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BOR - Papers in Press, published online ahead of print November 12, 2003.
Biol Reprod 2003, 10.1095/biolreprod.103.021840
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BIOLOGY OF REPRODUCTION 70, 759–767 (2004)
DOI: 10.1095/biolreprod.103.021840
© 2004 by the Society for the Study of Reproduction, Inc.


Testis

Protection from Radiation-Induced Male Germ Cell Loss by Sphingosine-1-Phosphate1

Marjut Otala3, Laura Suomalainen2,3, Markku O. Pentikäinen4, Petri Kovanen4, Mikko Tenhunen6, Krista Erkkilä3, Jorma Toppari5, and Leo Dunkel3

Programme for Developmental and Reproductive Biology,3 Biomedicum Helsinki and Hospital for Children and Adolescents, University of Helsinki, FIN-00029 HUS, Helsinki, Finland Wihuri Research Institute,4 FIN-00140, Helsinki, Finland Departments of Pediatrics and Physiology,5 University of Turku, FIN-20520, Turku, Finland Department of Oncology,6 Helsinki University Hospital, FIN-00029, Helsinki, Finland

Male germ cells are susceptible to radiation-induced injury, and infertility is a common problem after total-body irradiation. Here we investigated, first, the effects of irradiation on germ cells in mouse testis and, second, the role of sphingosine-1-phosphate (S1P) treatment in radiation-induced male germ cell loss. Irradiation of mouse testes mainly damaged the early developmental stages of spermatogonia. The damage was seen by means of DNA flow cytometry 21 days after irradiation as decreasing numbers of spermatocytes and spermatids with increasing amounts of ionizing radiation (0.1–2.0 Gy). Intratesticular injections of S1P given 1–2 h before irradiation (0.5 Gy) did not protect against short-term germ cell loss as measured by in situ end labeling of DNA fragmentation 16 h after irradiation. However, after 21 days, in the S1P-treated testes, the numbers of primary spermatocytes and spermatogonia at G2 (4C peak as measured by flow cytometry) were higher at all stages of spermatogenesis compared with vehicle-treated testes, indicating protection of early spermatogonia by S1P, whereas the spermatid (1C) populations were similar. In conclusion, S1P appears to protect partially (16%–47%) testicular germ cells against radiation-induced cell death. This warrants further studies aimed at development of therapeutic agents capable of blocking sphingomyelin-induced pathways of germ cell loss.

1 Supported by the Foundation for Pediatric Research (Finland), the Pediatric Graduate School (Finland), the Sigrid Jusélius Foundation (Finland), the Cancer Society of Finland, the Nona and Kullervo Väre Foundation (Finland), and Helsinki Biomedical Graduate School (Finland), Research and Science Foundation of Farmos (Finland). M.O. and L.S. contributed equally to this work.

2 Correspondence: Laura Suomalainen, Hospital for Children and Adolescents, Programme for Developmental and Reproductive Biology (5th Floor, Room B529b), Biomedicum Helsinki, University of Helsinki, P.O. Box 700, FIN-00029 HUS, Helsinki, Finland. FAX: 358 9 4717 1947; laura.suomalainen{at}hus.fi




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