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MOLECULAR ANALYSIS OF HUMAN SPERMATOZOA: POTENTIAL FOR INFERTILITY RESEARCH AND SCREENING
Citation:
Miller, D., D J. Dix, R. Reid, S. Wykes, AND S. A. Krawetz. MOLECULAR ANALYSIS OF HUMAN SPERMATOZOA: POTENTIAL FOR INFERTILITY RESEARCH AND SCREENING. Presented at Gender Differences in Reproductive Biology and Toxicology, Tucson, AZ, November 09 - 11, 2000.
Description:
Molecular Analysis of Human Spermatozoa: Potential for Infertility Research and Screening
David Miller1, David Dix2, Robert Reid3, Susan Wykes3 and Stephen Krawetz3
1Reproductive Biology Group, University of Leeds, UK
2Reproductive Toxicology Division, U.S. Environmental Protection Agency, USA
3Department of Ob/Gyn, CMMG & ICS, Wayne State University, USA
Approximately 1 in 6 couples will experience difficulty in conceiving a child. This can be traced to male-factor infertility in at least half of these cases. Although, chromosomal deletions like Y and several single-gene defects have been observed, it is astonishing that the cause of over 75% of the cases of non-obstructive azoospermia and oligozoospermia remain largely unknown. Using RT-PCR, in situ hybridization, cDNA library sequencing and cDNA microarrays, we have been able to demonstrate that both human and murine ejaculate spermatozoa contain a complex repertoire of mRNAs. Examples have included ?-actin, HSP 70, HSP 90 that are common to most cell types like and those specific to the spermatozoon like protamine 1, protamine 2 and transition protein 2. In addition, a series of unique new transcripts have also been identified using the cDNA sequencing and microarray screening strategies. While the identity and function of these mRNAs is unknown, we believe that they can provide a unique window into the events that occur during spermatogenesis. They may prove highly informative in the search for genes associated with male factor infertility. At present, we are utilizing a set of cDNA arrays to survey
the distribution of over 30,000 expressed sequence tags from testis RNAs and the RNAs from ejaculate spermatozoa of normal fertile men. This will define the NORMAL FERTILE MALE transcript fingerprint. Establishing this baseline will then permit us to define the corresponding spermatozoa RNA fingerprint(s) that are characteristic of the idiopathic infertile male. This data should enable us to identify new genes linked to male infertility. Application of this array
technology to monitor changes in expression profiles that may indicate perturbations in testicular gene expression associated with deleterious exposure to environmental factors including exposure to xenoestrogens is also being pursued.
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