Development of H+ secretion by cultured renal inner medullary collecting duct cells

L. P. Brion, J. H. Schwartz, Herbert M. Lachman, B. J. Zavilowitz, G. J. Schwartz

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Abstract

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 languageEnglish (US)
JournalAmerican Journal of Physiology - Renal Fluid and Electrolyte Physiology
Volume257
Issue number3
StatePublished - 1989

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Peanut Agglutinin
Kidney
Carbonic Anhydrases
Carbonic Anhydrase II
Dicyclohexylcarbodiimide
Proton Pumps
Ethylmaleimide
Cell Differentiation
Gels
Binding Sites
Messenger RNA
Acids

ASJC Scopus subject areas

  • Physiology

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Development of H+ secretion by cultured renal inner medullary collecting duct cells. / Brion, L. P.; Schwartz, J. H.; Lachman, Herbert M.; Zavilowitz, B. J.; Schwartz, G. J.

In: American Journal of Physiology - Renal Fluid and Electrolyte Physiology, Vol. 257, No. 3, 1989.

Research output: Contribution to journalArticle

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