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Apoptosis, Onset and Maintenance of Spermatogenesis: Evidence for the Involvement of Kit in Kit-Haplodeficient Mice¹

^a Physiopathologie de la Reproduction, Unité Mixte de Recherche 6073 "Physiologie de la Reproduction et des Comportements," Institut National de Recherche Agronomique, Centre National de la Recherche Scientifique, Université de Tours, Nouzilly, France ^b UMR 955 Institut National de Recherche Agronomique, Ecole Nationale Veterinaire d'Alfort, 94704 Maisons-Alfort, France

Kit/stem cell factor (SCF ) has been reported to be involved in survival and proliferation of male differentiating spermatogonial cells. This kinetics study was designed to assess the role of Kit/SCF during spermatogenesis in mice, and the extent of male programmed germ cell death was measured between 8 and 150 days of age. For this purpose, 129/Sv inbred mice in which one Kit allele was inactivated by an nlslacZ sequence insertion (Kit^W-lacZ/+) were compared with 129/Sv inbred mice with wild-type alleles at the Kit locus. Four different approaches were used: 1) morphometric study to assess spermatogenesis, 2) flow cytometry to study testicular cell ploidy, 3) in situ end labeling to detect apoptosis, and 4) follow-up of reporter gene expression. Spermatogenesis was lower in Kit^W-lacZ/+ heterozygous mice both at the onset of spermatogenesis and during adulthood. Indeed, greater apoptosis occurred at the onset of spermatogenesis. This was followed in the adult by a smaller seminiferous tubule diameter and a lower ratio between type B spermatogonia and type A stem spermatogonia in Kit^W-lacZ/+ mice compared with Kit^+/+ mice. These differences are probably related to the Kit haplodeficiency, which was the only difference between the two genotypes. Germ cell counts and testicular cell ploidy revealed delayed meiosis in Kit^W-lacZ/+ mice. Reporter gene expression confirmed expression of the Kit gene at the spermatogonial stage and also revealed Kit expression during the late pachytene/diplotene transition. These results suggest involvement of Kit/SCF at different stages of spermatogenesis.

First decision: 24 October 2001.

¹ This work was supported by the French Ministry of Education and Scientific Research, the National Institute for Agricultural Research, and the National Centre for Scientific Research as part of UMR 6073. V.C. held a grant from Organon Research Foundation (FARO, France).

² Correspondence: D. Royere, Biologie de la Reproduction, Centre Hospitalier Universitaire Bretonneau, 37044 Tours Cedex, France. FAX: 33 02 47 47 84 99; royere{at}med.univ-tours.fr

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