Spermatogenic cell-somatic cell interactions are required for maintenance of spermatogenic cell glutathione.
Sertoli cells play a major role in the regulation of spermatogenic cell energy metabolism and differentiation. This study demonstrates that Sertoli cells are essential for the maintenance of spermatogenic cell glutathione (GSH), an important intracellular reductant and detoxicant. Primary spermatocytes and round spermatids isolated from Xenopus laevis contained 1.5 +/- 0.1 mM GSH, but sperm lacked detectable GSH. During a 5-day culture period, isolated spermatocytes and spermatids lost 80% of the initial GSH (t 1/2 = 55 h). The levels of GSH were unaffected by L-buthionine-SR-sulfoximine (BSO), a selective inhibitor of GSH synthesis. Cultures of testicular lobules and spermatocysts (composed of germ cells and Sertoli cells) depleted of interstitial tissue lost only 30% of their initial GSH in 4.5 days; the GSH levels decreased during treatment with BSO. Spermatogenic cells in cultured testes maintained their GSH levels for 7 days by a BSO-sensitive mechanism. These results demonstrate that the intracellular GSH levels of spermatogenic cells are dependent upon germ cell-somatic cell interactions. Spermatogenic cells were shown to possess gamma-glutamyl transpeptidase, glutathione synthetase, 5-oxoprolinase, and gamma-glutamylcysteine synthetase activities. [35S] Cysteine incorporation and distribution as analyzed by high performance liquid chromatography (HPLC) showed that isolated spermatogenic cells are capable of GSH synthesis. The rate of GSH synthesis, however, was insufficient to compensate for GSH turnover. These results demonstrate that production of spermatogenic cell GSH is dependent upon Sertoli cells. To our knowledge, this is the first evidence that interactions between different cell types may be of significance in GSH metabolism.
Li
L Y
LY
Department of Biochemistry, Cornell University Medical College, New York, New York 10021.
Seddon
A P
AP
Meister
A
A
Risley
M S
MS
D000818:Animals; D000963:Antimetabolites / Q000494:pharmacology; D019328:Buthionine Sulfoximine; D002450:Cell Communication; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D046508:Culture Techniques; D005978:Glutathione / Q000096:biosynthesis / Q000378:metabolism*; D008297:Male; D008717:Methionine Sulfoximine / Q000031:analogs & derivatives / Q000494:pharmacology; D012708:Sertoli Cells / Q000502:physiology*; D013087:Spermatids / Q000187:drug effects / Q000378:metabolism; D013090:Spermatocytes / Q000187:drug effects / Q000378:metabolism; D013091:Spermatogenesis; D013094:Spermatozoa / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism* / Q000648:ultrastructure; D013737:Testis / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D014982:Xenopus laevis
D000963:Antimetabolites; D008717:Methionine Sulfoximine; D019328:Buthionine Sulfoximine; D005978:Glutathione