Acclimation of sperm motility apparatus in seawater-acclimated euryhaline tilapia Oreochromis mossambicus.
Euryhaline tilapia Oreochromis mossambicus can reproduce in freshwater and in seawater. Regulation of sperm motility appears to be modulated during acclimation of the fish from freshwater to seawater, being independent of extracellular Ca2+ in freshwater and dependent on extracellular Ca2+ in seawater. In the presence of extracellular Ca2+, sperm of seawater-acclimated tilapia (SWT) showed motility even in a hypertonic environment, whereas sperm of freshwater-acclimated tilapia (FWT) were not motile. The Ca2+ indicator, fluo-3, revealed that intracellular Ca2+ concentration, [Ca2+]i, of SWT sperm increased only in the presence of extracellular Ca(2+) in hypotonic or hypertonic conditions. Since the increased [Ca2+]i in FWT sperm occurred under hypotonic conditions via intracellular Ca2+ stores, it is likely that tilapia modulate their source of increasing [Ca2+]i from intracellular stores (in FWT sperm) to extracellular stores (in SWT sperm). Experiments using demembranated sperm revealed that Ca2+ is necessary for activation of motility, suggesting that Ca2+ plays a key role in motility regulation in SWT sperm. We detected three phosphoproteins associated with the activation of sperm motility. Serine and threonine residues of two proteins of 15 kDa and 18 kDa became dephosphorylated in hypotonic conditions but remained phosphorylated in hypertonic conditions, suggesting that these protein phosphorylations were not only related to motility activation under hypertonic conditions but also resistant to osmotic pressure. The threonine residue(s) of a 41 kDa protein was also phosphorylated in dry sperm, even in FWT sperm in motility-feasible hypotonic conditions. It is likely that acclimation of the motility apparatus is associated with modulation of the flow of Ca2+ to increase [Ca2+]i and protein phosphorylation.
Morita
Masaya
M
Department of Life Sciences, Graduate School of Arts and Sciences, University of Tokyo, Meguro-ku, Tokyo 153-8902, Japan.
Takemura
Akihiro
A
Okuno
Makoto
M
D000064:Acclimatization / Q000502:physiology*; D000704:Analysis of Variance; D000814:Aniline Compounds; D000818:Animals; D001692:Biological Transport / Q000502:physiology; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism* / Q000502:physiology; D004591:Electrophoresis, Polyacrylamide Gel; D007564:Japan; D008297:Male; D018613:Microscopy, Confocal; D010750:Phosphoproteins / Q000378:metabolism / Q000502:physiology; D010766:Phosphorylation; D012623:Seawater; D012694:Serine / Q000378:metabolism; D013081:Sperm Motility / Q000502:physiology*; D013912:Threonine / Q000378:metabolism; D017210:Tilapia / Q000502:physiology*; D014966:Xanthenes
D000814:Aniline Compounds; D010750:Phosphoproteins; D014966:Xanthenes; C059715:Fluo-3; D013912:Threonine; D012694:Serine; D002118:Calcium