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对蜥蜴和青蛙视网膜视杆细胞中光敏暗噪声的膜片钳记录。

Patch-clamp recordings of the light-sensitive dark noise in retinal rods from the lizard and frog.

作者信息

Bodoia R D, Detwiler P B

出版信息

J Physiol. 1985 Oct;367:183-216. doi: 10.1113/jphysiol.1985.sp015820.

Abstract

In cell-attached recordings from rods in the intact lizard retina, light decreased a standing inward membrane current with a reversal potential approximately 60 mV more positive than the resting potential. The peak amplitude of saturating responses depended upon the area of recorded membrane and varied from cell to cell over approximately 100-fold range. Small patches of membrane gave variable responses to identical moderately intense flashes. Whole-cell voltage-clamp recordings were obtained on isolated frog rods with intact ellipsoids. Peak whole-cell photocurrent was related to flash intensity by a Michaelis equation with saturating response amplitudes ranging up to 30 pA in 0.1 mM-Ca2+ Ringer solution. In darkness the steady-state current-voltage relation, determined with whole-cell voltage clamp, showed outward rectification. Photocurrent had nearly constant amplitude between -80 and -10 mV, a mean reversal potential of +8 mV and recovered from flashes more slowly at positive holding potentials. Although it was not possible to resolve light-sensitive single-channel current events, power spectral analysis revealed both low- and high-frequency components of the light-sensitive noise in both cell-attached and whole-cell recordings. The low-frequency component was described by the product of two Lorentzians using time constants derived from the kinetics of the dim flash response. The high-frequency component of the light-sensitive noise was described by a single Lorentzian with a half-power frequency of 62 Hz in lizard and 212 Hz in frog. The half-power frequency was not appreciably affected by steady illumination. The Lorentzian nature of the noise suggests that the light-sensitive channel is a pore rather than a shuttle-type carrier. In cell-attached recordings the high-frequency component declined monotonically with increasing light intensity, suggesting that less than one-half of the channels are open in darkness. Furthermore, the ratio of the variance of the high-frequency noise to the mean photocurrent was independent of light intensity. Changing external Ca2+ from 0.1 to 0.5 mM reduced the ratio from 19.7 to 9.0 fA without a significant effect on the cut-off frequency of the noise. The results support the conclusion that the light-sensitive pore is opened by an internal transmitter that acts as an agonist and that both open and closed states of the pore may be blocked by external Ca2+. The conductance of the light-sensitive pore in the absence of external Ca2+ is estimated to be 1.25-2 pS.(ABSTRACT TRUNCATED AT 400 WORDS)

摘要

在完整蜥蜴视网膜视杆细胞的细胞贴附式记录中,光照使内向性膜电流减小,其反转电位比静息电位正约60 mV。饱和反应的峰值幅度取决于记录膜的面积,细胞间变化范围约为100倍。小面积膜对相同强度的中等闪光产生可变反应。在具有完整椭球体的分离青蛙视杆细胞上进行全细胞膜片钳记录。在0.1 mM - Ca2+林格氏液中,全细胞光电流峰值与闪光强度符合米氏方程,饱和反应幅度高达30 pA。在黑暗中,用全细胞膜片钳测定的稳态电流 - 电压关系显示外向整流。光电流在 - 80至 - 10 mV之间幅度几乎恒定,平均反转电位为 + 8 mV,在正的钳制电位下从闪光恢复得更慢。虽然无法分辨光敏感单通道电流事件,但功率谱分析在细胞贴附式和全细胞记录中均揭示了光敏感噪声的低频和高频成分。低频成分由两个洛伦兹函数的乘积描述,使用从暗闪光反应动力学得出的时间常数。光敏感噪声的高频成分由单个洛伦兹函数描述,蜥蜴中半功率频率为62 Hz,青蛙中为212 Hz。半功率频率不受持续光照明显影响。噪声的洛伦兹性质表明光敏感通道是一个孔而不是穿梭型载体。在细胞贴附式记录中,高频成分随光照强度增加单调下降,表明在黑暗中不到一半的通道开放。此外,高频噪声方差与平均光电流的比值与光照强度无关。将外部Ca2+从0.1 mM变为0.5 mM可使该比值从19.7降至9.0 fA,而对噪声截止频率无显著影响。结果支持以下结论:光敏感孔由作为激动剂的内部递质打开,孔的开放和关闭状态均可被外部Ca2+阻断。在无外部Ca2+时,光敏感孔的电导估计为1.25 - 2 pS。(摘要截断于400字)

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/13eb/1193059/bed206ebd813/jphysiol00565-0227-a.jpg

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