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小龙虾牵张感受器:用电压钳和离子敏感电极进行的研究

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

作者信息

Brown H M, Ottoson D, Rydqvist B

出版信息

J Physiol. 1978 Nov;284:155-79. doi: 10.1113/jphysiol.1978.sp012533.

Abstract
  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.
摘要
  1. 运用膜电位记录、电压钳和离子敏感微电极等电生理技术,对小龙虾(Astacus fluviatilis)的慢适应性牵张感受器的膜特性进行了研究。2. 膜电位阶跃变化至高于0 mV后,被动膜电流(Ip)需超过1分钟才能衰减至稳态水平。3. 牵张诱导电流(SIC,其中SIC = Itotal - Ipassive)直到Ip衰减至稳态时才完全形成。4. 当Ip处于稳态且牵张诱导电流处于零电流电位时,分离出了一种缓慢的牵张诱导内向电流。后者在牵张1秒后达到最大值,牵张后下降得更慢。慢电流的I-V关系具有负斜率,且在静息电位附近符号反转。推测该电流是由于Cl-电导变化所致。5. 在对应于静息电位的钳制电位下,可从缓慢的牵张诱导电流中分离出由快速瞬态相和稳定成分组成的牵张诱导电流。6. SIC-Em关系是非线性的,且在约+15 mV时符号反转。7. 在给定细胞中,电流钳获得的牵张诱导电位变化的反转电位与电压钳获得的牵张诱导电流的零电流电位一致。26个细胞的平均值为+13±6.5 mV;细胞间的变异性似乎与树突长度相关。8. 用Tris(分子量121)或精氨酸(分子量174)替代Na+会降低但不会消除牵张诱导电流。9. 当这些阳离子在外部介质中替代Na+时,根据零电流电位的变化估算了Tris:Na和精氨酸:Na的渗透率比。PTris:PNa略高于精氨酸:PNa的比率(0.25)(为0.31)。10. 外部Ca2+浓度的变化对Na或Tris盐溶液中的零电流电位没有影响。然而,降低Ca2+确实会增加两种盐溶液中的牵张诱导电流。Ca2+浓度降低10倍会使(零电流水平下的)电导增加约两倍。11. 用离子敏感电极获得了细胞内K+和Cl-的活性。六个细胞的平均值为aiK = 133±34 mM,aiCl = 15.2±1.8 mM(标准差)。EK比Em约负20 mV,ECl比Em约正10 mV。12. 在无K+溶液中,aik和静息Em会发生很大变化。60分钟后,ak降低了八倍,Em从-67 mV降至-40 mV。无K+溶液中Ca2+的减少会加快这些变化的速率。无K+溶液中感受器电位幅度也会降低,但在将膜极化至先前的静息水平后可恢复。

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