Ion Transport during Growth and Differentiation

Abstract

The major function of the adult colon is to reabsorb fluid from the chyme. This ability to conserve salt and water is especially important in newborns, where reserves are small and diarrhea is frequent. Although much is known about regulation of Cl- transport in the adult colon, postnatal changes in electrolyte transport are not well characterized. We have established an in vitro model to study colonic epithelial cells (colonocytes) at different stages of development. Primary cultures were isolated from newborn, weanling, and adult rabbit colon and properties such as growth and Cl- transport characterized. The isolation procedure yielded a crypt-enriched population of cells, and the cell yield per gram mucosa increased with age. The colonocytes also showed an age-related decrease in attachment to extracellular matrix, with maximum attachment seen with Matrigel and collagen IV. The crypt enrichment was confirmed by demonstrating that the cell population was capable of transporting Cl-, which was stimulated by agents such as forskolin and phorbol esters at all ages. Agents that increased intracellular cGMP, however, did not increase Cl- transport at any age. It was interesting to observe that the secondary bile acid, taurodeoxycholate, stimulated Cl- transport only in the adult but not newborn or weanling distal colonocytes. We have demonstrated that rabbit distal colonocytes can be kept viable in culture and transport Cl- at all ages. However, the regulation of Cl- transport changes during ontogeny and depends on the signaling pathway.