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Crayfish stretch receptor: an investigation with voltage-clamp and ion-sensitive electrodes.

机译:小龙虾拉伸受体:使用电压钳和离子敏感电极的研究。

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摘要

1. The membrane characteristics of the slowly adapting stretch receptor from the crayfish, Astacus fluviatilis, were examined with electrophysiological techniques consisting of membrane potential recording, voltage clamp and ion-sensitive microelectrodes. 2. The passive membrane current (Ip) following step changes of the membrane potential to levels above 0 mV required more than a minute to decay to a steady-state level. 3. The stretch-induced current (SIC, where SIC = Itotal--Ipassive) was not fully developed until the Ip had decayed to a steady state. 4. With Ip at the steady state and the stretch-induced current at the O-current potential, a slow stretch-induced inward current was isolated. The latter reaches a maximum after 1 sec of stretch and declines even more slowly after stretch. The I-V relation of the slow current had a negative slope and reversed sign near the resting potential. It is suggested that this current is due to a Cl- conductance change. 5. The stretch-induced current, consisting of a rapid transient phase and a steady component can be isolated from the slow stretch-induced current at a holding potential corresponding to the resting potential. 6. The SIC-Em relation is non-linear and reverses sign at about +15 mV. 7. In a given cell, the reversal potential of the stretch-induced potential change obtained with current clamp coincided with the 0-current potential of the stretch-induced current obtained by voltage clamp. The average value from twenty-six cells was +13 +/- 6.5 mV; cell to cell variability seemed to be correlated with dendrite length. 8. Tris (mol. wt. 121) or arginine (mol. wt. 174) susbstituted for Na+ reduces but does not abolish the stretch-induced current. 9. The permeability ratios of Tris:Na and arginine:Na were estimated from changes in the 0-current potential as these cations replaced Na+ in the external medium. The PTris:PNa was somewhat higher (0.31) than the Parginine:PNa ratio (0.25). 10. Changes in the external Ca2+ concentration had no effect on the 0-current potential in Na or Tris saline. However, reducing Ca2+ did augment the stretch-induced current in either saline. A tenfold reduction of Ca2+ increased the conductance (at the 0-current level) about twofold. 11. Intracellular K+ and Cl- activities were obtained with ion sensitive electrodes. The average values from six cells were aiK = 133 +/- 34 mM and aiCl = 15.2 +/- 1.8 mM S.D.). EK was about 20 mV more negative than Em and ECl was about 10 mV more positive than Em. 12. aik and resting Em undergo large changes in K+-free solutions. After 60 min, ak was reduced eightfold and Em was reduced from -67 to -40 mV. Reduced Ca2+ in K+-free augments the rate of these changes. Receptor potential amplitude was also reduced in K+-free solution but could be restored upon polarizing the membrane to the pre-existing resting level.
机译:1.用包括膜电位记录,电压钳和离子敏感微电极在内的电生理技术检查了小龙虾缓慢生长的拉伸受体的膜特性。 2.在膜电位逐步变化到高于0 mV的水平之后,无源膜电流(Ip)需要一分钟以上的时间才能衰减到稳态水平。 3.在Ip衰减到稳态之前,拉伸感应电流(SIC,其中SIC = Itotal-Ipassive)尚未完全形成。 4.在Ip处于稳定状态且拉伸感应电流处于O电流电位的情况下,隔离了缓慢的拉伸感应向内电流。后者在拉伸1秒后达到最大值,在拉伸后下降得更慢。慢电流的I-V关系在静止电位附近具有负斜率和相反的符号。建议该电流是由于Cl电导的变化而引起的。 5.由快速瞬态相位和稳定分量组成的拉伸感应电流可以与慢速拉伸感应电流隔离,保持电流对应于静止电位。 6. SIC-Em关系是非线性的,并且在大约+15 mV时反转符号。 7.在给定的单元中,通过电流钳位获得的拉伸感应电势变化的反向电势与通过电压钳位获得的拉伸感应电流的0电流电势重合。来自二十六个电池的平均值为+13 +/- 6.5 mV;细胞间变异性似乎与枝晶长度有关。 8.替代Na +的Tris(摩尔重量121)或精氨酸(摩尔重量174)降低但不会消除拉伸感应电流。 9. Tris:Na和精氨酸:Na的渗透率比率是根据零电流电位的变化估算的,因为这些阳离子取代了外部介质中的Na +。 PTris:PNa高于(Parginine:PNa)比例(0.25)(0.31)。 10.外部Ca2 +浓度的变化对Na或Tris盐水中的0电流电位没有影响。但是,减少Ca2 +确实会增加两种盐水中的拉伸诱导电流。 Ca2 +降低十倍,电导率(在0电流水平下)增加大约两倍。 11.用离子敏感电极获得细胞内K +和Cl-活性。来自六个电池的平均值为aiK = 133 +/- 34 mM和aiCl = 15.2 +/- 1.8 mM S.D.)。 EK的负电性比Em高20 mV,EC1的正电性比Em高10 mV。 12. aik和静止的Em在无K +的解决方案中发生了很大的变化。 60分钟后,ak减小了八倍,Em从-67减小到-40 mV。不含K +的Ca2 +减少会增加这些变化的速率。在无K +的溶液中,受体电势振幅也降低了,但是在将膜极化到预先存在的静止水平时可以恢复。

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