Beyond the Identification of Transcribed Sequences:
Functional, Evolutionary and Expression Analysis
12th International Workshop
October 25-28, 2002
Washington, DC

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RNAs Isolated from Ejaculate Spermatozoa Provide a Noninvasive Means to Investigate Testicular Gene Expression

G. Charles Ostermeier1, Robert J. Goodrich1, Kary Thompson2, John Rocket2, Michael P. Diamond1, Karen Collins1, NICHD Reproductive Medicine Network1, David J. Dix2, 3David Miller and Stephen A. Krawetz1,4,5
1Department of Obstetrics and Gynecology, 4Center for Molecular Medicine and Genetics, 5Institute for Scientific Computing, Wayne State University, Detroit, MI, USA.  2Reproductive Toxicology Division, National Health and Environmental Effects Research Laboratory, Environmental Protection Agency, Research Triangle Park, NC, USA, 3University of Leeds, U.K.
Telephone: 313-577-6770
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Idiopathic male infertility will affect at least 6% of all couples.   This is often not diagnosed until several IVF cycles have been completed.  This places couples at unnecessary risk.  The recent demonstration that mature ejaculate spermatozoa contain RNAs, may provide a window from which to view the gene expression history of spermatogenesis.  If true, ejaculate spermatozoa could be used as non-invasive surrogates for testis infertility investigations.  To assess whether spermatozoal transcripts encapsulate spermatogenic gene expression, a set of 27,016 unique expressed sequence tags (ESTs) was interrogated using cDNAs from a pool of testes representing 19 trauma victims and cDNAs from a pool of 9 individuals’ ejaculate spermatozoal mRNAs.  The testes cDNA hybridized to 7,157 ESTs and contained all 3,281 ESTs identified by the pooled-ejaculate probe.  The veracity of this observation was assessed by interrogating the same set of ESTs with cDNAs derived from the spermatozoal RNA of a single individual’s ejaculate.  All 2,784 of the ESTs identified were contained within the pooled testes cDNA and 2,780 of these were found within the pooled ejaculate probe.  These findings support the view that a spermatozoal mRNA fingerprint can encapsulate the gene expression of spermatogenesis.  The data further show that spermatozoal transcript populations vary among men.  Prior to spermatozoal RNA based diagnosis of infertility,the range of ‘normal’ must be established.  We have begun to address this issue by comparing spermatozoal transcript profiles among normal fertile men.  This analysis has revealed an invariant universal core and range of gene transcripts within ejaculate spermatozoa among fertile men.  We expect that those RNAs which are invariant represent a core set of messages necessary for the production of normal fertile spermatozoa.  Acknowledgments: This work was funded in part by NIH grant HD36512 and WSU grant 95200 to SAK and NIH grant HD39005 to MPD. GCO is supported in part by a Wayne State University's School of Medicine Dean's Post-Doctoral Fellowship award and the NICHD CIR-LRP.  The information in this document has been funded in part by the U.S. Environmental Protection Agency.  It has been subjected to review by the National Health and Environmental Effects Research Laboratory and approved for publication.  Approval does not signify that the contents reflect the views of the Agency, nor does mention of trade names or commercial products constitute endorsement or recommendation for use.

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