Qualitative and quantitative analyses in sulfated glycosaminoglycans, chondroitin sulfate/dermatan sulfate, during 3 T3-L1 adipocytes differentiation

Anim Sci J. 2023 Jan-Dec;94(1):e13894. doi: 10.1111/asj.13894.

Abstract

Chondroitin sulfate/dermatan sulfate (CS/DS) is a member of glycosaminoglycans (GAGs) found in animal tissues. Major CS/DS subclasses, O, A, C, D, and E units, exist based on the sulfation pattern in d-glucuronic acid (GlcA) and N-acetyl-d-galactosamine repeating units. DS is formed when GlcA is epimerized into l-iduronic acid. Our study aimed to analyze the CS/DS profile in 3 T3-L1 cells before and after adipogenic induction. CS/DS contents, molecular weight (Mw), and sulfation pattern were analyzed by using high-performance liquid chromatography. CS/DS synthesis- and sulfotransferase-related genes were analyzed by reverse transcription real-time PCR. CS/DS amount was significantly decreased in the differentiated (DI) group compared to the non-differentiated (ND) group, along with a lower expression of CS biosynthesis-related genes, chondroitin sulfate N-acetylgalactosaminyltransferase 1 and 2, as well as chondroitin polymerizing factor. GAGs in the DI group also showed lower Mw than those of ND. Furthermore, the A unit was the major CS/DS in both groups, with a proportionally higher CS-A in the DI group. This was consistent with the expression of carbohydrate sulfotransferase 12 that encodes chondroitin 4-O-sulfotransferase, for CS-A formation. These qualitative and quantitative changes in CS/DS and CS/DS-synthases before and after adipocyte differentiation reveal valuable insights into adipocyte development.

Keywords: 3 T3-L1; adipocytes; adipogenesis; chondroitin sulfate; glycosaminoglycans.

MeSH terms

  • Animals
  • Cell Differentiation
  • Chondroitin Sulfates* / analysis
  • Chondroitin Sulfates* / chemistry
  • Chondroitin Sulfates* / metabolism
  • Dermatan Sulfate* / analysis
  • Dermatan Sulfate* / metabolism
  • Dermatan Sulfate* / pharmacology
  • Glycosaminoglycans / metabolism
  • Sulfotransferases / genetics
  • Sulfotransferases / metabolism

Substances

  • Chondroitin Sulfates
  • A73025
  • Dermatan Sulfate
  • Glycosaminoglycans
  • Sulfotransferases