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An Approach to Monitor Exocytosis in White Adipocytes

  • Ali M. Komai
  • Man Mohan Shrestha
  • Saliha Musovic
  • Charlotta S. OlofssonEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2233)

Abstract

Exocytosis, the fusion of vesicles with the plasma membrane, can be measured with the patch-clamp technique as increases in membrane capacitance. Here we provide detailed information on how to monitor white adipocyte exocytosis using this method. We describe how to isolate the stromal vascular fraction of cells (SVF) within adipose tissue and how to differentiate SVF and cultured 3T3-L1 cells into adipocytes suitable for patch-clamp studies. We also give detailed protocols of how to record and analyze exocytosis in the differentiated cells.

Key words

Exocytosis White adipocyte Patch-clamp Membrane capacitance Whole-cell configuration 3T3-L1 adipocytes Stromal vascular cell fraction Cell isolation Adipocyte differentiation 

Notes

Acknowledgments

This work was supported by the Swedish Medical Research Council (grant 2013-7107). We thank Mr. Habz Nafar for the illustration in Fig. 7a.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2021

Authors and Affiliations

  • Ali M. Komai
    • 1
  • Man Mohan Shrestha
    • 1
  • Saliha Musovic
    • 1
  • Charlotta S. Olofsson
    • 1
    Email author
  1. 1.Department of Physiology/Metabolic Physiology, Institute of Neuroscience and Physiology, The Sahlgrenska AcademyUniversity of GothenburgGöteborgSweden

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