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  • Title: Murine vasa recta pericyte chloride conductance is controlled by calcium, depolarization, and kinase activity.
    Author: Lin H, Pallone TL, Cao C.
    Journal: Am J Physiol Regul Integr Comp Physiol; 2010 Nov; 299(5):R1317-25. PubMed ID: 20686172.
    Abstract:
    We used the whole cell patch-clamp technique to investigate the regulation of descending vasa recta (DVR) pericyte Ca(2+)-dependent Cl(-) currents (CaCC) by cytoplasmic Ca(2+) concentration ([Ca](CYT)), voltage, and kinase activity. Murine CaCC increased with voltage and electrode Ca(2+) concentration. The current saturated at [Ca](CYT) of ∼1,000 nM and exhibited an EC(50) for Ca(2+) of ∼500 nM, independent of depolarization potential. Activation time constants were between 100 and 200 ms, independent of electrode Ca(2+). Repolarization-related tail currents elicited by stepping from +100 mV to varying test potentials exhibited deactivation time constants of 50-200 ms that increased with voltage when electrode [Ca](CYT) was 1,000 nM. The calmodulin inhibitor N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride (W-7, 30 μM) blocked CaCC. The myosin light chain kinase blockers 1-(5-iodonaphthalene-1-sulfonyl)-1H-hexahydro-1,4-diazepine hydrochloride (ML-7, 1-50 μM) and 1-(5-chloronaphthalene-1-sulfonyl)-1H-hexahydro-1,4-diazepine hydrochloride (ML-9, 10 μM) were similarly effective. Resting pericytes were hyperpolarized by ML-7. Pericytes exposed to ANG II (10 nM) depolarized from a baseline of -50 ± 6 to -29 ± 3 mV and were repolarized to -63 ± 7 mV by exposure to 50 μM ML-7. The Ca(2+)/calmodulin-dependent kinase inhibitor KN-93 reduced pericyte CaCC only when it was present in the electrode and extracellular buffer from the time of membrane break-in. We conclude that murine DVR pericytes are modulated by [Ca](CYT), membrane potential, and phosphorylation events, suggesting that Ca(2+)-dependent Cl(-) conductance may be a target for regulation of vasoactivity and medullary blood flow in vivo.
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