Vitamin C and collagen IV formation in fibroblasts
Burl, Lana R
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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 (AscPO 4 ) increased protein in SVCT2+/+ and SVCT2+/-, but not SVCT2-/-. In western blotting, AscPO 4 tended to decrease soluble Col4 in SVCT2+/+. Confocal immunocytochemistry revealed enhanced nuclear localization of Col4 in SVCT2+/+ upon AscPO 4 . AscPO 4 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.