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GENE EXPRESSION IN THE TESTES OF NORMOSPERMIC VERSUS TERATOSPERMIC DOMESTIC CATS USING HUMAN CDNA MICROARRAY ANALYSES
Pukazhenthi, B. S., J C. Rockett, O. Ming, D J. Dix, P. Georgopoulos, W. J. Welsh, AND D. E. Wildt. GENE EXPRESSION IN THE TESTES OF NORMOSPERMIC VERSUS TERATOSPERMIC DOMESTIC CATS USING HUMAN CDNA MICROARRAY ANALYSES. Presented at Society for the Study of Reproduction, Cincinnati, Ohio, July 19-22, 2003.
GENE EXPRESSION IN THE TESTES OF NORMOSPERMIC VERSUS TERATOSPERMIC DOMESTIC CATS USING HUMAN cDNA MICROARRAY ANALYSES
B.S. Pukazhenthi1, J. C. Rockett2, M. Ouyang3, D.J. Dix2, J.G. Howard1, P. Georgopoulos4, W.J. J. Welsh3 and D. E. Wildt1
1Department of Reproductive Sciences, Smithsonian National Zoological Park, Washington, DC; 2Reproductive Toxicology Division, NHEERL, ORD, U.S. Environmental Protection Agency, Research Triangle Park, NC; 3Informatics Institute, University of Medicine and Dentistry of New Jersey, Piscataway, NJ; 4Environmental and Occupational Health Sciences Institute, Rutgers University at University of Medicine and Dentistry of New Jersey, Piscataway, NJ
Teratospermia (high incidence of abnormally shaped sperm/ejaculate) is prevalent in felids, including some domestic cats and several rare, wild felids. Species or populations lacking genetic variability produce more malformed sperm, but the molecular mechanisms of this phenomenon have not been established. This study analyzed testicular gene expression in normospermic (N; >60% normal sperm) versus teratospermic (T; <40% normal sperm) domestic cats. Total RNA was isolated from testes of N (n = 5) and T (n = 6) males using TrizolO reagent followed by DNase treatment and reverse transcription in the presence of [32P] dCTP. Membranes (Clontech Human Toxicology 1.2) were hybridized overnight (65?C), washed and exposed to autoradiographic film. Images were digitized by densitometer and analyzed with AtlasImage 2.0 software (Clontech). Data were normalized, transformed and differentially expressed genes identified using Bayesian analysis. Of the 1,185 genes represented on the membrane, 421 and 419 genes were expressed in both the N and T cats, respectively. Compared to the N controls, expression of 71 genes was up-regulated (P<0.05), and 97 genes were down-regulated (P<0.05) in T cats. Increased expression was observed in 6 DNA repair and recombination genes, 5 transcription genes, 4 cell cycle genes and 4 apoptosis genes. In contrast, decreased expression was observed in 2 DNA repair and recombination genes, 18 transcription genes, 6 cell cycle genes and 10 apoptosis genes. Data suggest that perturbation of multiple mechanisms (including DNA repair and recombination, transcription, cell cycle regulation and apoptosis during spermatogenesis) may give rise to teratospermia in the domestic cat. (Supported by NIH KO1 RR00135. This is an abstract of a proposed presentation and does not necessarily reflect EPA policy.)
KEY WORDS: cat, testis, teratospermia, microarray, gene expression