ISSN:
0886-1544
Keywords:
Quin-/
;
spermatozoa
;
desmethoxyverapamil
;
fluorescence
;
K+
;
flagellar beating
;
Life and Medical Sciences
;
Cell & Developmental Biology
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Biology
,
Medicine
Notes:
The initiation of motility of trout spermatozoa is inhibited by the presence of millimolar concentrations of external K+, but external Ca2+ might also be implicated in this control as it has been shown to antagonize the K+ inhibition of motility [S.M. Baynes et al.: J. Fish. Biol., 19:259-267, 1981]. The present work aimed to investigate internal Ca2+ levels during the motility phase of trout spermatozoa. Internal Ca2+ concentrations were monitored by the fluorescent quinoline Ca2+-indicator, “Quin-2” [R. Y. Tsien: Nature 290:527-529, 1981]. Trout spermatozoa were loaded with Quin-/ under conditions that gave efficient intracellular hydrolysis of Quin-2 and that did not impair the ability of loaded spermatozoa to initiate movement. The beat frequencies, cell velocities, and flagellar asymmetries of sperm movement were not significantly modified by the presence of the internal dye. Upon initiation of flagellar movement, an increase of the internal Quin-2 fluorescence was observed that reflected a sixfold increase of the free Ca2+ concentration. The free Ca2+ remained elevated after the cessation of movement. The variation of fluorescence was completed within 40 seconds, whereas the initiation of motility was nearly instantaneous, and the total duration of flageliar beating lasted for about 80-100 seconds (measurements at 11°C). The increase in the internal free Ca2+ concentration is completed after the initiation of flagellar beating but its occurrence correlates with that of sperm movement. Fluorescence increase was not observed in the presence of 40 mM K+, a condition in which spermatozoa did not initiate flagellar beating. In the presence of the Ca2+ channel blocker desmethoxyverapamil, neither sperm motility nor fluorescence increases were observed, which suggested that the increase of internal free Ca2+ was produced by a flux of external Ca2+ into the cell rather than by a mobilization of internal Ca2+ stores.
Additional Material:
4 Ill.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/cm.970140312
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