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Journal of Histochemistry and Cytochemistry, Vol. 47, 981-994, August 1999, Copyright © 1999, The Histochemical Society, Inc.

ARTICLE

Immunohistochemical Detection of {alpha}1E Voltage-gated Ca2+ Channel Isoforms in Cerebellum, INS-1 Cells, and Neuroendocrine Cells of the Digestive System

Heike Grabschc, Alexei Pereverzeva, Marco Weiergräbera, Martin Schramma, Margit Henrya, Rolf Vajnaa, Ruth E. Beattied, Steve G. Volsend, Udo Klöcknerb, Jürgen Heschelera, and Toni Schneidera
a Institutes of Neurophysiology, University of Cologne, Cologne, Germany
b Vegetative Physiology, University of Cologne, Cologne, Germany
c Institute of Pathology, Klinikum Leverkusen, Leverkusen, Germany
d CNS Research, Ely Lilly & Company, Lilly Research Center, Windlesham, Surrey, United Kingdom

Correspondence to: Toni Schneider, Univ. of Cologne, Institute of Neurophysiology, Robert-Koch-Str. 39, D-50931 Köln, Germany. Fax 49-221-4786965.

Polyclonal antibodies were raised against a common and a specific epitope present only in longer {alpha}1E isoforms of voltage-gated Ca2+ channels, yielding an "anti-E-com" and an "anti-E-spec" serum, respectively. The specificity of both sera was established by immunocytochemistry and immunoblotting using stably transfected HEK-293 cells or membrane proteins derived from them. Cells from the insulinoma cell line INS-1, tissue sections from cerebellum, and representative regions of gastrointestinal tract were stained immunocytochemically. INS-1 cells expressed an {alpha}1E splice variant with a longer carboxy terminus, the so-called {alpha}1Ee isoform. Similarily, in rat cerebellum, which was used as a reference system, the anti-E-spec serum stained somata and dendrites of Purkinje cells. Only faint staining was seen throughout the cerebellar granule cell layer. After prolonged incubation times, neurons of the molecular layer were stained by anti-E-com, suggesting that a shorter {alpha}1E isoform is expressed at a lower protein density. In human gastrointestinal tract, endocrine cells of the antral mucosa (stomach), small and large intestine, and islets of Langerhans were stained by the anti-E-spec serum. In addition, staining by the anti-E-spec serum was observed in Paneth cells and in the smooth muscle cell layer of the lamina muscularis mucosae. We conclude that the longer {alpha}1Ee isoform is expressed in neuroendocrine cells of the digestive system and that, in pancreas, {alpha}1Ee expression is restricted to the neuroendocrine part, the islets of Langerhans. {alpha}1E therefore appears to be a common voltage-gated Ca2+ channel linked to neuroendocrine and related systems of the body. (J Histochem Cytochem 47:981–993, 1999)

Key Words: {alpha}1E calcium channels, cerebellum, INS-1 insulinoma cell line, secretion, endocrine digestive system, islets of Langerhans, chromogranin A, gastric antrum, small intestine, large intestine, Paneth cells, immunocytochemistry


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