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Journal of Histochemistry and Cytochemistry, Vol. 48, 807-820, June 2000, Copyright © 2000, The Histochemical Society, Inc.

ARTICLE

Immunodetection of {alpha}1E Voltage-gated Ca2+ Channel in Chromogranin-positive Muscle Cells of Rat Heart, and in Distal Tubules of Human Kidney

Marco Weiergräbera, Alexey Pereverzeva, Rolf Vajnaa, Margit Henrya, Martin Schramma, Wolfgang Nastainczykb, Heike Grabschc, and Toni Schneidera
a Institute of Neurophysiology, University of Köln, Köln, Germany
b Institute of Medical Biochemistry and Molecular Biology, University of Saarland, Homburg/Saar, Germany
c Institute of Pathology, Heinrich-Heine-University of Düsseldorf, Düsseldorf, Germany

Correspondence to: Toni Schneider, University of Köln, Institute of Neurophysiology, Robert-Koch-Str. 39, D-50931 Köln, Germany. E-mail: Toni.Schneider@uni-koeln.de

The calcium channel {alpha}1E subunit was originally cloned from mammalian brain. A new splice variant was recently identified in rat islets of Langerhans and in human kidney by the polymerase chain reaction. The same isoform of {alpha}1E was detected in rat and guinea pig heart by amplifying indicative cDNA fragments and by immunostaining using peptide-specific antibodies. The apparent molecular size of cardiac {alpha}1E was determined by SDS-PAGE and immunoblotting (218 ± 6 kD; n = 3). Compared to {alpha}1E from stably transfected HEK-293 cells, this is smaller by 28 kD. The distribution of {alpha}1E in cardiac muscle cells of the conducting system and in the cardiomyoblast cell line H9c2 was compared to the distribution of chromogranin, a marker of neuroendocrine cells, and to the distribution of atrial natriuretic peptide (ANP). In serial sections from atrial and ventricular regions of rat heart, co-localization of {alpha}1E with ANP was detected in atrium and with chromogranin A/B in Purkinje fibers of the conducting system in both rat atrium and ventricle. The kidney is another organ in which natriuretic peptide hormones are secreted. The detection of {alpha}1E in the distal tubules of human kidney, where urodilatin is stored and secreted, led to the conclusion that the expression of {alpha}1E in rat heart and human kidney is linked to regions with endocrine functions and therefore is involved in the Ca2+-dependent secretion of peptide hormones such as ANP and urodilatin. (J Histochem Cytochem 48:807–819, 2000)

Key Words: {alpha}1E calcium channels, R-type Ca2+ channel, endocrine, chromogranin, atrial natriuretic peptide, urodilatin, H9c2 cells, conducting system, blood pressure regulation


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