The purpose of these studies was to characterize the development of H+ secretion by cultured inner medullary collecting duct (IMCD) cells. Differentiated IMCD cells derived from rat papillae develop in culture N-ethylmaleimide (NEM)- and dicyclohexylcarbodiimide (DCCD)-sensitive Na+-independent H+ pump, but showed substantial heterogeneity. Cells with pH(i) ≥ 7.1, measured by 2',7'-biscarboxyethyl-5(6)-carboxyfluorescein (BCECF), showed more H+ pumping than did those with pH(i) < 7.1. Quiescent cells in domes and in the center of growing nests often showed active H+ pumping activity, but rarely exhibited Na+-H+ exchange, whereas dividing cells at the periphery of the nests showed Na+-independent 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS)-sensitive Cl--HCO3- exchange that functioned best when pH(i) was ≥ 7.1, and homogenates of IMCD cells expressed carbonic anhydrase (CA) activity and CA II mRNA. A subgroup of IMCD cells avidly bound fluorescent peanut agglutinin (PNA); this binding became more extensive with time in culture. The PNA-labeled cells showed increased BCECF uptake, higher pH(i), and more H+ pumping activity than did cells not binding PNA. They could be isolated by gel-immobilized PNA and in culture showed more CA activity and H+ pumping activity than unselected monolayers. The percentage of cells binding PNA correlated well with CA activity, indicating that this subgroup is better specialized to secrete H+. The appearance of PNA binding sites and the development of high CA activity may be characteristic of IMCD cell differentiation in culture.
|Original language||English (US)|
|Journal||American Journal of Physiology - Renal Fluid and Electrolyte Physiology|
|Issue number||3 (26/3)|
|State||Published - 1989|
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