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Physiol. Genomics (November 15, 2005). doi:10.1152/physiolgenomics.00215.2004
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Submitted on September 16, 2004
Accepted on November 13, 2005

Expression Profiling of Purified Male Germ Cells: Stage Specific Expression Patterns Related to Meiosis and Post-Meiotic Development

Alan L. Y. Pang1*, Warren Johnson1, Neelakanta Ravindranath1, Martin Dym2, Owen M Rennert1, and Wai-Yee Chan3

1 NICHD/LCG/SDG, National Institutes of Health, Bethesda, MD, USA
2 Department of Cell Biology, Georgetown University, Washington, DC, USA
3 NICHD/LCG/SDG, National Institutes of Health, Bethesda, MD, USA; Department of Cell Biology, Georgetown University, Washington, DC, USA; Department of Pediatrics, Georgetown University, Washington, DC, USA

* To whom correspondence should be addressed. E-mail: panga{at}mail.nih.gov.

Gene expression profiling was performed using the NIA 15,000 cDNA microarray to reveal the differential expression pattern of 160 genes between meiotic pachytene spermatocytes and post-meiotic round spermatids of the mouse. Our results indicate that more genes are expressed in spermatids than in spermatocytes. Genes participating in cell cycle regulation and chromatin structure and dynamics are preferentially expressed in spermatocytes; while genes for protein turnover, signal transduction, energy metabolism and intracellular transport are prevalent in spermatids. This suggests a switch of functional requirement occurs when meiotic germ cells differentiate into haploid spermatids. Concordant expression patterns were obtained when quantitative real-time polymerase chain reaction was performed to verify the microarray data. Interestingly, the majority of the differentially expressed genes were under-represented in mitotic type A spermatogonia, and they were preferentially expressed in the testis. Our results suggest that an even higher proportion of the mouse genome is devoted to male gamete development from meiosis than was previously estimated. We also provide evidence that underscores the advantage of using purified germ cells over whole testes in profiling spermatogenic gene expression to identify transcripts that demonstrate stage-specific expression patterns.




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