Changes in sperm glycogen synthase kinase-3 serine phosphorylation and activity accompany motility initiation and stimulation.
Sperm motility is regulated by protein phosphorylation. We have shown that the signaling kinase, glycogen synthase kinase-3 alpha (GSK-3 alpha), is present in spermatozoa. In somatic cells, GSK-3 is regulated by serine and tyrosine phosphorylation. In this report, we document that both GSK-3 alpha and GSK-beta isoforms are present in spermatozoa, with GSK-3 alpha being the predominant isoform. The relationship between GSK-3 serine phosphorylation and motility was investigated. Serine phosphorylation of GSK-3 increases significantly in spermatozoa during their passage through the epididymis. Initiation and stimulation of motility in vitro by isobutyl-methyl-xanthine, 2-chloro-2'-deoxy-adenosine, and calyculin A lead to a dramatic increase in GSK-3 serine phosphorylation. The concentration-dependent induction of motility by calyculin A is closely associated with GSK-3 serine phosphorylation. Immunoprecipitation of GSK-3 alpha and GSK-3 beta shows that both of the GSK-3 isoforms are more active in caput than in caudal spermatozoa. Calyculin A treatment decreased the activity of both isoforms. Column chromatography was used to purify inactive GSK-3 alpha from the caudal sperm extracts. This GSK-3 alpha species was phosphorylated at amino acid residues serine 21 and tyrosine 214. Inactive GSK-3 alpha is present in caudal but not in caput epididymal spermatozoa. The enzymes protein kinase B (PKB; also known as cAkt) and phosphoinositide 3-kinase (PI3-kinase), the upstream signaling proteins involved in GSK-3 phosphorylation, are both present in spermatozoa. Fluorescence immunocytochemistry showed that GSK-3 is present in the head and tail regions of sperm. Our work suggests a novel role for the signaling system involving GSK-3 in the regulation of sperm motility.
Somanath
Payaningal R
PR
Department of Biological Sciences, Kent State University, Kent, Ohio 44242, USA.
Jack
Shannan L
SL
Vijayaraghavan
Srinivasan
S
D000818:Animals; D015153:Blotting, Western; D002417:Cattle; D038362:Glycogen Synthase Kinase 3 / Q000302:isolation & purification / Q000378:metabolism*; D008297:Male; D010766:Phosphorylation; D010768:Phosphoserine / Q000378:metabolism*; D013081:Sperm Motility / Q000502:physiology*; D013094:Spermatozoa / Q000201:enzymology*
D010768:Phosphoserine; D038362:Glycogen Synthase Kinase 3