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Identification of Markers for Precursor and Leydig Cell Differentiation in the Adult Rat Testis Following Ethane Dimethyl Sulphonate Administration¹

^a Department of Cell Biology and Histology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands ^b Banting and Best Department of Medical Research, University of Toronto, Toronto, Ontario, Canada M5G 1L6 ^c Institute for Hormone and Fertility Research, University of Hamburg, 22529 Hamburg, Germany

Administration of ethane dimethane sulphonate (EDS) to adult rats results in the destruction of all Leydig cells, followed by a complete regeneration. We investigated this regeneration process in more detail, using different markers for precursor and developing Leydig cells: the LH receptor, 3ß-hydroxysteroid dehydrogenase (3ß-HSD), transforming growth factor (TGF), and a new marker for Leydig cell maturation, relaxin-like factor (RLF).

LH receptor immunoreactivity was found in Leydig cell-depleted testes at 3 and 8 days after EDS administration. The positive (precursor) cells had a mesenchymal-like morphology. The number of LH receptor-positive cells 8 days after EDS administration was 15 ± 4 per 500 Sertoli cell nuclei. Fifteen days after EDS administration, the first new Leydig cells could be observed. These cells stained positively with both the antibodies against the LH receptor and 3ß-HSD, while some cells also stained positively for TGF.

After EDS administration, RLF mRNA disappeared from the testis and reappeared again at the time of the appearance of the first Leydig cells. Concomitant with the increase in the number of Leydig cells, the number of RLF-expressing cells increased.

The observations of the present study give further support to the hypothesis that Leydig cell development in the prepubertal testis, and in the adult testis following EDS administration, takes place along the same cell lineage and suggest, therefore, that the adult EDS-treated rat can serve as a model for studying the adult-type Leydig cell development that normally occurs in the prepubertal rat testis.

¹ K.J.T. is a recipient of a senior fellowship of the Royal Netherlands Academy of Arts and Sciences (KNAW fellow program). K.J.T. and J.H.D. are also supported by a NATO Collaborative Research Grant (no. CRG 940027). M.B. and R.I. are supported by the Deutsche Forschungsgemeinschaft (Grants Iv7/4–2–7 and Iv7/4–3–2).

² Correspondence: Katja J. Teerds, Department of Biochemistry, Cell Biology and Histology, Faculty of Veterinary Medicine, Utrecht University, P.O. Box 80.176, 3508 TD Utrecht, The Netherlands. FAX: 31 30 2516853; k.teerds{at}pobox.accu.uu.nl

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