Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Cdk5-dependent regulation of glucose-stimulated insulin secretion

Abstract

Tight glycemic control in individuals with diabetes mellitus is essential to prevent or delay its complications1. Present treatments to reduce hyperglycemia mainly target the ATP-sensitive K+ (KATP) channel of pancreatic beta cells to increase insulin secretion. These current approaches are often associated with the side effect of hypoglycemia. Here we show that inhibition of the activity of cyclin-dependent kinase 5 (Cdk5) enhanced insulin secretion under conditions of stimulation by high glucose but not low glucose in MIN6 cells and pancreatic islets. The role of Cdk5 in regulation of insulin secretion was confirmed in pancreatic beta cells deficient in p35, an activator of Cdk5. p35-knockout mice also showed enhanced insulin secretion in response to a glucose challenge. Cdk5 kinase inhibition enhanced the inward whole-cell Ca2+ channel current and increased Ca2+ influx across the L-type voltage-dependent Ca2+ channel (L-VDCC) upon stimulation with high glucose in beta cells, but had no effect on Ca2+ influx without glucose stimulation. The inhibitory regulation by Cdk5 on the L-VDCC was attributed to the phosphorylation of loop II-III of the α1C subunit of L-VDCC at Ser783, which prevented the binding to SNARE proteins and subsequently resulted in a decrease of the activity of L-VDCC. These results suggest that Cdk5/p35 may be a drug target for the regulation of glucose-stimulated insulin secretion.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1: Cdk5 inhibition enhances glucose-stimulated insulin secretion in MIN6 cells.
Figure 2: Effect of Cdk5 inhibition on Ca2+ mobilization in beta cells.
Figure 3: Effect of phosphorylation of the LII-III domain on interactions with syntaxin and SNAP-25.
Figure 4: p35 deficiency promotes insulin secretion in isolated islets and in vivo.

Accession codes

Accessions

BINDPlus

References

  1. Moller, D.E. New drug targets for type 2 diabetes and the metabolic syndrome. Nature 414, 821–827 (2001).

    Article  CAS  Google Scholar 

  2. Inagaki, N. et al. Reconstitution of IKATP: an inward rectifier subunit plus the sulfonylurea receptor. Science 270, 1166–1170 (1995).

    Article  CAS  Google Scholar 

  3. Bratanova-Tochkova, T.K. et al. Triggering and augmentation mechanisms, granule pools, and biphasic insulin secretion. Diabetes 51, S83–S90 (2002).

    Article  CAS  Google Scholar 

  4. Tsai, L.H., Takahashi, T., Caviness, V.S. Jr . & Harlow, E. Activity and expression pattern of cyclin-dependent kinase 5 in the embryonic mouse nervous system. Development 119, 1029–1040 (1993).

    CAS  PubMed  Google Scholar 

  5. Lew, J. et al. A brain-specific activator of cyclin-dependent kinase 5. Nature 371, 423–426 (1994).

    Article  CAS  Google Scholar 

  6. Tsai, L.H., Delalle, I., Caviness, V.S. Jr ., Chae, T. & Harlow, E. p35 is a neural-specific regulatory subunit of cyclin-dependent kinase 5. Nature 371, 419–423 (1994).

    Article  CAS  Google Scholar 

  7. Tang, D. et al. An isoform of the neuronal cyclin-dependent kinase 5 (Cdk5) activator. J. Biol. Chem. 270, 26897–26903 (1995).

    Article  CAS  Google Scholar 

  8. Ubeda, M., Kemp, D.M. & Habener, J.F. Glucose-induced expression of the cyclin-dependent protein kinase 5 activator p35 involved in Alzheimer's disease regulates insulin gene transcription in pancreatic beta-cells. Endocrinology 145, 3023–3031 (2004).

    Article  CAS  Google Scholar 

  9. Lilja, L. et al. Cyclin-dependent kinase 5 associated with p39 promotes Munc18–1 phosphorylation and Ca2+-dependent exocytosis. J. Biol. Chem. 279, 29534–29541 (2004).

    Article  CAS  Google Scholar 

  10. Vesely, J. et al. Inhibition of cyclin-dependent kinases by purine analogues. Eur. J. Biochem. 224, 771–786 (1994).

    Article  CAS  Google Scholar 

  11. Ozaki, N. et al. cAMP-GEFII is a direct target of cAMP in regulated exocytosis. Nat. Cell Biol. 2, 805–811 (2000).

    Article  CAS  Google Scholar 

  12. Ohshima, T. et al. Synergistic contributions of cyclin-dependant kinase 5/p35 and Reelin/Dab1 to the positioning of cortical neurons in the developing mouse brain. Proc. Natl. Acad. Sci. USA 98, 2764–2769 (2001).

    Article  CAS  Google Scholar 

  13. Chae, T. et al. Mice lacking p35, a neuronal specific activator of Cdk5, display cortical lamination defects, seizures, and adult lethality. Neuron 18, 29–42 (1997).

    Article  CAS  Google Scholar 

  14. Tomizawa, K. et al. Cdk5/p35 regulates neurotransmitter release through phosphorylation and downregulation of P/Q-type voltage-dependent calcium channel activity. J. Neurosci. 22, 2590–2597 (2002).

    Article  CAS  Google Scholar 

  15. Schulla, V. et al. Impaired insulin secretion and glucose tolerance in β cell-selective CaV1.2 Ca2+ channel null mice. EMBO J. 22, 3844–3854 (2003).

    Article  CAS  Google Scholar 

  16. Wiser, O., Bennett, M.K. & Atlas, D. Functional interaction of syntaxin and SNAP-25 with voltage-sensitive L- and N-type Ca2+ channels. EMBO J. 15, 4100–4110 (1996).

    Article  CAS  Google Scholar 

  17. Yokoyama, C.T., Sheng, Z.H. & Catterall, W.A. Phosphorylation of the synaptic protein interaction site on N-type calcium channels inhibits interactions with SNARE proteins. J. Neurosci. 17, 6929–6938 (1997).

    Article  CAS  Google Scholar 

  18. Lilla, V. et al. Differential gene expression in well-regulated and dysregulated pancreatic β-cell (MIN6) sublines. Endocrinology 144, 1368–1379 (2003).

    Article  CAS  Google Scholar 

  19. Minami, K. et al. Insulin secretion and differential gene expression in glucose-responsive and -unresponsive MIN6 sublines. Am. J. Physiol. Endocrinol. Metab. 279, E773–781 (2000).

    Article  CAS  Google Scholar 

  20. Wollheim, C.B., Meda, P. & Halban, P.A. Isolation of pancreatic islets and primary culture of the intact microorgans or of dispersed islet cells. Methods Enzymol. 192, 188–223 (1990).

    Article  CAS  Google Scholar 

  21. Kawaki, J. et al. Unresponsiveness to glibenclamide during chronic treatment induced by reduction of ATP-sensitive K+ channel activity. Diabetes 48, 2001–2006 (1999).

    Article  CAS  Google Scholar 

  22. Tomizawa, K. et al. Cophosphorylation of amphiphysin I and dynamin I by Cdk5 regulates clathrin-mediated endocytosis of synaptic vesicles. J. Cell Biol. 163, 813–824 (2003).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank V. Lilla (Louis-Jeantet Research Laboratory, University Medical Center) for providing the MIN6 B1 cell line, H. Noguchi for the technical advice, and T. Ogawa and A. Kemori for technical assistance. This work was supported by a Grant-in-Aid for Scientific Research on Priority Areas “Membrane Traffic” from the Ministry of Education, Science, Sports and Culture of Japan.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Kazuhito Tomizawa.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Suppression of Cdk5 expression level by siRNA enhanced insulin secretion in MIN6 cells. (PDF 152 kb)

Supplementary Fig. 2

Reduced phosphorylation level of Ser783 of αlc in the brain and islets of p35-deficient mice. (PDF 184 kb)

Supplementary Fig. 3

Immunostaining of islets in wild-type and p35-knockout mice. (PDF 1086 kb)

Supplementary Fig. 4

Islet mass of wild-type and p35-deficient mice. (PDF 2126 kb)

Supplementary Methods (PDF 72 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Wei, FY., Nagashima, K., Ohshima, T. et al. Cdk5-dependent regulation of glucose-stimulated insulin secretion. Nat Med 11, 1104–1108 (2005). https://doi.org/10.1038/nm1299

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm1299

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing