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DOC2 isoforms play dual roles in insulin secretion and insulin-stimulated glucose uptake

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Abstract

Aims/hypothesis

Glucose-stimulated insulin secretion (GSIS) and insulin-stimulated glucose uptake are processes that rely on regulated intracellular vesicle transport and vesicle fusion with the plasma membrane. DOC2A and DOC2B are calcium-sensitive proteins that were identified as key components of vesicle exocytosis in neurons. Our aim was to investigate the role of DOC2 isoforms in glucose homeostasis, insulin secretion and insulin action.

Methods

DOC2 expression was measured by RT-PCR and western blotting. Body weight, glucose tolerance, insulin action and GSIS were assessed in wild-type (WT), Doc2a −/− (Doc2aKO), Doc2b −/− (Doc2bKO) and Doc2a −/−/Doc2b −/− (Doc2a/Doc2bKO) mice in vivo. In vitro GSIS and glucose uptake were assessed in isolated tissues, and exocytotic proteins measured by western blotting. GLUT4 translocation was assessed by epifluorescence microscopy.

Results

Doc2b mRNA was detected in all tissues tested, whereas Doc2a was only detected in islets and the brain. Doc2aKO and Doc2bKO mice had minor glucose intolerance, while Doc2a/Doc2bKO mice showed pronounced glucose intolerance. GSIS was markedly impaired in Doc2a/Doc2bKO mice in vivo, and in isolated Doc2a/Doc2bKO islets in vitro. In contrast, Doc2bKO mice had only subtle defects in insulin secretion in vivo. Insulin action was impaired to a similar degree in both Doc2bKO and Doc2a/Doc2bKO mice. In vitro insulin-stimulated glucose transport and GLUT4 vesicle fusion were defective in adipocytes derived from Doc2bKO mice. Surprisingly, insulin action was not altered in muscle isolated from DOC2-null mice.

Conclusions/interpretation

Our study identifies a critical role for DOC2B in insulin-stimulated glucose uptake in adipocytes, and for the synergistic regulation of GSIS by DOC2A and DOC2B in beta cells.

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Abbreviations

DOC2:

Double C2 domain protein

Doc2aKO:

Doc2a −/−

Doc2bKO:

Doc2b −/−

DXA:

Dual-energy x-ray absorptiometry

GSIS:

Glucose-stimulated insulin secretion

ITT:

Insulin tolerance test

[Ca2+]i :

Intracellular calcium

MEFs:

Mouse embryonic fibroblasts

PM:

Plasma membrane

SNARE:

Soluble N-ethylmaleimide-sensitive factor attachment protein receptor

WT:

Wild-type

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Acknowledgements

We thank T. Südhof (Stanford University, CA, USA) for providing antibodies. We thank M. van de Bunt and A. Gloyn (University of Oxford, UK) for assistance with RNA-seq data analysis.

Funding

This work was supported by a National Health and Medical Research Council of Australia (NHMRC) program grant (DEJ) and project grant (JC), and by the Chinese Scholarship Council (JL). DEJ is an NHMRC Senior Principal Research Fellow.

Duality of interest

The authors declare they have no duality of interest associated with this manuscript.

Contribution statement

DEJ conceived the studies. JC wrote the manuscript, with critical input from all authors. DEJ, JC, JL, JGB and JS designed experiments and interpreted data. JC, JL, JGB, JS, CCM, PTW, HP and RC designed experiments, and acquired and analysed data. MV and AJAG contributed to the acquisition and interpretation of data. All authors approved the final manuscript. DEJ is the guarantor of this work.

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Authors and Affiliations

  1. Diabetes and Obesity Research Program, Garvan Institute of Medical Research, Darlinghurst, NSW, Australia

    Jia Li, James Cantley, James G. Burchfield, Christopher C. Meoli, Jacqueline Stöckli, P. Tess Whitworth, Himani Pant, Rima Chaudhuri & David E. James

  2. Department of Physiology, Fourth Military Medical University, Xi’an, Shaanxi Province, People’s Republic of China

    Jia Li

  3. Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, UK

    James Cantley

  4. St. Vincent’s Clinical School, Faculty of Medicine, University of New South Wales, Darlinghurst, NSW, Australia

    James Cantley & Jacqueline Stöckli

  5. Department of Functional Genomics and Department of Clinical Genetics, Center for Neurogenomics and Cognitive Research, Neuroscience Campus Amsterdam, VU University and VU Medical Center, Amsterdam, the Netherlands

    Alexander J. A. Groffen & Matthijs Verhage

  6. The Charles Perkins Centre, School of Molecular Biosciences, School of Medicine, University of Sydney, Sydney, NSW, 2006, Australia

    David E. James

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  1. Jia Li
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Correspondence to David E. James.

Additional information

Jia Li, James Cantley and James G. Burchfield contributed equally to this study.

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Li, J., Cantley, J., Burchfield, J.G. et al. DOC2 isoforms play dual roles in insulin secretion and insulin-stimulated glucose uptake. Diabetologia 57, 2173–2182 (2014). https://doi.org/10.1007/s00125-014-3312-y

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