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A pancreatic islet-specific microRNA regulates insulin secretion

Nature volume 432, pages 226–230 (2004)Cite this article

Abstract

MicroRNAs (miRNAs) constitute a growing class of non-coding RNAs that are thought to regulate gene expression by translational repression1. Several miRNAs in animals exhibit tissue-specific or developmental-stage-specific expression, indicating that they could play important roles in many biological processes2,3,4. To study the role of miRNAs in pancreatic endocrine cells we cloned and identified a novel, evolutionarily conserved and islet-specific miRNA (miR-375). Here we show that overexpression of miR-375 suppressed glucose-induced insulin secretion, and conversely, inhibition of endogenous miR-375 function enhanced insulin secretion. The mechanism by which secretion is modified by miR-375 is independent of changes in glucose metabolism or intracellular Ca2+-signalling but correlated with a direct effect on insulin exocytosis. Myotrophin (Mtpn) was predicted to be and validated as a target of miR-375. Inhibition of Mtpn by small interfering (si)RNA mimicked the effects of miR-375 on glucose-stimulated insulin secretion and exocytosis. Thus, miR-375 is a regulator of insulin secretion and may thereby constitute a novel pharmacological target for the treatment of diabetes.

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Figure 1: miR-375 is expressed in pancreatic β-cells and regulates insulin secretion.
Figure 2: Expression of miR-375 using recombinant adenovirus (Ad-375) leads to impaired glucose-, KCl- and tolbutamide-induced insulin secretion in MIN6 cells.
Figure 3: No effect of miR-375 on intracellular Ca2+ signalling in β-cells.
Figure 4: Identification of target genes of miR-375.

References

  1. Bartel, D. P. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116, 281–297 (2004)

    Article  CAS  Google Scholar 

  2. Lagos-Quintana, M., Rauhut, R., Lendeckel, W. & Tuschl, T. Identification of novel genes coding for small expressed RNAs. Science 294, 853–858 (2001)

    Article  ADS  CAS  Google Scholar 

  3. Lau, N. C., Lim, L. P., Weinstein, E. G. & Bartel, D. P. An abundant class of tiny RNAs with probable regulatory roles in Caenorhabditis elegans. Science 294, 858–862 (2001)

    Article  ADS  CAS  Google Scholar 

  4. Lee, R. C. & Ambros, V. An extensive class of small RNAs in Caenorhabditis elegans. Science 294, 862–864 (2001)

    Article  ADS  CAS  Google Scholar 

  5. Carrington, J. C. & Ambros, V. Role of microRNAs in plant and animal development. Science 301, 336–338 (2003)

    Article  ADS  CAS  Google Scholar 

  6. Wightman, B., Ha, I. & Ruvkun, G. Posttranscriptional regulation of the heterochronic gene lin-14 by lin-4 mediates temporal pattern formation in C. elegans. Cell 75, 855–862 (1993)

    Article  CAS  Google Scholar 

  7. Olsen, P. H. & Ambros, V. The lin-4 regulatory RNA controls developmental timing in Caenorhabditis elegans by blocking LIN-14 protein synthesis after the initiation of translation. Dev. Biol. 216, 671–680 (1999)

    Article  CAS  Google Scholar 

  8. Brennecke, J., Hipfner, D. R., Stark, A., Russell, R. B. & Cohen, S. M. bantam encodes a developmentally regulated microRNA that controls cell proliferation and regulates the proapoptotic gene hid in Drosophila. Cell 113, 25–36 (2003)

    Article  CAS  Google Scholar 

  9. Xu, P., Vernooy, S. Y., Guo, M. & Hay, B. A. The Drosophila microRNA Mir-14 suppresses cell death and is required for normal fat metabolism. Curr. Biol. 13, 790–795 (2003)

    Article  CAS  Google Scholar 

  10. Pfeffer, S., Lagos-Quintana, M. & Tuschl, T. in Current Protocols in Molecular Biology (eds Ausubel, F. M. B. R. et al.) Ch. 26.4.1–26.4.18 (Wiley Interscience, New York, 2003)

    Google Scholar 

  11. Ambros, V. et al. A uniform system for microRNA annotation. RNA 9, 277–279 (2003)

    Article  CAS  Google Scholar 

  12. Meister, G., Landthaler, M., Dorsett, Y. & Tuschl, T. Sequence-specific inhibition of microRNA- and siRNA-induced RNA silencing. RNA 10, 544–550 (2004)

    Article  CAS  Google Scholar 

  13. Hutvagner, G., Simard, M. J., Mello, C. C. & Zamore, P. D. Sequence-specific inhibition of small RNA function. PLoS Biol 2, E98 (2004)

    Article  Google Scholar 

  14. Barg, S., Galvanovskis, J., Gopel, S. O., Rorsman, P. & Eliasson, L. Tight coupling between electrical activity and exocytosis in mouse glucagon-secreting alpha-cells. Diabetes 49, 1500–1510 (2000)

    Article  CAS  Google Scholar 

  15. Gopel, S. O., Kanno, T., Barg, S. & Rorsman, P. Patch-clamp characterisation of omatostatin-secreting cells in intact mouse pancreatic islets. J. Physiol. 528, 497–507 (2000)

    Article  CAS  Google Scholar 

  16. Tsuboi, T., da Silva Xavier, G., Leclerc, I. & Rutter, G. A. 5′-AMP-activated protein kinase controls insulin-containing secretory vesicle dynamics. J. Biol. Chem. 278, 52042–52051 (2003)

    Article  CAS  Google Scholar 

  17. Rajewsky, N. & Socci, N. D. Computational identification of microRNA targets. Dev. Biol. 267, 529–535 (2004)

    Article  CAS  Google Scholar 

  18. Antonin, W., Riedel, D. & von Mollard, G. F. The SNARE Vti1a-β is localized to small synaptic vesicles and participates in a novel SNARE complex. J. Neurosci. 20, 5724–5732 (2000)

    Article  CAS  Google Scholar 

  19. Taoka, M. et al. V-1, a protein expressed transiently during murine cerebellar development, regulates actin polymerization via interaction with capping protein. J. Biol. Chem. 278, 5864–5870 (2003)

    Article  CAS  Google Scholar 

  20. Larsen, C. M. et al. Interleukin-1β-induced rat pancreatic islet nitric oxide synthesis requires both the p38 and extracellular signal-regulated kinase 1/2 mitogen-activated protein kinases. J. Biol. Chem. 273, 15294–15300 (1998)

    Article  CAS  Google Scholar 

  21. Zhao, C., Wilson, M. C., Schuit, F., Halestrap, A. P. & Rutter, G. A. Expression and distribution of lactate/monocarboxylate transporter isoforms in pancreatic islets and the exocrine pancreas. Diabetes 50, 361–366 (2001)

    Article  CAS  Google Scholar 

  22. Schreiber-Agus, N. et al. An amino-terminal domain of Mxi1 mediates anti-Myc oncogenic activity and interacts with a homolog of the yeast transcriptional repressor SIN3. Cell 80, 777–786 (1995)

    Article  CAS  Google Scholar 

  23. Yamakuni, T. et al. V-1, a catecholamine biosynthesis regulatory protein, positively controls catecholamine secretion in PC12D cells. FEBS Lett. 530, 94–98 (2002)

    Article  CAS  Google Scholar 

  24. Lewis, B. P., Shih, I. H., Jones-Rhoades, M. W., Bartel, D. P. & Burge, C. B. Prediction of mammalian microRNA targets. Cell 115, 787–798 (2003)

    Article  CAS  Google Scholar 

  25. Lagos-Quintana, M. et al. Identification of tissue-specific microRNAs from mouse. Curr. Biol. 12, 735–739 (2002)

    Article  CAS  Google Scholar 

  26. Minami, K. et al. Insulin secretion and differential gene expression in glucose-responsive and -unresponsive MIN6 sublines. Am. J. Physiol. Endocrinol. Metab. 279, E773–E781 (2000)

    Article  CAS  Google Scholar 

  27. Shih, D. Q. et al. Profound defects in pancreatic beta-cell function in mice with combined heterozygous mutations in Pdx-1, Hnf-1α, and Hnf-3β. Proc. Natl Acad. Sci. USA 99, 3818–3823 (2002)

    Article  ADS  CAS  Google Scholar 

  28. Eliasson, L. et al. SUR1 regulates PKA-independent cAMP-induced granule priming in mouse pancreatic B-cells. J. Gen. Physiol. 121, 181–197 (2003)

    Article  CAS  Google Scholar 

  29. Olofsson, C. S. et al. Fast insulin secretion reflects exocytosis of docked granules in mouse pancreatic B-cells. Pflugers Arch. 444, 43–51 (2002)

    Article  CAS  Google Scholar 

  30. Zuker, M. Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Res. 31, 3406–3415 (2003)

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank K. Borglid, J. Chen, J. Galvanovskis, M. Lagos-Quintana, M. Landthaler, A. Lingqvist, G. Meister, B. M. Nilsson, A. Wendt and C. Wolfrum for advice and technical assistance. This work was supported by an unrestricted grant from Bristol Myers Squibb, the Juvenile Diabetes Research Foundation, the Deutsche Forschungsgemeinschaft and grants from the Swedish Research Council, the Swedish Diabetes Association, the Göran Gustafsson Stiftelse for Natural Sciences and Medicine and the Swedish Strategic Research Foundation (SSF).

Author information

Authors and Affiliations

  1. Laboratory of Metabolic Diseases and Laboratory of RNA Molecular Biology, The Rockefeller University, 1230 York Avenue, New York, New York, 10021, USA

    Matthew N. Poy, Jan Krutzfeldt, Satoru Kuwajima, Sébastien Pfeffer, Thomas Tuschl & Markus Stoffel

  2. Department of Physiological Sciences, Lund University, SE-221 84, Lund, Sweden

    Lena Eliasson, Xiaosong Ma, Patrick E. MacDonald & Patrik Rorsman

  3. Department of Biology, Biology & Mathematics, New York University, New York, New York, 10003, USA

    Nikolaus Rajewsky

  4. Oxford Centre for Diabetes, Endocrinology and Metabolism, University of Oxford, Churchill Hospital, OX3 7LJ, Oxford, UK

    Patrik Rorsman

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  1. Matthew N. Poy
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Correspondence to Markus Stoffel.

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Supplementary information

Supplementary Figure 1

Oligoribonucleotide 2’-O-me-375 anneals to endogenous miR-375 in MIN6 cells. Northern blot of MIN6 cells that were transfected with either 2’O-me-eGFP (control) or 2’-O-me-375. (PDF 84 kb)

Supplementary Figure 2

Increased [ATP]i levels in MIN6 cells infected with Ad-375 compared to Ad-eGFP. Total ATP measurements in MIN6 cells that were transfected with Ad-375 or Ad-eGFP. (PDF 42 kb)

Supplementary Figure 3

No apparent effects of miR-375 overexpression on submembrane actin network. Structural analysis using fluorescence microscopy of the submembrane actin network in MIN6 cells and primary β-cells that were infected with Ad-eGFP or Ad-375. (PDF 717 kb)

Supplementary Figure 4

Increased number of docked granules in Ad-375 infected β-cells. Electron micrographs of β-cells infected with Ad-eGFP or Ad-375 and quantitative analysis of granule density. (PDF 855 kb)

Supplementary Table 1

Mature and precursor sequences of mouse (mmu-miR) and human (hsa-miR) novel miRNAs, identified in mouse pancreatic α- and β-cell lines TC1 and MIN6, respectively. (DOC 182 kb)

Supplementary Table 2

Foldback precursor sequences of novel mouse miRNAs, identified in mouse pancreatic β-cell line MIN6. (DOC 24 kb)

Supplementary Table 3

Similar [Ca++]i in primary β-cells and MIN6 cells infected with Ad-GFP and Ad-375. Ca2+-measurements in primary β-cells and MIN6 cells infected with Ad-GFP and Ad-375 in response to 25 mM glucose and to 30 mM K+. (DOC 23 kb)

Supplementary Table 4

Similar [Ca++]i in response to glucose and K+ in primary b-cells and MIN6 cells infected with control siRNAs, si-375 and si-Mtpn. Ca2+-measurements in MIN6 cells co-transfected with CMV-eGFP and si-ApoM (control), si-375 or si-Mtpn in response to glucose K+. (DOC 20 kb)

Supplementary Methods

Description of methods used to identify miR-375 targets. (DOC 24 kb)

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Poy, M., Eliasson, L., Krutzfeldt, J. et al. A pancreatic islet-specific microRNA regulates insulin secretion. Nature 432, 226–230 (2004). https://doi.org/10.1038/nature03076

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