Na+-K+-ATPase-mediated basolateral rubidium uptake in the maturing rabbit cortical collecting duct

Alexandru R. Constantinescu, Jerome C. Lane, John Mak, Beth Zavilowitz, Lisa M. Satlin*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

Within the renal cortical collecting duct (CCD), transepithelial Na+ absorption and K+ secretion are linked to basolateral Na+-K+-ATPase activity. Our purpose was to examine the developmental changes in basolateral Na+-K+-ATPase-mediated 86rubidium (Rb) uptake, its inhibitor sensitivity and relationship to pump hydrolytic activity and Na+ transport. Multiple CCDs (~6 mm) from maturing rabbits were affixed to coverslips, preincubated at 37°C for 10 min (±1-2.5 mM ouabain or 10 or 100 μM Schering-28080, an inhibitor of H+-K+-ATPase), and then transferred to prewarmed incubation solution containing tracer amounts of 86Rb (±inhibitors). After 1 min at 37°C, tubular samples were rinsed and permeabilized and isotope counts were measured to calculate basolateral Rb uptake. Ouabain-inhibitable Rb uptake, an index of basolateral Na+-K+ pump activity, increased ~3-fold during the 1st 8 wk of postnatal life (P < 0.03). The ~2-fold increase in absolute rate of Rb uptake between 1 and 6 wk (2.64 ± 0.45 to 5.02 ± 0.32 pmol·min-1·mm-1) did not reach statistical significance. The rate of basolateral Rb uptake increased further after the 6th wk of life to 7.29 ± 0.53 pmol·min-1·mm-1 in adult animals (P < 0.03 vs. 6 wk). Schering-28080 failed to inhibit Rb uptake, implying that functional H+-K+-ATPase is absent at the basolateral membrane. Na+-K+-ATPase hydrolytic activity, determined by using a microassay that measured inorganic phosphate release from [γ32P]ATP under maximum velocity (V(max)) conditions, also increased in the differentiating CCD (from 316.2 ± 44.4 pmol·h-1·mm-1 at 2 wk to 555.9 ± 105.1 at 4 wk to 789.7 ± 145.0 at 6 wk; r = 1.0 by linear regression analysis; P < 0.005). The parallel ~2.5-fold increases in Na+-K+-ATPase activity and ouabain-sensitive Rb uptake between 2- and 6-wk postnatal age suggest that the developmental increase in basolateral transport capacity is due predominantly to an increase in enzyme abundance. The signals mediating the developmental increase in Na+-K+-ATPase activity in the CCD remain to be defined.

Original languageEnglish (US)
Pages (from-to)F1161-F1168
JournalAmerican Journal of Physiology - Renal Physiology
Volume279
Issue number6 48-6
DOIs
StatePublished - 2000

Keywords

  • Microassay
  • Ontogeny
  • Ouabain
  • Schering-28080
  • Transepithelial sodium transport

ASJC Scopus subject areas

  • Physiology
  • Urology

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