Age affects gene expression in mouse spermatogonial stem-progenitor cells.pdfVIP

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Age affects gene expression in mouse spermatogonial stem-progenitor cells.pdf

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Age affects gene expression in mouse spermatogonial stem-progenitor cells

Age affects gene expression in mouse spermatogonial stem/ progenitor cells Maria Kokkinaki1, Tin-Lap Lee2, Zuping He1, Jiji Jiang1, Nady Golestaneh1, Marie-Claude Hofmann3, Wai-Yee Chan1,2, and Martin Dym1 1Department of Biochemistry and Molecular Cellular Biology, Georgetown University Medical Center, 3900 Reservoir Road NW, Washington, District of Columbia 20057, USA 2National Institute of Child Health and Human Development, NIH, Bethesda, Maryland 20892, USA 3Department of Veterinary Biosciences, University of Illinois, Urbana, Illinois 61802, USA Abstract Spermatogenesis in man starts with spermatogonial stem cells (SSCs), and leads to the production of sperm in ～64 days, common to old and young men. Sperm from elderly men are functional and able to fertilize eggs and produce offspring, even though daily sperm production is more than 50% lower and damage to sperm DNA is significantly higher in older men than in those who are younger. Our hypothesis is that the SSC/spermatogonial progenitors themselves age. To test this hypothesis, we studied the gene expression profile of mouse SSC/progenitor cells at several ages using microarrays. After sequential enzyme dispersion, we purified the SSC/progenitors with immunomagnetic cell sorting using an antibody to GFRA1, a known SSC/progenitor cell marker. RNA was isolated and used for the in vitro synthesis of amplified and labeled cRNAs that were hybridized to the Affymetrix mouse genome microarrays. The experiments were repeated twice with different cell preparations, and statistically significant results are presented. Quantitative RT-PCR analysis was used to confirm the microarray results. Comparison of four age groups (6 days, 21 days, 60 days, and 8 months old) showed a number of genes that were expressed specifically in the older mice. Two of them (i.e. Icam1 and Selp) have also been shown to mark aging hematopoietic stem cells. On the other hand, the expression levels of the genes encoding the SSC markers Gfra1 an