Clarification of characteristics, functions and regulation of proteoglycans synthesized by pericytes

1998 Fiscal Year Final Research Report Summary

• Project/Area Number: 09672260
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biological pharmacy
• Research Institution: Hokuriku University
• Principal Investigator: KAJI Toshiyuki Hokuriku University, Faculty of Pharmaceutical Sciences, Associate Professor, 薬学部, 助教授 (90204388)
• Co-Investigator(Kenkyū-buntansha): MIYAJIMA Sawako Hokuriku University, Faculty of Pharmaceutical Sciences, Research Associate, 薬学部, 助手 (30278139) ; FUJIWARA Yasuyuki Hokuriku University, Faculty of Pharmaceutical Sciences, Research Associate, 薬学部, 助手 (40247482) ; YAMAMOTO Chika Hokuriku University, Faculty of Pharmaceutical Sciences, Research Associate, 薬学部, 助手 (70230571)
• Project Period (FY): 1997 – 1998
• Keywords: Pericytes / Proteoglycan / Versican / Biglycan / Decorin / Perlecan / Core protein / Vascular

Research Abstract

Proteoglycans synthesized by pericytes were characterized by biochemical techniques. Bovine retinal pericytes were cultured and metabolically labeled with [^<35>S] sutfate or ^<35>S amino acids. The proteoglycans labeled with [^<35>S] sulfate were separated into three peaks by DEAE-Sephacel ion exchange chromatography. Sepharose CL-6B chromatography showed , that the peak with the lowest charge density is composed of heparan sulfate proteogly cans whereas the higher two peaks are composed of chondroitin/dermatan sulfate proteoglycans (CS/DS PGs). Since pericytes predominantly synthesized CS/DS PGs, the CS/DS PGs isolated by DEAE-Sephacel chromatography were analyzed by Sepharose CL-4B and Sepharose CL-2B gel chromatography and SDS-PAGE followed by Western blot analysis. It was revealed that pericytes synthesize two subclass of CS/DS PGs ; one is highly charged versican molecule with 45 kDa glycosaminoglycan chains and the other is low charged biglycan and decorin molecules with 45 kDa glycosaminoglycan chains. Although our preliminary study suggested that the synthesis of versican, biglycan, decorin and perlecan can be regulated by growth factors and cytokines including platelet-derived growth factor, basic fibroblast growth factor, transforming growth factor-beta and tumor necrosis factor-alpha, the structural changes of each proteogly can molecule remain to be elucidated.