Saudrais C, Fierville F, Loir M, Le Rumeur E, Cibert C, Cosson J
Laboratoire de Physiologie des Poissons, INRA, Campus de Beaulieu, Rennes, France.
Cell Motil Cytoskeleton. 1998;41(2):91-106. doi: 10.1002/(SICI)1097-0169(1998)41:2<91::AID-CM1>3.0.CO;2-I.
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.
活的鳟鱼精子在短时间内（18摄氏度下30秒）启动鞭毛运动，在此期间其平均拍频（BF）从60赫兹稳步下降至20赫兹；然后运动突然停止。去膜后，只要添加ATP（毫摩尔浓度）和cAMP（微摩尔浓度），轴丝的运动持续时间会长得多，可达20分钟，且拍频较高。在本文中，研究了在各种相对于其他底物的孵育条件下，去膜鳟鱼精子在不添加ATP时的运动情况。在不添加外源性肌酸激酶的情况下，添加磷酸肌酸（PCr）和ADP会使所有精子模型逐渐激活，BF随时间增加直至达到较高值。在不添加cAMP的情况下，BF增加到较低值，但鞭毛仅在几分钟内传播协调性较差的波动。当仅添加ADP（无任何先前的体内激活）时也观察到类似的逐渐激活，且BF增加至中等值。在后一种情况下，不添加cAMP则不会发生激活。通过在上述条件下添加氟二硝基苯和P1，P5 - 二（腺苷 - 5'）五磷酸等特异性抑制剂，研究了肌酸激酶和腺苷酸激酶在此过程中的各自作用。我们从这些观察结果得出结论，一方面ADP/PCr/肌酸激酶与另一方面ATP/ADP/动力蛋白之间耦合所需的所有元素似乎都存在于鳟鱼精子中：因此强烈暗示存在维持这种耦合的穿梭机制。
