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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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Submitted August 1, 2003
Returned for revision August 28, 2003
Accepted November 10, 2003

Testis


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

Marjut Otala , Laura Suomalainen *, Markku O. Pentikäinen , Petri Kovanen , Mikko Tenhunen , Krista Erkkilä , Jorma Toppari , and Leo Dunkel

* To whom correspondence should be addressed. E-mail: laura.suomalainen{at}hus.fi.

Abstract
Male germ cells are susceptible to radiation-induced injury and infertility is a common problem after total body irradiation. Here we investigated, firstly, the effects of irradiation on germ cells in mouse testis and, secondly, 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 hours 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 hours 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.

Key words: Male Reproductive Tract • Testis • Apoptosis • Spermatogenesis • Stress


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