Identification of different putative neuronal subtypes in cultures of the superior region of the hippocampus using electrophysiological parameters.
Identification of different putative neuronal subtypes in cultures of the superior region of the hippocampus using electrophysiological parameters.
复制标题
使用电生理参数识别海马上部区域培养物中不同的假定神经元亚型。
DOI：
10.1016/s0306-4522(99)00153-0
复制
发表时间：
1999
期刊：
影响因子：
3.3
通讯作者：
Mynlieff,M
中科院分区：
文献类型：
--
作者：
Mynlieff,M
培养的神经元在全细胞膜片钳研究中提供了许多优于切片制备的优势，例如更好地控制环境和空间钳控制。然而，神经元的异质培养在区分进行记录的细胞类型方面存在问题。本研究使用相关性与海马切片制备中获得的数据，以确定“识别”不同的神经元亚型的培养物中获得的出生后2至13天的大鼠海马的上级地区的可行性。在培养24- 96小时后以电流钳模式记录全细胞膜片钳，以确定动作电位时程是否是区分细胞类型的有用标准。用0.1-0.2ms、2- 4 nA去极化脉冲诱发单个动作电位。平均膜电位和输入电阻分别为−46.8±1.2mV（n=58）和576± 56 M Ω（n=57）。在半最大振幅下测量的动作电位时程的频率分布显示四个不同的神经元组（组1，1.36±0.03ms，n=17;组2，2.19±0.05ms，n=20;组3，3.17±0.10ms，n=16;组4，4.36±0.13，n=5）。基于与先前使用切片中识别的细胞的细胞内记录的研究的相关性，数据表明，组1代表篮状细胞，组2代表垂直细胞，组3代表星状细胞和锥体细胞的组合，组4代表另一类未识别的细胞。快速后超极化的进一步分析允许区分锥体细胞和星状细胞组3。与切片制备中的中间神经元相比，这些细胞提供良好的电压控制和环境控制。未来的研究将以电流钳模式记录这些细胞，以快速表征动作电位，然后切换到电压钳记录，以表征不同类型的中间神经元中存在的电流。
Cultured neurons offer many advantages over a slice preparation for whole-cell patch-clamp studies, such as better control over the environment and space clamp control. However, heterogeneous cultures of neurons present problems in distinguishing the cell type from which recordings are made. The present study uses correlations with data obtained in the hippocampal slice preparation to determine the feasibility of “identifying” different neuronal subtypes in cultures obtained from the superior region of postnatal two- to 13-day-old rat hippocampus. Whole-cell patch-clamp recording in the current-clamp mode after 24–96h in culture was used to determine if the action potential duration would be a useful criterion in distinguishing cell types. Single action potentials were elicited by a 0.1–0.2ms, 2–4nA depolarizing pulse. The average membrane potential and input resistance were −46.8±1.2mV (n=58) and 576±56MΩ (n=57), respectively. A frequency distribution of the action potential duration measured at half-maximal amplitude showed four distinct groups of neurons (group 1, 1.36±0.03ms, n=17; group 2, 2.19±0.05ms, n=20; group 3, 3.17±0.10ms, n=16; group 4, 4.36±0.13, n=5). Based on correlations with previous studies using intracellular recording in identified cells in slices, the data suggest that group 1 represents basket cells, group 2 represents vertical cells, group 3 represents a combination of stellate cells and pyramidal cells, and group 4 represents another unidentified class of cells. Further analysis of the fast afterhyperpolarization allows distinction between pyramidal cells and stellate cells in group 3. In contrast to the interneurons in a slice preparation, these cells offer good voltage control and environmental control. Future studies will record from these cells in current-clamp mode to quickly characterize the action potential before switching to voltage-clamp recording to characterize the currents present in the different types of interneurons.