Günay Cengiz, Sieling Fred H, Dharmar Logesh, Lin Wei-Hsiang, Wolfram Verena, Marley Richard, Baines Richard A, Prinz Astrid A
Department of Biology, Emory University, Atlanta, Georgia, United States of America.
Department of Biology, Emory University, Atlanta, Georgia, United States of America; Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom.
PLoS Comput Biol. 2015 May 15;11(5):e1004189. doi: 10.1371/journal.pcbi.1004189. eCollection 2015 May.
Studying ion channel currents generated distally from the recording site is difficult because of artifacts caused by poor space clamp and membrane filtering. A computational model can quantify artifact parameters for correction by simulating the currents only if their exact anatomical location is known. We propose that the same artifacts that confound current recordings can help pinpoint the source of those currents by providing a signature of the neuron's morphology. This method can improve the recording quality of currents initiated at the spike initiation zone (SIZ) that are often distal to the soma in invertebrate neurons. Drosophila being a valuable tool for characterizing ion currents, we estimated the SIZ location and quantified artifacts in an identified motoneuron, aCC/MN1-Ib, by constructing a novel multicompartmental model. Initial simulation of the measured biophysical channel properties in an isopotential Hodgkin-Huxley type neuron model partially replicated firing characteristics. Adding a second distal compartment, which contained spike-generating Na+ and K+ currents, was sufficient to simulate aCC's in vivo activity signature. Matching this signature using a reconstructed morphology predicted that the SIZ is on aCC's primary axon, 70 μm after the most distal dendritic branching point. From SIZ to soma, we observed and quantified selective morphological filtering of fast activating currents. Non-inactivating K+ currents are filtered ∼3 times less and despite their large magnitude at the soma they could be as distal as Na+ currents. The peak of transient component (NaT) of the voltage-activated Na+ current is also filtered more than the magnitude of slower persistent component (NaP), which can contribute to seizures. The corrected NaP/NaT ratio explains the previously observed discrepancy when the same channel is expressed in different cells. In summary, we used an in vivo signature to estimate ion channel location and recording artifacts, which can be applied to other neurons.
由于空间钳制不佳和膜滤波导致的伪迹,研究记录位点远端产生的离子通道电流很困难。只有在电流的确切解剖位置已知的情况下,计算模型才能通过模拟电流来量化伪迹参数以进行校正。我们提出,那些干扰电流记录的相同伪迹可以通过提供神经元形态特征来帮助确定这些电流的来源。这种方法可以提高在无脊椎动物神经元中通常位于胞体远端的峰起始区(SIZ)起始的电流的记录质量。果蝇是表征离子电流的有价值工具,我们通过构建一个新颖的多房室模型,估计了一个已鉴定的运动神经元aCC/MN1-Ib中的SIZ位置并量化了伪迹。在等电位霍奇金-赫胥黎型神经元模型中对测量的生物物理通道特性进行的初步模拟部分复制了放电特征。添加一个包含产生峰电位的Na+和K+电流的第二个远端房室,足以模拟aCC的体内活动特征。使用重建形态匹配该特征预测SIZ位于aCC的初级轴突上,在最远端树突分支点之后70μm处。从SIZ到胞体,我们观察并量化了快速激活电流的选择性形态滤波。非失活K+电流的滤波程度约低3倍,尽管它们在胞体处幅度很大,但它们可以与Na+电流一样远。电压激活Na+电流的瞬态成分(NaT)的峰值也比持续时间较慢的成分(NaP)的幅度滤波更多,后者可能导致癫痫发作。校正后的NaP/NaT比值解释了先前在不同细胞中表达相同通道时观察到的差异。总之,我们使用体内特征来估计离子通道位置和记录伪迹,这可应用于其他神经元。