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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
Article

Abstract

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.

Keywords

Db/db islets β-cell dedifferentiation Network analysis 

Abbreviations

MIN6

Mouse insulinoma 6

Vdr

Vitamin D receptor

Ins1

Insulin 1

Ins2

Insulin 2

Glut2

Glucose transporter 2

Pdx1

Pancreatic and duodenal homebox1

Pax6

Paired Box 6

Gcg

Glucagon

Gck

Glucokinase

MafA

V-Maf avian musculoaponeurotic fibrosarcoma oncogene homologue

βCD

β-cell dedifferentiation

T2D

Type 2 diabetes

−/− mice

Wild-type mice

db/− mice

Heterozygous mice

db/db mice

Homozygous mice

Gc

Vitamin D binding protein

Aldh1a3

Aldehyde dehydrogenase

Serpina7

Thyroxin-binding globulin

Mir6263

MicroRNA 6236

Irs1

Insulin receptor substrate 1

Slc2a2

Solute carrier family 2 (facilitated glucose transporter)

Stat6

Signal transducer and activator of transcription

Ffar

Free fatty acid receptor

Igf1r

Insulin-like growth factor 1 receptor

Prkab2

Protein kinase AMP-activated non-catalytic beta-2

Trib

Tribbles

Npas4

Neuronal PAS domain protein 4

Rasgrf1

Ras protein-specific guanine nucleotide releasing factor 1

Htt

Huntington gene

Pparg

Peroxisome proliferator-activated receptor

Fosl1

Fos-like antigen 1

Gipr

Gastric inhibitory polypeptide receptor

Nr4a1

Nuclear receptor subfamily 4

Pclo

Piccolo

Ptrf

Cavin1

Ucn3

Urocortin III

Slc2a2

Protein phosphatase 1

papss2

Prime phosphoadenosine 5 prime phosphosulfate synthase 2

Gnas

Gnas complex locus

Ero1b

Endoplasmic reticulum oxidorectin 1-like beta

Notes

Acknowledgements

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.

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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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