Differential expression of genes identified by suppression subtractive hybridization in liver and adipose tissue of gerbils with diabetes

PLoS One. 2018 Feb 2;13(2):e0191212. doi: 10.1371/journal.pone.0191212. eCollection 2018.

Abstract

Objectives: We aimed at identifying genes related to hereditary type 2 diabetes expressed in the liver and the adipose tissue of spontaneous diabetic gerbils using suppression subtractive hybridization (SSH) screening.

Methods: Two gerbil littermates, one with high and the other with normal blood glucose level, from our previously bred spontaneous diabetic gerbil strain were used in this study. To identify differentially expressed genes in the liver and the adipose tissue, mRNA from these tissues was extracted and SSH libraries were constructed for screening. After sequencing and BLAST analyzing, up or down-regulated genes possibly involved in metabolism and diabetes were selected, and their expression levels in diabetic gerbils and normal controls were analyzed using quantitative RT-PCR and Western blotting.

Results: A total of 4 SSH libraries were prepared from the liver and the adipose tissue of gerbils. There are 95 up or down-regulated genes were identified to be involved in metabolism, oxidoreduction, RNA binding, cell proliferation, and differentiation or other function. Expression of 17 genes most possibly associated with diabetes was analyzed and seven genes (Sardh, Slc39a7, Pfn1, Arg1, Cth, Sod1 and P4hb) in the liver and one gene (Fabp4) in the adipose tissue were identified that were significantly differentially expressed between diabetic gerbils and control animals.

Conclusions: We identified eight genes associated with type 2 diabetes from the liver and the adipose tissue of gerbils via SSH screening. These findings provide further insights into the molecular mechanisms of diabetes and imply the value of our spontaneous diabetic gerbil strain as a diabetes model.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Adipose Tissue / metabolism
  • Animals
  • Blotting, Western
  • Diabetes Mellitus, Experimental / genetics
  • Diabetes Mellitus, Experimental / metabolism
  • Diabetes Mellitus, Type 2 / genetics*
  • Diabetes Mellitus, Type 2 / metabolism
  • Disease Models, Animal
  • Gene Expression Profiling
  • Gene Library
  • Gerbillinae / genetics*
  • Gerbillinae / metabolism
  • Liver / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Real-Time Polymerase Chain Reaction
  • Subtractive Hybridization Techniques

Substances

  • RNA, Messenger

Grants and funding

This study was funded by the National Science Foundation of China Nos. 31272393 and 31572341 Xiaoyan Du, No 31572348 Changlong Li, No 31402027 Meng Guo, the website is http://www.nsfc.gov.cn/, Key Projects in the National Science & Technology Pillar Program (No. 2015BAI09B01) and the Beijing Natural Science Foundation (No.7141002), Zhenwen Chen, the website is http://bjnsf.bjkw.gov.cn/.