Department of Physics, Korea University, Seoul, Korea.
Center for Molecular Spectroscopy and Dynamics, Institute for Basic Science, Seoul, Korea.
Eur J Neurosci. 2019 Sep;50(5):2814-2829. doi: 10.1111/ejn.14417. Epub 2019 Jun 11.
Action potential (AP)-mediated cell-to-cell communication is essential for the frequency-locking and phase-synchronization of the clock cells within the biological master clock, suprachiasmatic nucleus (SCN). Nevertheless, the morphology of its network connectivity is largely unexplored. Here, with an optimized optogenetic light-stimulation and scanning protocol, we report some key characteristics of the inhibitory receptive field (IRF), the area which brings inhibitory synaptic currents to a given target cell, and basic statistics of the inhibitory network connections of rat SCN clock cells. ChR2 transfected, slice cultures of rat SCN were stimulated by a blue power LED light in a repetitive box-scanning modes, while a target cell was whole-cell patched. The registered inhibitory postsynaptic currents, which were brought by light-induced APs of presynaptic neurons, were mostly GABAergic. The sizes and shapes of IRFs of SCN cells were very diverse, and the number of presynaptic cells making up the IRF of a given target cell followed an exponential distribution with an average value of 8.9 approximately, according to our clustering analysis which is based on a hybrid measure D, combining the physical distance r and the difference in the current amplitudes of two different sites. Although this estimate inevitably depends on the construct of the measure D, it is found not so sensitive on the parameter w, which weighs the relative significance of the current amplitude different with respect to the physical distance r: The average number of presynaptic neurons varies < 26% over a significant range of 0 < w < 30. On average, the presynaptic connection number density around a target cell falls off as an exponentially decreasing function of r. But, its space constant (~210.7 μm) is quite large that long-range (>210.7 μm) neural connections are abundant (>66.9%) within the SCN.
动作电位 (AP) 介导的细胞间通讯对于生物主时钟视交叉上核 (SCN) 内时钟细胞的频率锁定和相位同步至关重要。然而,其网络连接的形态在很大程度上仍未得到探索。在这里,我们采用优化的光遗传学光刺激和扫描方案,报告了大鼠 SCN 时钟细胞抑制性感受野 (IRF) 的一些关键特征,IRF 是使给定靶细胞产生抑制性突触电流的区域,以及大鼠 SCN 时钟细胞抑制性网络连接的基本统计数据。用蓝色功率 LED 光以重复框扫描模式刺激转染了 ChR2 的大鼠 SCN 切片培养物,同时对靶细胞进行全细胞贴附。由光诱导的前神经元 AP 引起的登记的抑制性突触后电流主要是 GABA 能的。SCN 细胞的 IRF 大小和形状非常多样化,构成给定靶细胞的 IRF 的前神经元数量遵循指数分布,平均值约为 8.9,根据我们的聚类分析,该分析基于混合度量 D,该度量结合了物理距离 r 和两个不同位点的电流幅度差异。尽管这种估计不可避免地取决于度量 D 的结构,但发现它对参数 w 不那么敏感,w 对电流幅度相对于物理距离 r 的相对重要性进行加权:在前神经元数量在 0 到 30 之间的显著范围内变化<26%。平均而言,靶细胞周围的前突触连接数密度随着 r 的指数衰减函数而下降。但是,它的空间常数(~210.7μm)相当大,以至于 SCN 内存在大量长程(>210.7μm)神经连接(>66.9%)。