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Immunolocalization of protein kinase C isoenzymes , I, II, , and in mouse kidney

¹Institute of Pharmacology and Toxicology, University of Tübingen, 72074 Tübingen; ²Max-Planck-Institute for Experimental Endocrinology, 30625 Hannover, Germany; and ³Departments of Medicine and Pharmacology, University of California, and Veterans Affairs Medical Center, San Diego, California 92161

Submitted 7 August 2003 ; accepted in final form 22 March 2004

Localization of protein kinase C (PKC) isoenzymes , I, II, , and was studied employing Western blot analysis and immunohistochemical methods including confocal laser-scanning microscopy in the kidney of two mice strains, namely, C57BL/6 and 129/Sv, which have recently been used as genetic backgrounds for respective knockout mice. Immunoblot analysis identified immunoreactive bands for each isoenzyme in total kidney cell extracts. Isoenzyme expression sites were identical for both strains. Glomeruli expressed PKC-, -I, and -. The latter isoenzme was also detected in apical aspects of proximal convoluted but not in proximal straight tubules. In contrast to rats, neither PKC- nor PKC-I was detectable in the proximal tubule. Immunofluorescence was observed in luminal membranes of medullary (MTAL) and cortical thick ascending limbs for PKC-I and in MTAL for PKC-. The cortical collecting duct expressed PKC-, -I, and - in intercalated cells only. In the outer medullary collecting duct, PKC- and -I were detectable in principal cells, whereas PKC- was found in intercalated cells. In the inner medullary collecting duct, PKC-, -I, and -II were detected. As described for the rat, the expression of PKC-II was otherwise restricted to cortical and medullary interstitial cells. The specificity of all labeling was confirmed in respective PKC isoenzyme knockout mice. In summary, distinct expression patterns were shown for PKC isoenzymes , I, II, , and in the mouse kidney.

immunohistochemistry; knockout; renal interstitial cells

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