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钾电导阻断期间新纹状体神经元的被动特性

Passive properties of neostriatal neurons during potassium conductance blockade.

作者信息

Reyes A, Galarraga E, Flores-Hernández J, Tapia D, Bargas J

机构信息

Dept. Biofísica, Instituto de Fisiología Celular, UNAM, Mexico DF, Mexico.

出版信息

Exp Brain Res. 1998 May;120(1):70-84. doi: 10.1007/s002210050379.

Abstract

Voltage recordings from neostriatal projection neurons were obtained using in vitro intracellular techniques before and during K+-conductance blockade. Neurons were stained with the biocytin technique. Somatic surface area (AS) was determined by both whole-cell recordings in isolated somata and by measuring stained somata recorded in slices. Dendritic measurements were done in reconstructed neurons. Average determinations of dendritic (AD) and neuronal (AN) surface areas coincided with previously reported anatomical data. Thus: As approximately 6.5 x 10(-6) cm2; AD approximately 1.9 x 10(-4) cm2; AN approximately AD + AS approximately 2 x 10(-4) cm2; AD/AS approximately 30. Measurements were done before and after superfusion with K+-conductance blockers (K+-blockers). Cells whose neuronal morphology was not obviously distorted by K+-blockade were chosen for the present study. Electrotonic transients were matched to a somatic shunt equivalent cylinder model adjusted with the generalized correction factor (Fdga) that constrains the parameters for neuronal anatomy. Neuronal input resistance (RN; mean +/- SEM) increased when it was corrected for somatic shunt, from 49 +/- 2 Momega (n = 80) to 179 +/- 7 Momega (n = 32). A difference was also obtained between the slowest time constant, tau0 = 16 +/- 0.9 ms (n = 49), and the dendritic membrane time constant, taumD = 33 +/- 1.6 ms (n = 36). When these electrophysiological measurements were used to calculate AN, the value obtained was similar to the anatomical measurements. Combining anatomical and electrophysiological data, somatic and dendritic input resistances were determined: RD = 182 +/- 7 Momega; Rs (with shunt) = 74 +/- 4 Momega (n = 32). The generalized correction factor, Fdga = 0.91 +/- 0.007 (n = 10), implied a short effective electrotonic length for dendrites: LD = 0.46 +/- 0.014 (n = 32). Saturating concentrations of the K+-blockers tetraethylammonium, Cs+, and Ba2+ increased RN and induced charging curves well fitted by single exponential functions in 56% of neostriatal neurons. Ba2+ greatly decreased the somatic shunt (n = 5): (RN = 216 +/- 21 Momega, tau0 = 46 +/- 2 ms, RD = 239 +/- 25 Momega, and RS = 3.2 +/- 0.5 Gomega), rendering values similar to those obtained with whole-cell recordings (e.g., RN approximately 198 Momega, RS approximately 2.62 Gomega) (n = 52). Cs+ (n = 5) had less effect on the somatic shunt (RN = 115 +/- 19 Momega, tau0 = 49 +/- 13 ms, RS = 161 +/- 8 Momega), although dendritic conductance was equally blocked (RD = 261 +/- 16 Momega). The Cs+-sensitive conductance exhibited inward rectifying properties not displayed by the Ba2+-sensitive conductance, suggesting that Cs+ preferentially acted upon inward rectifier conductances. In contrast, Ba2+ significantly acted upon linear conductances making up the somatic shunt. This suggests a differential action of different K+-blockers on the somato-dendritic membrane, implying a differential distribution of membrane conductances. Another action of K+-blockers, in about 40% of the cells, was to induce dye and probably electrical coupling between neighboring neurons.

摘要

在钾离子电导阻断前后,使用体外细胞内技术获取新纹状体投射神经元的电压记录。采用生物胞素技术对神经元进行染色。通过分离胞体的全细胞记录以及测量切片中染色的胞体来确定胞体表面积(AS)。在重建的神经元中进行树突测量。树突表面积(AD)和神经元表面积(AN)的平均测定结果与先前报道的解剖学数据相符。因此:AS约为6.5×10⁻⁶平方厘米;AD约为1.9×10⁻⁴平方厘米;AN约为AD + AS约为2×10⁻⁴平方厘米;AD/AS约为30。在使用钾离子电导阻断剂(钾离子阻断剂)灌流前后进行测量。本研究选择了神经元形态未因钾离子阻断而明显扭曲的细胞。电紧张瞬变与通过广义校正因子(Fdga)调整的胞体分流等效圆柱体模型相匹配,该因子可约束神经元解剖学参数。校正胞体分流后,神经元输入电阻(RN;平均值±标准误)增加,从49±2兆欧(n = 80)增至179±7兆欧(n = 32)。还获得了最慢时间常数tau0 = 16±0.9毫秒(n = 49)与树突膜时间常数taumD = 33±1.6毫秒(n = 36)之间的差异。当使用这些电生理测量值来计算AN时,得到的值与解剖学测量值相似。结合解剖学和电生理数据,确定了胞体和树突输入电阻:RD = 182±7兆欧;Rs(有分流时) = 74±4兆欧(n = 32)。广义校正因子Fdga = 0.91±0.007(n = 10),这意味着树突的有效电紧张长度较短:LD = 0.46±0.014(n = 32)。钾离子阻断剂四乙铵、Cs⁺和Ba²⁺的饱和浓度增加了RN,并在56%的新纹状体神经元中诱导出符合单指数函数的充电曲线。Ba²⁺显著降低了胞体分流(n = 5):(RN = 216±21兆欧,tau0 = 46±2毫秒,RD = 239±25兆欧,RS = 3.2±0.5吉欧),得到的值与全细胞记录获得的值相似(例如,RN约为198兆欧,RS约为2.62吉欧)(n = 52)。Cs⁺(n = 5)对胞体分流的影响较小(RN = 115±19兆欧,tau0 = 49±13毫秒,RS = 161±8兆欧),尽管树突电导也被同等程度地阻断(RD = 261±16兆欧)。Cs⁺敏感电导表现出Ba²⁺敏感电导未显示的内向整流特性,这表明Cs⁺优先作用于内向整流电导。相比之下,Ba²⁺显著作用于构成胞体分流的线性电导。这表明不同的钾离子阻断剂对胞体 - 树突膜有不同的作用,意味着膜电导的分布存在差异。钾离子阻断剂的另一个作用是,在约40%的细胞中,诱导相邻神经元之间的染料偶联以及可能的电偶联。

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