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1 Institute of Pharmacology and Toxicology, University of Tuebingen, Tuebingen, Germany
2 Max-Planck-Institute for Experimental Endocrinology, Hannover, Germany
3 Institute of Pharmacology and Toxicology, University of Tuebingen, Tuebingen, Germany; Departments of Medicine and Pharmacology, University of California San Diego, San Diego, CA, USA
* To whom correspondence should be addressed. E-mail: vvallon{at}ucsd.edu.
Localization of protein kinase C (PKC) isoenzymes alpha, beta I, beta II, delta, and epsilon 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 alpha, beta I and epsilon. The latter isoenzme was also detected in apical aspects of proximal convoluted but not in proximal straight tubules. In contrast to rat, neither PKC alpha nor PKC beta I were detectable in proximal tubule. Immunofluorescence was observed in luminal membranes of medullary and cortical thick ascending limbs (MTAL, CTAL) for PKC beta I and in MTAL for PKC epsilon. Cortical collecting duct (CCD) expressed PKC alpha, beta I, and delta in intercalated cells only. In outer medullary collecting duct (OMCD), PKC alpha and beta I were detectable in principal cells whereas PKC delta was found in intercalated cells. In the inner medullary collecting duct (IMCD), PKC alpha, beta I and beta II were detected. As described for the rat, the expression of PKC beta 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 alpha, beta I, beta II, delta and epsilon in mouse kidney.
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