Terakawa S
J Physiol. 1985 Dec;369:229-48. doi: 10.1113/jphysiol.1985.sp015898.
Intracellular pressure responses were recorded from squid giant axons after the axoplasm was removed by the intracellular perfusion technique. A glass tube was inserted into the axon and the movement of the air-water interface formed on the end of the tube was observed with a Y-shaped fibrescope. The intracellular pressure increased and decreased rapidly when an action potential was induced by electrical stimulation. The amplitude of the response was about 10 mPa (or 1 X 10(-3) mmH2O), which was very large in comparison with that observed in unperfused axons. It was sensitive to extracellular Ca2+. The pressure response appeared in an all-or-none manner and could be suppressed by tetrodotoxin. This excluded physicochemical processes on the stimulating electrode or current-supplying electrode as sources of the response. Various other sources of artifacts were also excluded. An extensive removal of the axoplasm by intracellular perfusion with a protease-containing solution and a KCl solution did not eliminate the pressure response. The intracellular pressure was membrane potential dependent, increasing upon depolarization and decreasing upon hyperpolarization of the membrane. Under voltage clamp, the relationship between the membrane potential and the pressure response was parabolic with a maximum at +109 mV (in reference to the resting level). The response did not depend on the membrane current. A much slower response due to electro-osmotic water flow was also detected. The pressure response induced by hyperpolarization of the membrane was suppressed by extracellular application of a lidocaine-containing solution, but not by a tetrodotoxin-containing solution. These results suggest that the pressure responses arise either from a change in electrostriction across the axolemma or from a change in charge-dependent tension along the axolemma.
采用细胞内灌注技术去除轴浆后,记录枪乌贼巨大轴突的细胞内压力反应。将一根玻璃管插入轴突,用Y形纤维镜观察在管端形成的气-水界面的移动。当通过电刺激诱发动作电位时,细胞内压力迅速升高和降低。反应幅度约为10 mPa(或1×10⁻³ mmHg₂O),与未灌注轴突中观察到的相比非常大。它对细胞外Ca²⁺敏感。压力反应以全或无的方式出现,并且可被河豚毒素抑制。这排除了刺激电极或供电电极上的物理化学过程作为反应源。还排除了各种其他伪迹来源。用含蛋白酶的溶液和KCl溶液进行细胞内灌注广泛去除轴浆并不能消除压力反应。细胞内压力取决于膜电位,在膜去极化时升高,在膜超极化时降低。在电压钳制下,膜电位与压力反应之间的关系呈抛物线形,在+109 mV(相对于静息水平)时达到最大值。反应不依赖于膜电流。还检测到由于电渗水流引起的慢得多的反应。膜超极化诱导的压力反应可被细胞外应用含利多卡因的溶液抑制,但不能被含河豚毒素的溶液抑制。这些结果表明,压力反应要么源于轴膜上电致伸缩的变化,要么源于沿轴膜上电荷依赖性张力的变化。