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Division of Nephrology, University of Maryland School of Medicine, Baltimore, Maryland 21201-1595
We devised a method for removal of pericytes from isolated descending vasa recta (DVR). After enzymatic digestion, aspiration of a descending vas rectum into a micropipette strips the pericytes from the abluminal surface. Pericytes and denuded endothelia can be recovered for separate study. Using fura 2-loaded preparations, we demonstrated that 10 nM angiotensin II (ANG II) elevates pericyte intracellular Ca2+ concentration ([Ca2+]i) and suppresses endothelial [Ca2+]i. The anion transport blocker probenecid helps retain fura 2 in the pericyte cytoplasm. DVR endothelia were accessed for membrane potential measurement by perforated-patch whole cell recording by using the pericyte-stripping technique and by turning nondigested vessels inside out with concentric micropipettes. By either method of access, 10 nM ANG II depolarized (n = 20) and 100 nM bradykinin hyperpolarized (n = 25) the endothelia. We conclude that isolated endothelia and pericytes remain functional for study of [Ca2+]i responses and that ANG II and bradykinin receptors exist separately on each cell type.
medulla; kidney; microcirculation; patch clamp; fura 2
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