Phosphatidylinositol 3-kinase pathway regulates sperm viability but not capacitation on boar spermatozoa.
Phosphatidylinositol 3-kinase (PI3-K) plays an important role in cell survival in somatic cells and recent data pointed out a role for this kinase in sperm capacitation and acrosome reaction (AR). This study was undertaken to evaluate the role of PI3-K pathway on porcine spermatozoa capacitation, AR, and viability using two unrelated PI3-K inhibitors, LY294002 and wortmannin. In boar spermatozoa, we have identified the presence of PDK1, PKB/Akt, and PTEN, three of the main key components of the PI3-K pathway. Incubation of boar sperm in a capacitating medium (TCM) caused a significant increase in the percentage of capacitated (25 +/- 2 to 34 +/- 1% P < 0.05, n = 6) and acrosome reacted (1 +/- 1 to 11 +/- 1% P < 0.01, n = 6) spermatozoa compared with sperm in basal medium (TBM). Inhibition of PI3-K did affect neither the capacitation status nor AR nor protein p32 tyrosine phosphorylation of boar spermatozoa incubated in TBM or TCM. Boar sperm viability in TBM was significantly decreased by 40 and 20% after pretreatment with LY294002 or wortmannin, respectively. Similar results were observed after incubation of boar spermatozoa in TCM. Treatment of boar spermatozoa with the analog of cAMP, 8Br-cAMP significantly prevented the reduction on sperm viability. Our results provide evidence for an important role of the PI3-K pathway in the regulation of boar sperm viability and suggests that other signaling pathways different from PI3-K must be activated downstream of cAMP to contribute to regulation of sperm viability. Finally, in our conditions the PI3-K pathway seems not related with boar sperm capacitation or AR.
Aparicio
I M
IM
Departamento de Fisiología, Universidad de Extremadura, Cáceres, Spain.
Bragado
M J
MJ
Gil
M C
MC
Garcia-Herreros
M
M
Gonzalez-Fernandez
L
L
Tapia
J A
JA
Garcia-Marin
L J
LJ
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000378:metabolism; D000730:Androstadienes / Q000378:metabolism; D000818:Animals; D002470:Cell Survival / Q000502:physiology*; D002867:Chromones / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism; D006801:Humans; D008297:Male; D009025:Morpholines / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D047428:Protein Kinase Inhibitors / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D013075:Sperm Capacitation; D013094:Spermatozoa / Q000502:physiology*; D034421:Sus scrofa; D000077191:Wortmannin
D015124:8-Bromo Cyclic Adenosine Monophosphate; D000730:Androstadienes; D002867:Chromones; D000242:Cyclic AMP; D009025:Morpholines; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D047428:Protein Kinase Inhibitors; D000077191:Wortmannin; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one