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BOR - Papers in Press, published online ahead of print February 16, 2005.
Biol Reprod 2005, 10.1095/biolreprod.104.037499
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Submitted October 28, 2004
Returned for revision December 6, 2004
Accepted February 10, 2005

Testis


Gene Expression in Rat Leydig Cells During Development from the Progenitor to Adult Stage: A Cluster Analysis

Ren-Shan Ge , Qiang Dong , Chantal M. Sottas , Haolin Chen , Barry R. Zirkin , and Matthew P. Hardy *

* To whom correspondence should be addressed. E-mail: hardy{at}popcbr.rockefeller.edu.

Abstract
The postnatal development of Leydig cells can be divided into three distinct stages: initially they exist as fibroblast-like progenitor Leydig cells (PLCs) appearing in the testis by days 14 to 21; subsequently, by day 35, they become immature Leydig cells (ILCs) acquiring steroidogenic organelle structure and enzyme activities but metabolizing most of the testosterone they produce; finally, as adult Leydig cells (ALCs) by day 90 they actively produce testosterone. The factors controlling proliferation and differentiation of Leydig cells remain largely unknown, and the aim of the present study was to identify changes in gene expression during development through cDNA array analysis of PLCs, ILCs and ALCs. By cluster analysis, it was determined that the transitions from PLC to ILC to ALC were associated with downregulation of mRNAs corresponding to 107 genes. The down-regulated genes included cell cycle regulators e.g. cyclin D1 (Ccnd1), growth factors e.g. basic fibroblast growth factor (Fgf2), growth factor related receptors e.g. platelet-derived growth factor {alpha} receptor (Pdgfra), oncogenes e.g. kit oncogene (Kit) and transcription factors e.g. early growth response 1 (Egr1). Conversely, expression levels of 264 genes were increased by at least 2-fold. Most of these were related to differentiated function and included steroidogenic enzymes e.g. 11{beta}-hydroxysteroid dehydrogenase 2 (Hsd11b2), neurotransmitter receptors e.g. acetylcholine receptor nicotinic a 4 (Chrna4), stress response factors e.g. glutathione transferase 8 (Gsta4), and protein turnover enzymes e.g. tissue inhibitor of metalloproteinase 2 (Timp2). The detection of Hsd11b2 mRNA in the array was the first indication that this gene is expressed in Leydig cells, and parallel increases in Hsd11b2 mRNA and enzyme activity were recorded. Thus, gene profiling demonstrates that postnatal development is associated with changes in the expression levels of several different clusters of genes consistent with the processes of Leydig cell growth and differentiation.

Key words: Developmental biology • Gene regulation • Leydig cells • Luteinizing hormone • Steroid hormone receptors


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