). doi: 10.1242/jcs.258375. Epub 2022 Jan 10.

Knockout of syntaxin-4 in 3T3-L1 adipocytes reveals new insight into GLUT4 trafficking and adiponectin secretion.

Journal of cell science

Hannah L Black, Rachel Livingstone, Cynthia C Mastick, Mohammed Al Tobi, Holly Taylor, Angéline Geiser, Laura Stirrat, Dimitrios Kioumourtzoglou, John R Petrie, James G Boyle, Nia J Bryant, Gwyn W Gould

Affiliations

  1. Department of Biology and York Biomedical Research Institute, University of York. Heslington, York YO10 5DD, UK.
  2. Henry Wellcome Laboratory for Cell Biology, Institute for Molecular, Cellular and Systems Biology, College of Medical Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK.
  3. Department of Biology, University of Nevada Reno, 1664 N. Virginia Street, Reno, NV 89557, USA.
  4. Strathclyde Institute for Pharmacy and Biomedical Sciences, 161 Cathedral Street, University of Strathclyde, Glasgow G4 0RE, UK.
  5. Institute of Cardiovascular and Medical Sciences, University of Glasgow, Glasgow G12 8QQ. UK.
  6. School of Medicine, Dentistry and Nursing, University of Glasgow, Glasgow G12 8QQ, UK.

PMID: 34859814 DOI: 10.1242/jcs.258375

Abstract

Adipocytes are key to metabolic regulation, exhibiting insulin-stimulated glucose transport that is underpinned by the insulin-stimulated delivery of glucose transporter type 4 (SLC2A4, also known and hereafter referred to as GLUT4)-containing vesicles to the plasma membrane where they dock and fuse, and increase cell surface GLUT4 levels. Adipocytokines, such as adiponectin, are secreted via a similar mechanism. We used genome editing to knock out syntaxin-4, a protein reported to mediate fusion between GLUT4-containing vesicles and the plasma membrane in 3T3-L1 adipocytes. Syntaxin-4 knockout reduced insulin-stimulated glucose transport and adiponectin secretion by ∼50% and reduced GLUT4 levels. Ectopic expression of haemagglutinin (HA)-tagged GLUT4 conjugated to GFP showed that syntaxin-4-knockout cells retain significant GLUT4 translocation capacity, demonstrating that syntaxin-4 is dispensable for insulin-stimulated GLUT4 translocation. Analysis of recycling kinetics revealed only a modest reduction in the exocytic rate of GLUT4 in knockout cells, and little effect on endocytosis. These analyses demonstrate that syntaxin-4 is not always rate limiting for GLUT4 delivery to the cell surface. In sum, we show that syntaxin-4 knockout results in reduced insulin-stimulated glucose transport, depletion of cellular GLUT4 levels and inhibition of adiponectin secretion but has only modest effects on the translocation capacity of the cells. This article has an associated First Person interview with Hannah L. Black and Rachel Livingstone, joint first authors of the paper.

© 2022. Published by The Company of Biologists Ltd.

Keywords: GLUT4; Membrane trafficking; SNARE; Syntaxin

Conflict of interest statement

Competing interests The authors declare no competing or financial interests.

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