Gopal K V, Gross G W
Department of Speech and Hearing Sciences, University of North Texas, Denton 76203, USA.
Acta Otolaryngol. 1996 Sep;116(5):697-704. doi: 10.3109/00016489609137909.
Dissociated embryonic tissue from murine auditory cortex formed spontaneously active monolayer networks in culture that were maintained for up to 113 days in vitro (div). As a first step in determining whether neurons retain histiotypic properties, we subjected a set of 10 cultures to a sequence of 4 synaptically active substances. The test sequence consisted of 50 microM bicuculline, 10 microM strychnine, 5 microM NMDA, and 20 microM GABA. Recordings were made for 5-30 min under each condition followed by complete medium changes. Six to 14 channels with the best signal-to-noise ratios were selected for analysis that consisted of continual chart recordings of integrated burst data and further analysis of short data segments after digitizing and processing. All networks showed spontaneous activity, but had greatly varying native activity ranging from organized, quasi-periodic bursting on all channels to more complex spatio-temporal patterns with less coordination among channels. Bicuculline triggered oscillatory activity, simplified bursting, increased burst amplitude, and enhanced burst regularity among electrodes. Strychnine also changed the burst activity to a simpler pattern and enhanced the burst amplitude, indicating presence of glycine receptors in cortical tissue. Application of NMDA increased burst frequencies, but reduced burst regularity and coordination among channels. 20 microM of GABA inhibited all bursting activity in the networks. These results suggest that monolayer networks cultured on multi-electrode arrays retain some basic histiotypic pharmacological responses and may provide useful platforms for the study of network dynamics in the auditory cortex.
来自小鼠听觉皮层的解离胚胎组织在培养物中形成了自发活动的单层网络,这些网络在体外可维持长达113天。作为确定神经元是否保留组织型特性的第一步,我们对一组10个培养物施加了一系列4种突触活性物质。测试序列包括50微摩尔荷包牡丹碱、10微摩尔士的宁、5微摩尔N-甲基-D-天冬氨酸和20微摩尔γ-氨基丁酸。在每种条件下记录5 - 30分钟,然后完全更换培养基。选择6至14个信噪比最佳的通道进行分析,分析包括对整合爆发数据的连续图表记录以及数字化和处理后对短数据段的进一步分析。所有网络均显示出自发活动,但固有活动差异很大,范围从所有通道上有组织的、准周期性爆发到通道间协调性较低的更复杂时空模式。荷包牡丹碱引发振荡活动、简化爆发、增加爆发幅度并增强电极间爆发规律性。士的宁也将爆发活动改变为更简单的模式并增强了爆发幅度,表明皮质组织中存在甘氨酸受体。应用N-甲基-D-天冬氨酸增加了爆发频率,但降低了爆发规律性和通道间协调性。20微摩尔γ-氨基丁酸抑制了网络中的所有爆发活动。这些结果表明,在多电极阵列上培养的单层网络保留了一些基本的组织型药理学反应,可能为研究听觉皮层的网络动力学提供有用的平台。