Summary
A study of motility was carried out on spermatozoa of the Siberian sturgeon (Acipenser baerii Brandt) reared at the Cemagref experimental research station, St Seurin sur l'Isle, France. About 90% spermatozoa are activated within a few seconds after a double step dilution (final dilution 1/600) and the number of motile cells declines regularly to 10–20% after 2 mn. During the same period of time, the flagellum beat frequency remains stable at about 50 Hz for 30 sec and drops to 30 Hz after 60 sec. A morphological analysis of the movement of the flagellum shows that during the first 20 sec, waves are distributed on the entire length of the flagellum; later, they disappear progressively from the distal part within 1 mn in the case of 90% of the spermatozoa. Finally after 2 mn, most of the flagella are straight and devoid of wave. The ATP content declines very rapidly after activation from 7.58 ± 1.85 to 4.05 ± 1.51 nmol/108 spermatozoa within 5 sec (P=0.004). Spermatozoa left in the genital tract of the males over a period of 24 hrs (in vivo survival) do not show significant change in the percentage of motile cells (82 ± 14 vs 74 ± 15) nor in the ATP content (6.13 ± 2.37 vs 4.59 ± 1.42 nmoles/108 spermatozoa). An aliquot of these spermatozoa stripped from testis and stored during the same period of 24 hrs at 4°C shows a significant decrease in the percentage of motile cells from 82 ± 14 to 59 ± 30 (P<0.01) and in the ATP content from 6.15 ± 2.4 to 1.95 ± 1.37 nmoles/108 spermatozoa (P=0.001).
Live trout spermatozoa initiate flagellar motility for a short period of time (30 s at 18 degrees C), during which their mean beat frequency (BF) decreases steadily from 60 to 20 Hz; motility then stops abruptly. When demembranated, the motility of axonemes lasts much longer, up to 20 min, with high beat frequency, provided that ATP (millimolar concentration) and cAMP (micromolar) are added. In the present paper, the motility of demembranated trout sperm was investigated in the absence of added ATP in various incubation conditions relative to other substrates. Without the addition of exogenous creatine kinase, the addition of phosphocreatine (PCr) and ADP shows the appearance of a progressive activation of all sperm models with BF increasing with time up to high values. Without the addition of cAMP, the BF increases to lower values but flagella propagated poorly coordinated waves for only a few min. Similar progressive activation is also observed when only ADP is added (without any previous in vivo activation) and BF increases up to moderate values. In this latter case, no activation occurs without addition of cAMP. The respective roles of creatine kinase and adenylate kinase in this process were investigated by addition of specific inhibitors such as fluorodinitrobenzene and P1,P5-di(adenosine-5')pentaphosphate in the above described conditions. We conclude from these observations that all the elements necessary for a coupling between ADP/PCr/creatine kinase on one hand and ATP/ADP/dynein on the other appear to be present in trout spermatozoa: thus the existence of a shuttle sustaining this coupling is strongly suggested.
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