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Originally published In Press as doi:10.1074/jbc.M105345200 on December 21, 2001

J. Biol. Chem., Vol. 277, Issue 10, 8449-8456, March 8, 2002
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Identification and Localization of T-type Voltage-operated Calcium Channel Subunits in Human Male Germ Cells
EXPRESSION OF MULTIPLE ISOFORMS*

Suchitra JagannathanDagger , Emma L. PuntDagger §, Yuchun GuDagger , Christophe Arnoult||, Denny Sakkas**Dagger Dagger , Christopher L. R. Barratt**, and Stephen J. PublicoverDagger

From the Dagger  School of Biosciences, University of Birmingham, Birmingham B15 2TT, United Kingdom, || CNRS, Commissariat à l'Energie Atomique/Grenoble DBMS-CIS, 17 Rue des Martyrs, Grenoble, 38054 France, and the ** Reproductive Biology and Genetics Research Group, Assisted Conception Unit, Birmingham Women's Hospital, Birmingham B15 2TG, United Kingdom

Low voltage activated, voltage-operated Ca2+ channels are expressed in rodent male germ cells and are believed to be pivotal in induction of the acrosome reaction in mouse spermatozoa. However, in humans, very little is known about expression of voltage-operated Ca2+ channels in male germ cells or their function. We have used reverse transcription-polymerase chain reaction, in situ hybridization, and patch clamp recording to investigate the expression of low voltage activated voltage-operated Ca2+ channels in human male germ cells. We report that full-length transcripts for both alpha 1G and alpha 1H low voltage activated channel subunits are expressed in human testis. Multiple isoforms of alpha 1G are present in the testis and at least two isoforms of alpha 1H, including a splice variant not previously described in the human. Transcripts for all the isoforms of both alpha 1G and alpha 1H were detected by reverse transcription-polymerase chain reaction on mRNA isolated from human spermatogenic cells. In situ hybridization for alpha 1G and alpha 1H localized transcripts both in germ cells and in other cell types in the testis. Within the seminiferous tubules, alpha 1H was detected primarily in germ cells. Using the whole cell patch clamp technique, we detected T-type voltage-operated Ca2+ channel currents in isolated human male germ cells, although the current amplitude and frequency of occurrence were low in comparison to the occurrence of T-currents in murine male germ cells. We conclude that low voltage activated voltage-operated Ca2+ channels are expressed in cells of the human male germ line.

* This work was supported in part by a National Health Service Locally Organized Research Scheme project grant.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AJ420779.

§ Recipient of a Biotechnology and Biological Sciences Research Council studentship.

Present address: Medical Research Council Clinical Sciences Center, Division of Medicine, Hammersmith Campus, Imperial College School of Medicine, London W12 ONN, United Kingdom

Dagger Dagger Present address: Dept. of Obstetrics and Gynecology, Yale University School of Medicine, New Haven, CT 06520-8063.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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