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Articles in PresS, published online ahead of print April 23, 2002
Am J Physiol Renal Physiol, 10.1152/ajprenal.00065.2002
Submitted on February 14, 2002
Accepted on April 16, 2002
1 Department of Medicine, Division of Nephrology, University of Maryland School of Medicine, Baltimore, MD, USA
* To whom correspondence should be addressed. E-mail: tpallone{at}medicine.umaryland.edu.
We devised a method for removal of the pericytes from isolated descending vasa recta (DVR). After enzymatic digestion, aspiration of a DVR into a micropipette strips the pericytes from the abluminal surface. Both pericytes and denuded endothelia can be recovered for separate study. Using fura2 loaded preparations, we demonstrated that angiotensin II (AngII, 10 nM) elevates pericyte intracellular Ca2+ ( [Ca2+]i ) and suppresses endothelial [Ca2+]i. The anion transport blocker, probenecid, helps retain fura2 in the pericyte cytoplasm. DVR endothelia were accessed for membrane potential measurement by perforated patch whole cell recording using the pericyte stripping technique and by turning nondigested vessels inside out with concentric micropipettes. By either method of access, AngII (10 nM) depolarized (n = 20) and bradykinin (BK, 100 nM) hyperpolarized (n = 25) the endothelia. We conclude that isolated endothelia and pericytes remain functional for study of [Ca2+]i responses and that AngII and BK receptors exist separately on each cell type.
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