Ontogenic regulation of folate transport across rat jejunal brush-border membrane

Abstract

Folate is an essential micronutrient that in mammals must be obtained from exogenous sources via intestinal absorption. Previous studies from our laboratory and others have demonstrated that folate absorption from the small intestine is mediated via the reduced-folate carrier (RFC). The goal of this study was to determine whether the initial step of folate uptake by intestinal epithelial cells, i.e., transport across the brush-border membrane (BBM) of the polarized enterocytes, is ontogenically regulated, and if so, to determine the molecular mechanism involved. Purified BBM vesicles (BBMV) isolated from suckling, weanling, and adult rats were used in this study. The initial rate of carrier-mediated uptake of a physiological concentration of folic acid (0.1 muM) by jejunal BBMV was found to be significantly (P < 0.01) higher in suckling compared with weanling rats, which was, in turn, significantly (P < 0.01) higher than that in adult rats. This decline in carrier-mediated folate uptake with maturation was found to be mediated via a decrease in the maximum velocity of the folate uptake process (6.55 +/- 0.87, 2.16 +/- 0.10, 0.90 +/- 0.16 pmol.mg protein-1.10 s-1 for suckling, weanling, and adult rats, respectively), with no changes in its apparent Km. Western blot analysis of BBM protein and real-time PCR showed RFC protein and mRNA levels, respectively, to be significantly (P < 0.01 for both) higher in suckling compared with weanling rats, which were in turn significantly (P < 0.01 for both) higher than that in adult rats. These changes were found by nuclear run-on assay to be associated with a parallel decline in the RFC transcriptional rate in jejunal epithelia with maturation. In situ hybridization showed a similar pattern of RFC message distribution along crypt/villus axis in suckling and adult rat jejunum. These results demonstrate for the first time that folate transport across the intestinal BBM is under ontogenic regulation during early stages of life and that this regulation involves a transcriptional regulatory mechanism(s).