{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,8]],"date-time":"2026-05-08T20:58:22Z","timestamp":1778273902900,"version":"3.51.4"},"reference-count":62,"publisher":"American Physiological Society","issue":"9","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["American Journal of Physiology-Renal Physiology"],"published-print":{"date-parts":[[2012,11,1]]},"abstract":"<jats:p>The thick ascending limb of the loop of Henle (THAL) reabsorbs \u223c30% of the filtered NaCl in a process mediated by the apical Na-K-2Cl cotransporter NKCC2. Stimulation of \u03b2-adrenergic receptors in the THAL enhances NaCl reabsorption and increases intracellular cAMP. We found that intracellular cAMP stimulates NKCC2 trafficking to the apical membrane via protein kinase A (PKA). Several cAMP-specific phosphodiesterases (PDE) have been identified in rat THALs, and PDE4 decreases cAMP generated by \u03b2-adrenergic stimulation in other cells. However, it is not known whether \u03b2-adrenergic receptors activation stimulates NKCC2 trafficking. Thus we hypothesized that \u03b2-adrenergic receptor stimulation enhances THAL apical membrane NKCC2 expression via the PKA pathway and PDE4 blunts this effect. THAL suspensions were obtained from Sprague-Dawley rats, and surface NKCC2 expression was measured by surface biotinylation and Western blot. Incubation of THALs with the \u03b2-adrenergic receptor agonist isoproterenol at 0.5 and 1.0 \u03bcM increased surface NKCC2 by 17 \u00b1 1 and 29 \u00b1 5% respectively ( P &lt; 0.05). Preventing cAMP degradation with 3-isobutyl-methylxanthine (IBMX; a nonselective phosphodiesterase inhibitor) enhanced isoproterenol-stimulated surface NKCC2 expression to 51 \u00b1 7% ( P &lt; 0.05 vs. isoproterenol). The \u03b2-adrenergic receptor antagonist propranolol or the PKA inhibitor H-89 completely blocked isoproterenol + IBMX-induced increase on surface NKCC2, while propranolol or H-89 alone had no effect. Selective inhibition of PDE4 with rolipram (20 \u03bcM) potentiated the effect of isoproterenol on surface NKCC2 and increased cAMP levels. We concluded that \u03b2-adrenergic receptor stimulation enhances surface NKCC2 expression in the THALs via PKA and PDE4 blunts this effect.<\/jats:p>","DOI":"10.1152\/ajprenal.00019.2012","type":"journal-article","created":{"date-parts":[[2012,8,30]],"date-time":"2012-08-30T01:08:24Z","timestamp":1346288904000},"page":"F1307-F1314","source":"Crossref","is-referenced-by-count":21,"title":["\u03b2-Adrenergic receptor stimulation increases surface NKCC2 expression in rat thick ascending limbs in a process inhibited by phosphodiesterase 4"],"prefix":"10.1152","volume":"303","author":[{"given":"Mohammed Z.","family":"Haque","sequence":"first","affiliation":[{"name":"Hypertension and Vascular Research, Department of Internal Medicine, Henry Ford Hospital, Detroit, Michigan; and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paulo S.","family":"Caceres","sequence":"additional","affiliation":[{"name":"Hypertension and Vascular Research, Department of Internal Medicine, Henry 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