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Regular Article |
a Department of Cell Biology, UMCU, 3584 CX Utrecht, The Netherlands
b Department of Biomedicine, Biomedicum, FIN-00014, University of Helsinki, Finland
c Department of Cellular and Molecular Biology and Pathology, University of Naples, 80131 Naples, Italy
With a novel method of eliminating spermatogenesis in host animals, male germ cells isolated from mice with targeted overexpression of glial cell line-derived neurotrophic factor (GDNF) were transplanted to evaluate their ability to reproduce the phenotype previously found in the transgenic animals. Successful depletion of endogenous spermatogenesis was achieved using fractionated ionizing irradiation. A dose of 1.5 Gy followed by a dose of 12 Gy after 24 h reduced the percentage of tubule cross-sections displaying endogenous spermatogenesis to approximately 3% and 10% as evidenced by histologic evaluation of testes at 12 and 21 wk, respectively, after irradiation. At this dose, no apparent harmful side effects were noted in the animals. Upon transplantation, GDNF-overexpressing germ cells were found to be able to repopulate the irradiated testes and to form clusters of spermatogonia-like cells resembling those found in the overexpressing donor mice. The cluster cells in transplanted host testes expressed human GDNF, as had been shown previously for clusters in donor animals, and both were strongly positive for the tyrosine kinase receptor Ret. Thus, we devised an efficient method for depleting the seminiferous epithelium of host mice without appreciable adverse effects. In these host mice, GDNF-overexpressing cells reproduced the aberrant phenotype found in the donor transgenic mice.
1 This work was supported by NIH grant HD 36476-02.
2 Correspondence: Laura B. Creemers, Department of Cell Biology, HP G02.525, UMCU, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands. FAX: 31 30 2541797; l.b.creemers{at}lab.azu.nl
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