title: Soluble Fas ligand activates the sphingomyelin pathway and induces apoptosis in luteal steroidogenic cells independently of stress-activated p38(MAPK).
abstract: Fas ligand (FasL) is implicated as a mediator of luteolysis. However, a gap exists in our understanding of the Fas-mediated signaling mechanisms that are involved in either the loss of progesterone production or the structural regression of the corpus luteum. In the present study we investigated the acute and chronic effects of FasL with respect to activation of cytokine/stress-induced signaling pathways and apoptosis in bovine steroidogenic cells. More specifically, we investigated soluble FasL (sFasL)-activated production of ceramide, a second messenger of the sphingomyelin pathway, and activation of p38(MAPK), a member of the MAPK family. sFasL activated the sphingomyelin pathway, as evidenced by a 2-fold increase (P < 0.05) in the production of ceramide. Pretreatment with imipramine (50 micro M), an inhibitor of acid sphingomyelinase activity, attenuated (75%; P < 0.05) sFasL-induced ceramide production, suggesting that the increase in ceramide was partially the result of acid sphingomyelinase-mediated hydrolysis of sphingomyelin. Treatment of luteal cells with sFasL or a cell-permeable ceramide analog (C8) for 24-48 h resulted in a significant increase (P < 0.05) in apoptosis. Western blot analysis revealed that sFasL had little effect on the activation of p38(MAPK) in primary bovine luteal steroidogenic cells. Furthermore, pretreatment with the p38(MAPK) inhibitor SB203580 failed (P > 0.05) to inhibit sFasL- or C8-induced death. Although sFasL did not alter basal progesterone levels detected in the culture medium, C8 caused a significant increase (P < 0.05) in progesterone concentrations within the medium. Collectively, these data suggest that the role of FasL in luteolysis may be to activate the stress-induced sphingomyelin pathway that, in turn, serves as a mediator of apoptosis.
authors.affiliation: Vincent Center for Reproductive Biology, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.
authors.forename: James K
authors.initials: JK
authors.lastname: Pru
authors.forename: Isabel R
authors.initials: IR
authors.lastname: Hendry
authors.forename: John S
authors.initials: JS
authors.lastname: Davis
authors.forename: Bo R
authors.initials: BR
authors.lastname: Rueda
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D002417:Cattle; D002518:Ceramides / Q000494:pharmacology; D003338:Corpus Luteum / Q000166:cytology / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D053222:Fas Ligand Protein; D005260:Female; D006868:Hydrolysis; D008562:Membrane Glycoproteins / Q000494:pharmacology*; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D011247:Pregnancy; D011374:Progesterone / Q000096:biosynthesis; D013108:Sphingomyelin Phosphodiesterase / Q000037:antagonists & inhibitors / Q000378:metabolism; D013109:Sphingomyelins / Q000378:metabolism*; D013256:Steroids / Q000096:biosynthesis*; D013312:Stress, Physiological; D048051:p38 Mitogen-Activated Protein Kinases
chemical_list: D002518:Ceramides; D004791:Enzyme Inhibitors; D053222:Fas Ligand Protein; D008562:Membrane Glycoproteins; D013109:Sphingomyelins; D013256:Steroids; D011374:Progesterone; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; D013108:Sphingomyelin Phosphodiesterase