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Testes Exhibit Elevated Expression of Calcitonin Gene-Related Peptide Receptor Component Protein¹

Geriatric Research, Education, and Clinical Center, Veterans Affairs Medical Center, and the Departments of Medicine (W.B., E.L.O., G.A.H., B.A.R.), Biochemistry and Molecular Biology (G.A.H.), and Neurology (B.A.R.), University of Miami School of Medicine, Miami, Florida 33101

Address all correspondence and requests for reprints to: Wayne Balkan, Ph.D., Veterans Affairs Medical Center, 1201 NW 16th Street, Geriatric Research, Education, and Clinical Center (11GRC), Miami, Florida 33125. E-mail: wbalkan{at}mednet.med.miami.edu

Calcitonin gene-related peptide (CGRP) receptor component protein (RCP) is a novel protein that modulates CGRP responsiveness in a variety of cell types. Using probes based on the isolation of CGRP-RCP complementary DNA (cDNA) from a guinea pig organ of Corti cDNA library, we cloned human (h) and mouse (m) CGRP-RCP cDNAs, both of which encode 148-residue proteins that at the amino acid levels are approximately 88% identical to each other and to the 146-residue guinea pig CGRP-RCP. Northern blot analysis confirmed the presence of CGRP-RCP messenger RNA in all of the human and mouse tissues tested. In these human tissues, hCGRP-RCP messenger RNA (major band at 3.1 kb, minor band at 7.5 kb) was most prevalent in the testis. In the mouse, the highest abundance of CGRP-RCP RNA was clearly in the testis (major band at 1.6 kb, minor band at 1.1 kb). Based on this tissue distribution of RNA, we sought to identify the cells in the murine testis that contained CGRP-RCP protein. Numerous antisera generated against hCGRP-RCP, including one to recombinant hCGRP-RCP, exhibited strong immunoreactivity localized to the head region of spermatozoa. No CGRP-RCP immunoreactivity was observed in other cells at less mature stages of sperm maturation, in Sertoli or interstitial (Leydig) cells, or in human spermatozoa. Murine epididymal (mature) spermatozoa exhibited CGRP-RCP immunoreactivity identical to that of testicular spermatozoa. Spermatozoa that underwent an experimentally induced acrosome reaction (acrosomal discharge) lost their CGRP-RCP immunoreactivity. Therefore, it appears that CGRP-RCP is associated with the acrosome, suggesting that it may play an important role in reproduction.

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