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Analysis of Cell-Type-Specific Gene Expression During Mouse Spermatogenesis¹

University Department of Growth and Reproduction,³ Rigshospitalet, Blegdamsvej 9, DK-2100 Copenhagen, Denmark Institute of Experimental Animals,⁴ Department of Pharmacology, St. Marianna University School of Medicine, Kawasaki, Japan The CREST Programme,⁵ The Japanese Science and Technology Agency, Tokyo, Japan

In rodents, changes in gene expression during spermatogenesis can be monitored by sampling testis from each day during postnatal development. However, changes in gene expression at the tissue level can reflect changes in the concentration of an mRNA in a specific cell type, changes in volume of specific cells, or changes in the cell-type composition. This reflects the cellularity of the tissue. Here we have combined techniques that assess the expression profiles of genes at the whole-tissue level, differential display and DNA array, and, at the level of cellularity, in situ hybridization. Combining results from these techniques allows determination of the cell-type-specific gene-expression patterns of many genes during spermatogenesis. Differential display was used to determine expression profiles with high sensitivity and independent of prior knowledge of the sequence, whereas DNA arrays quickly assess the expression profiles of all the genes. This identified three groups of gene-expression profiles. The major group corresponds to genes that are upregulated in spermatocytes during either the mid- or late- pachytene phase of spermatogenesis (stages VII–XI). This pachytene cluster was gradually extinguished in the later spermatid stages but was followed by another cluster of genes expressed in spermatids. Finally, a group of genes was downregulated during spermatogenesis and probably expressed in nongerm cells. We believe that expression of most genes can be described by a combination of these cell-type-specific expression patterns.

¹ Supported by The European Commission, The Japanese Science and Technology Agency CREST program, and Svend Andersen's foundation. The authors are solely responsible for statements made and they do not represent the opinion of the European Commission, which is not responsible for any use that might be made of data appearing herein.

² Correspondence: Kristian Almstrup, University Department of Growth and Reproduction, Section GR-5064, Rigshospitalet, Blegdamsvej 9, DK- 2100 Copenhagen, Denmark. FAX: 45 3545 6054;Kristian.Almstrup{at}biobase.dk

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