Caputo Carlo, Bolaños Pura, Gonzalez Adom
Centro de Biofisica y Bioquimica, Instituto Venezolano de Investigaciones Cientificas IVIC, Apartado 21827, Caracas, Venezuela.
J Muscle Res Cell Motil. 2004;25(4-5):315-28. doi: 10.1007/s10974-004-4071-z.
MagFluo-4 fluorescence (Ca2+) transients associated with action potentials were measured in intact muscle fibres, manually dissected from toads ( Leptodactylus insularis ) or enzymatically dissociated from mice. In toads, the decay phase of the Ca2+ transients is described by a single exponential with a time constant ( tau ) of about 7 ms. In mice, a double exponential function with tau 's of 1.5 and 15.5 ms, respectively gives a better fit. In both species the amplitude of Ca2+ transients diminished during repetitive stimulation: in amphibian muscle fibres, the decrease was about 20% with 1 Hz stimulation and 55% at 10 Hz. In mammalian fibres, repetitive stimulation causes a less conspicuous decrease of the transient amplitude: 10% at 1 Hz and 15% at 10 Hz. During tetanic stimulation at 100 Hz the transient amplitude decays to 20% in toad fibres and 40% in mouse fibres. This decrease could be associated with the phenomenon of inactivation of Ca2+ release, described by other authors. Recovery from inactivation, studied by a double stimuli protocol also indicates that in toad fibres the ability to release Ca2+ is abolished to a greater extent than in mouse fibres. In fact the ratio between the amplitudes of the second and first transient, when they are separated by a 10 ms interval, is 0.29 for toad and 0.58 for mouse fibres. In toad fibres, recovery from inactivation, to about 80 % of the initial value, occurs with a tau of 32 ms at 22 degrees C; while in mouse fibres recovery from inactivation is almost complete and occurs with a tau of 36 ms under the same conditions. The results indicate that Ca2+ release in enzymatically dissociated mammalian muscle fibres inactivates to a smaller extent than in intact amphibian muscle fibres.
在从蟾蜍(海岛姬蛙)手动解剖或从小鼠酶解分离得到的完整肌纤维中,测量了与动作电位相关的MagFluo-4荧光(Ca2+)瞬变。在蟾蜍中,Ca2+瞬变的衰减相由一个时间常数(τ)约为7 ms的单指数函数描述。在小鼠中,分别具有1.5 ms和15.5 ms的τ的双指数函数拟合效果更好。在这两个物种中,重复刺激期间Ca2+瞬变的幅度均减小:在两栖动物肌纤维中,1 Hz刺激时下降约20%,10 Hz时下降55%。在哺乳动物纤维中,重复刺激引起的瞬变幅度下降不太明显:1 Hz时为10%,10 Hz时为15%。在100 Hz的强直刺激期间,蟾蜍纤维中的瞬变幅度衰减至20%,小鼠纤维中衰减至40%。这种下降可能与其他作者描述的Ca2+释放失活现象有关。通过双刺激方案研究的失活恢复也表明,在蟾蜍纤维中,Ca2+释放能力的丧失程度比小鼠纤维更大。事实上,当第二个和第一个瞬变之间间隔10 ms时,它们的幅度之比在蟾蜍纤维中为0.29,在小鼠纤维中为0.58。在蟾蜍纤维中,在22℃下,失活恢复到初始值的约80%,τ为32 ms;而在小鼠纤维中,失活恢复几乎完全,在相同条件下τ为36 ms。结果表明,酶解分离的哺乳动物肌纤维中的Ca2+释放失活程度比完整两栖动物肌纤维中的小。