Molecular Human Reproduction, Vol. 6, No. 4, 303-313,
April 2000
© 2000 European Society of Human Reproduction and Embryology
Testis and spermatogenesis |
Molecular characterization of a voltage-gated potassium channel expressed in rat testis*
1 Department of Obstetrics and Gynecology, 2 Department of Research, North Shore University Hospital–New York University School of Medicine, Manhasset, New York, 3 Department of Obstetrics and Gynecology and 4 Department of Cell Biology, New York University School of Medicine, New York, New York, USA
Abstract
Potassium (K+) channels are present in both mammalian testis and spermatozoa. Immunofluorescent detection of sperm-bound biotinylated charybdotoxin, an inhibitor of Ca2+-activated and of delayed rectifier K+ channels, indicated that these ion channels are uniformly distributed over the surface of both heads and tails of unfixed rat epididymal spermatozoa. Reverse transcription–polymerase chain reaction (RT–PCR) analysis on rat testis RNA with PCR primers, based on known nucleotide sequences of different classes of K+ channels, amplified sequences homologous to delayed rectifier K+ channels. In-situ RT–PCR on rat testis sections showed that these K+ channel transcripts are present in the cytoplasm of primary spermatocytes and post-meiotic elongating spermatids. Northern blot analysis of various rat tissues identified multiple K+ channel transcripts, some of which were observed only in testis. An attempt to obtain a full length rat testis K+ channel cDNA sequence gave an assembled sequence of 2693 base pairs with >90% homology to a delayed rectifier K+ channel, Kv1.3. A method for rapid amplification of cDNA ends was employed to amplify the 5' sequences of the rat testis cDNA but a unique sequence could not be obtained. DNA sequencer traces suggest that multiple related K+ channels which differed at their 5' ends were amplified in rat testis.
charybdotoxin labelling/potassium channel/sperm membrane/spermatozoa/testis
Notes
5 To whom correspondence should be addressed at: North Shore University Hospital, 300 Community Drive, Boas–Marks Biomedical Science Research Center – Room 119, Manhasset, New York 11030, USA
* Presented, in part, at the 53rd Annual Meeting of the American Society for Reproductive Medicine, Cincinnati, Ohio, October 18–22, 1997.
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