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BOR - Papers in Press, published online ahead of print February 11, 2004.
Biol Reprod 2004, 10.1095/biolreprod.103.026575
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Submitted December 15, 2003
Returned for revision December 30, 2003
Accepted February 3, 2004

Testis


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

Kristian Almstrup *, John E. Nielsen , Martin A. Hansen , Masami Tanaka , Niels E. Skakkebaek , and Henrik Leffers

* To whom correspondence should be addressed. E-mail: kristian.almstrup{at}biobase.dk.

Abstract
In rodents, changes in gene expression during spermatogenesis can be monitored by sampling testis from each day during post natal development. However, changes in gene expression at the tissue level can reflect changes in the concentration 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 up-regulated in spermatocytes during either the mid or late pachytene phase of spermatogenesis (stage 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 down-regulated during spermatogenesis and probably expressed in non-germ cells. We believe that expression of most genes can be described by a combination of these cell type specific expression patterns.

Key words: Testis • Developmental biology • Gene regulation • Spermatogenesis


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