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Vitamin C and collagen IV formation in fibroblasts

by Burl, Lana R., M.S., STATE UNIVERSITY OF NEW YORK AT BUFFALO, 2007, 65 pages; 1440040

Abstract:

Collagen IV (Col4) is a major component of vascular basement membranes (BM). Mice expressing mutated Col4 died shortly after birth with cerebral hemorrhage. Mice lacking the ubiquitous vitamin C (Asc) transporter SVCT2 suffered similar perinatal death. Col4 maturation depends on Asc, and we hypothesized that SVCT2 is essential for normal cell growth and Col4 formation. Mouse embryonic fibroblasts of three genotypes, SVCT2+/+, SVCT2+/-, SVCT2-/-, were cultured. Asc transport throughout subculturing demonstrated genotype preservation. SVCT2-/- had more aggressive proliferation, achieving higher saturation densities in less time than SVCT2+/+ and SVCT2+/-. Ascorbate-2-phosphate supplementation (AscPO4 ) increased protein in SVCT2+/+ and SVCT2+/-, but not SVCT2-/-. In western blotting, AscPO4 tended to decrease soluble Col4 in SVCT2+/+. Confocal immunocytochemistry revealed enhanced nuclear localization of Col4 in SVCT2+/+ upon AscPO4 . AscPO4 appeared to increase se Col4 to actin ratio in SVCT2+/+ but not in SVCT2-/-. Lack of SVCT2 led to altered fibroblast growth and Col4 formation.

Advisor:	Kuo, Shiu-Ming
School:	STATE UNIVERSITY OF NEW YORK AT BUFFALO
Source:	MAI 45/02, p. , Apr 2007
Source Type:	M.S.
Subjects:	Cellular biology; Biochemistry; Nutrition
Publication Number:	1440040

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