Porcine sperm motility is regulated by serine phosphorylation of the glycogen synthase kinase-3alpha.
Sperm functions are critically controlled through the phosphorylation state of specific proteins. Glycogen synthase kinase-3 (GSK3) is a serine/threonine kinase with two different isoforms (alpha and beta), the enzyme activity of which is inhibited by serine phosphorylation. Recent studies suggest that GSK3 is involved in the control of bovine sperm motility. Our aim was to investigate whether GSK3 is present in porcine spermatozoa and its role in the function of these cells. This work shows that both isoforms of GSK3 are present in whole cell lysates of porcine sperm and are phosphorylated on serine in spermatozoa stimulated with the cAMP analog, 8Br-cAMP. A parallel increase in serine phosphorylation of the isoform GSK3alpha, but not in the isoform GSK3beta, is observed after treatments that also induce a significant increase in porcine sperm velocity parameters. Therefore, a significant positive correlation among straight-line velocity, circular velocity, average velocity, rapid-speed spermatozoa, and GSK3alpha serine phosphorylation levels exists. Inhibition of GSK3 activity by alsterpaullone leads to a significant increase in the percentage of rapid- and medium-speed spermatozoa as well as in all sperm velocity parameters and coefficients. Moreover, pretreatment of porcine spermatozoa with alsterpaullone significantly increased the percentage of capacitated porcine spermatozoa and presents no effect in the number of acrosome-reacted porcine spermatozoa. Our work suggests that the isoform GSK3alpha plays a negative role in the regulation of porcine sperm motility and points out the possibility that sperm motile quality might be modulated according the activity state of GSK3alpha.
Aparicio
I M
IM
Research Group of Intracellular Signalling and Technology of Reproduction, Faculty of Veterinary, University of Extremadura, 10071 Caceres, 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 / Q000494:pharmacology; D000818:Animals; D001552:Benzazepines / Q000494:pharmacology; D015153:Blotting, Western / Q000379:methods; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005434:Flow Cytometry; D038362:Glycogen Synthase Kinase 3 / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000071679:Glycogen Synthase Kinase 3 beta; D006358:Hot Temperature; D007211:Indoles / Q000494:pharmacology; D008297:Male; D008858:Microscopy, Phase-Contrast; D010766:Phosphorylation; D012694:Serine / Q000378:metabolism*; D013081:Sperm Motility / Q000187:drug effects / Q000502:physiology*; D013268:Stimulation, Chemical; D013552:Swine / Q000502:physiology*
D001552:Benzazepines; D004791:Enzyme Inhibitors; D007211:Indoles; C120793:alsterpaullone; D015124:8-Bromo Cyclic Adenosine Monophosphate; D012694:Serine; D000242:Cyclic AMP; D000071679:Glycogen Synthase Kinase 3 beta; D017868:Cyclic AMP-Dependent Protein Kinases; D038362:Glycogen Synthase Kinase 3; C448021:glycogen synthase kinase 3 alpha