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USING DNA MICROARRAYS TO CHARACTERIZE GENE EXPRESSION IN TESTES OF FERTILE AND INFERTILE HUMANS AND MICE
Citation:
Rockett, J C., J. C. Luft, J B. Garges, M. S. Ricci, P. Patrizio, N. B. Hecht, AND D J. Dix. USING DNA MICROARRAYS TO CHARACTERIZE GENE EXPRESSION IN TESTES OF FERTILE AND INFERTILE HUMANS AND MICE. Presented at Functional Genomics and MicroArray Data Mining, Duke University, Durham, NC, August 3-4, 2000.
Description:
USING DNA MICROARRAYS TO CHARACTERIZE GENE EXPRESSION
IN TESTES OF FERTILE AND INFERTILE HUMANS AND MICE
John C. Rockett1, J. Christopher Luft1, J. Brian Garges1, M. Stacey Ricci2, Pasquale Patrizio2, Norman B. Hecht2 and David J. Dix1
Reproductive Toxicology Division, NHEERL, U.S. Environmental Protection Agency, RTP, NC1, and 2Center for Research on Reproduction and Women?s Health, University of Pennsylvania, Philadelphia, PA
Commercial filter-based DNA microarrays were used to assess gene expression in human and mouse testis. Two sources of human testis RNA were used: commercially available RNA from a pool of 19 human subjects, or RNA extracted from testicular biopsies of individual patients. Hybridizing cDNAs from the pooled human testis RNA to GeneFilter (Research Genetics) arrays containing over 20,000 known genes and ESTs led to the identification of approximately 800 known genes and thousands of ESTs with hybridization signals > 5-fold above background. Hybridization of Atlas (Clontech) human cDNA Expression and Stress/Toxicology filter arrays with cDNAs from pooled RNA and RNA from a testicular biopsy of a fertile patient confirmed results with the GeneFilters and identified additional genes expressed in human testis. 372 of the 822 human genes on these Atlas arrays yielded hybridization signals > 2-fold above background. RNA from three separate testicular biopsies from azoospermic patients were also reverse-transcribed and used to hybridize Atlas (Clontech) arrays. Each of these three unique clinical samples, diagnosed as either maturation arrest or Sertoli cell only, resulted in unique expression results for the 822 genes assessed by Atlas arrays. Genes expressed in the human testis are being compared to those expressed in the testes of wild-type mice (C57BL/6) and mutant mouse models of male infertility. Atlas mouse cDNA Expression and Stress/Toxicology filter arrays (Clontech) identified 581 genes expressed in wild-type mouse testis. More thorough analysis of these array results may reveal common patterns of gene expression in the testes of the two species, and between the various cases of infertility in humans and mice. (This is an abstract of a proposed presentation and does not necessarily reflect EPA policy.)