Endocrine Pathology

, Volume 29, Issue 3, pp 207–221 | Cite as

RNA-Seq Analysis of Islets to Characterise the Dedifferentiation in Type 2 Diabetes Model Mice db/db

  • Abraham Neelankal John
  • Ramesh Ram
  • Fang-Xu Jiang


Type 2 diabetes (T2D) is a global health issue and dedifferentiation plays underlying causes in the pathophysiology of T2D; however, there is a lack of understanding in the mechanism. Dedifferentiation results from the loss of function of pancreatic β-cells alongside a reduction in essential transcription factors under various physiological stressors. Our study aimed to establish db/db as an animal model for dedifferentiation by using RNA sequencing to compare the gene expression profile in islets isolated from wild-type, db/+ and db/db mice, and qPCR was performed to validate those significant genes. A reduction in both insulin secretion and the expression of Ins1, Ins2, Glut2, Pdx1 and MafA was indicative of dedifferentiation in db/db islets. A comparison of the db/+ and the wild-type islets indicated a reduction in insulin secretion perhaps related to the decreased Mt1. A significant reduction in both Rn45s and Mir6236 was identified in db/+ compared to wild-type islets, which may be indicative of pre-diabetic state. A further significant reduction in RasGRF1, Igf1R and Htt was also identified in dedifferentiated db/db islets. Molecular characterisation of the db/db islets was performed via Ingenuity analysis which identified highly significant genes that may represent new molecular markers of dedifferentiation.


Db/db islets β-cell dedifferentiation Network analysis 



Mouse insulinoma 6


Vitamin D receptor


Insulin 1


Insulin 2


Glucose transporter 2


Pancreatic and duodenal homebox1


Paired Box 6






V-Maf avian musculoaponeurotic fibrosarcoma oncogene homologue


β-cell dedifferentiation


Type 2 diabetes

−/− mice

Wild-type mice

db/− mice

Heterozygous mice

db/db mice

Homozygous mice


Vitamin D binding protein


Aldehyde dehydrogenase


Thyroxin-binding globulin


MicroRNA 6236


Insulin receptor substrate 1


Solute carrier family 2 (facilitated glucose transporter)


Signal transducer and activator of transcription


Free fatty acid receptor


Insulin-like growth factor 1 receptor


Protein kinase AMP-activated non-catalytic beta-2




Neuronal PAS domain protein 4


Ras protein-specific guanine nucleotide releasing factor 1


Huntington gene


Peroxisome proliferator-activated receptor


Fos-like antigen 1


Gastric inhibitory polypeptide receptor


Nuclear receptor subfamily 4






Urocortin III


Protein phosphatase 1


Prime phosphoadenosine 5 prime phosphosulfate synthase 2


Gnas complex locus


Endoplasmic reticulum oxidorectin 1-like beta



We also thank the support of Diabetes Research (WA). Thanks to Dr. Angela Abraham and Leah Simmons for their suggestions.

Authors’ Contributions

ANJ wrote the manuscript, all the laboratory works and part of the bioinformatic analysis; RR performed the bioinformatics analysis and FXJ supervised this work.

Funding information

We thank Telethon and the Perth Child Research Fund and the University of Western Australia for their financial support.

Compliance with Ethical Standards

The Animal Resource Centre Ethics Committee, Murdoch University approved the use of mice for islet purification of wild-type, db/+ and db/db.

Conflict of Interest

The authors declare that they have no conflict of interest.


  1. 1.
    Matthews, D.R., et al., UKPDS 26: Sulphonylurea failure in non-insulin-dependent diabetic patients over six years. UK Prospective Diabetes Study (UKPDS) Group. Diabet Med, 1998. 15(4): p. 297–303.CrossRefPubMedGoogle Scholar
  2. 2.
    Neelankal John, A., G. Morahan, and F.X. Jiang, Incomplete re-expression of neuroendocrine progenitor/stem cell markers is a key feature of beta-cell dedifferentiation. J Neuroendocrinol, 2017. 29(1).
  3. 3.
    Venardos, K., et al., The PKD inhibitor CID755673 enhances cardiac function in diabetic db/db mice. PLoS One, 2015. 10(3): p. e0120934.Google Scholar
  4. 4.
    Belke, D.D. and D.L. Severson, Diabetes in mice with monogenic obesity: the db/db mouse and its use in the study of cardiac consequences. Methods Mol Biol, 2012. 933: p. 47–57.PubMedGoogle Scholar
  5. 5.
    Harris, R.B., et al., Metabolic responses to leptin in obese db/db mice are strain dependent. Am J Physiol Regul Integr Comp Physiol, 2001. 281(1): p. R115–R132.CrossRefPubMedGoogle Scholar
  6. 6.
    Kobayashi, K., et al., The db/db mouse, a model for diabetic dyslipidemia: molecular characterization and effects of Western diet feeding. Metabolism, 2000. 49(1): p. 22–31.CrossRefPubMedGoogle Scholar
  7. 7.
    Sticco, S.L., Post-intubation croup. CRNA, 1995. 6(3): p. 143–144.PubMedGoogle Scholar
  8. 8.
    Bogdanov, P., et al., The db/db mouse: a useful model for the study of diabetic retinal neurodegeneration. PLoS One, 2014. 9(5): p. e97302.Google Scholar
  9. 9.
    Zhao, G., et al., Delayed wound healing in diabetic (db/db) mice with Pseudomonas aeruginosa biofilm challenge: a model for the study of chronic wounds. Wound Repair Regen, 2010. 18(5): p. 467–477.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Greer, J.J., D.P. Ware, and D.J. Lefer, Myocardial infarction and heart failure in the db/db diabetic mouse. Am J Physiol Heart Circ Physiol, 2006. 290(1): p. H146–H153.CrossRefPubMedGoogle Scholar
  11. 11.
    Garris, B.L., et al., Hypophyseal lipoapoptosis: diabetes (db/db) mutation-associated cytolipidemia promotes pituitary cellular disruption and dysfunction. Pituitary, 2004. 7(1): p. 5–14.CrossRefPubMedGoogle Scholar
  12. 12.
    Garris, D.R., Diabetes (db/db) mutation-induced endometrial epithelial lipoapoptosis: ultrastructural and cytochemical analysis of reproductive tract atrophy. Reprod Biol Endocrinol, 2005. 3: p. 15.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Talchai, C., et al., Pancreatic beta cell dedifferentiation as a mechanism of diabetic beta cell failure. Cell, 2012. 150(6): p. 1223–1234.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Patti, M.E., P. Li, and A.B. Goldfine, Insulin response to oral stimuli and glucose effectiveness increased in neuroglycopenia following gastric bypass. Obesity (Silver Spring), 2015. 23(4): p. 798–807.CrossRefGoogle Scholar
  15. 15.
    Mueckler, M. and B. Thorens, The SLC2 (GLUT) family of membrane transporters. Mol Aspects Med, 2013. 34(2–3): p. 121–138.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Neelankal John, A., et al., Vitamin D receptor-targeted treatment to prevent pathological dedifferentiation of pancreatic beta cells under hyperglycaemic stress. Diabetes Metab, 2017.Google Scholar
  17. 17.
    Agouridis, A.P., M.S. Kostapanos, and M.S. Elisaf, Statins and their increased risk of inducing diabetes. Expert Opin Drug Saf, 2015. 14(12): p. 1835–1844. CrossRefPubMedGoogle Scholar
  18. 18.
    Neelankal John, A. and F.X. Jiang, An overview of type 2 diabetes and importance of vitamin D3-vitamin D receptor interaction in pancreatic beta-cells. J Diabetes Complications, 2017.Google Scholar
  19. 19.
    Chang, I., et al., Role of calcium in pancreatic islet cell death by IFN-gamma/TNF-alpha. J Immunol, 2004. 172(11): p. 7008–7014.CrossRefPubMedGoogle Scholar
  20. 20.
    Sandler, S., et al., Interleukin-6 affects insulin secretion and glucose metabolism of rat pancreatic islets in vitro. Endocrinology, 1990. 126(2): p. 1288–1294.CrossRefPubMedGoogle Scholar
  21. 21.
    Ito, E., et al., PPAR-gamma overexpression selectively suppresses insulin secretory capacity in isolated pancreatic islets through induction of UCP-2 protein. Biochem Biophys Res Commun, 2004. 324(2): p. 810–814.CrossRefPubMedGoogle Scholar
  22. 22.
    Peschke, E., et al., Receptor (MT(1)) mediated influence of melatonin on cAMP concentration and insulin secretion of rat insulinoma cells INS-1. J Pineal Res, 2002. 33(2): p. 63–71.CrossRefPubMedGoogle Scholar
  23. 23.
    Hamidi, T., et al., Nuclear protein 1 promotes pancreatic cancer development and protects cells from stress by inhibiting apoptosis. J Clin Invest, 2012. 122(6): p. 2092–2103.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Patil, D.P., et al., m(6)A RNA methylation promotes XIST-mediated transcriptional repression. Nature, 2016. 537(7620): p. 369–373.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Lindahl, M., et al., MANF is indispensable for the proliferation and survival of pancreatic beta cells. Cell Rep, 2014. 7(2): p. 366–375.CrossRefPubMedGoogle Scholar
  26. 26.
    Cape, A., et al., Loss of huntingtin-associated protein 1 impairs insulin secretion from pancreatic beta-cells. Cell Mol Life Sci, 2012. 69(8): p. 1305–1317.CrossRefPubMedGoogle Scholar
  27. 27.
    Font de Mora, J., et al., Ras-GRF1 signaling is required for normal beta-cell development and glucose homeostasis. EMBO J, 2003. 22(12): p. 3039–3049.
  28. 28.
    Kim-Muller, J.Y., et al., Aldehyde dehydrogenase 1a3 defines a subset of failing pancreatic beta cells in diabetic mice. Nat Commun, 2016. 7: p. 12631.CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Kushch, N.L., et al., [Clinico-immunological indices, lipid peroxidation and contents of middle mass molecules in young children with different course of suppurative diseases of the lungs and pleura]. Klin Khir, 1990(6): p. 18–20.Google Scholar
  30. 30.
    Rak, M., et al., Mitochondrial cytochrome c oxidase deficiency. Clin Sci (Lond), 2016. 130(6): p. 393–407.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Harry Perkins Institute of Medical Research, Centre for Medical ResearchUniversity of Western AustraliaNedlandsAustralia
  2. 2.School of Medicine And PharmacologyUniversity of Western AustraliaCarwleyAustralia
  3. 3.Islet Cell Development ProgramHarry Perkins Institute of Medical ResearchPerth WesternAustralia

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