Development of three-dimensional optical recording methods for neural activity in the central nervous system.

为了评估早期胚胎鸡脑干舌咽反应的空间模式，我们使用了一个12×12元光电二极管阵列的多点光学记录系统和电压敏感的花青红-罗丹宁染料来监测神经跨膜电压活动。取7日龄和8日龄鸡胚脑切片，厚片1400~1600微米，分别连接舌咽和迷走神经，染色后取材。在复合显微镜的物镜二维像平面上，通过微抽吸电极对舌咽神经施加短时(去极化)正方电流脉冲，同时记录下神经电压相关的光信号。除了多点光学切片方法外，还可以通过改变显微镜的焦平面来获得三维信息。因此，我们已经能够半定量地评估对应于舌咽神经核(舌咽神经核)和溶质束核的两个舌咽反应区的三维轮廓。此外，还在孤束核对应的反应区内测定了谷氨酸能兴奋性突触后电位。此外，我们还比较了舌咽和迷走神经反应区，发现相关核团的核心在三维上是分开的。

In an effort to asses the spatial patterning of glossopharyngeal responses in early embryonic chick brainstem, we used a multiple-site optical recording system with a 12*12 element photodiode array and voltage sensitive merocyanine-rhodanine dyes to monitor neural transmembrane voltage activities. Seven and 8 d old embryonic chick brainstems were sliced into 1400-1600 mum thick sectons with the glossopharyngeal and vagal nerves attached, and then attained withthe dye. Neural voltage-related optical signals were evoked by a positive brief (depolarizing) square current pulse applied to the glossopharyngeal nerve with a microsuction electrode, and then recorded simultaneously from many loci in the objective two-dimensional image plane of a compound microscope. In addition to the multiple-site optical sectioning method by changing the focal plane of the microscope to obtain three-dimensional information. Thus, we have been able to assess semiquantitatively the three-dimensional profiles of two glossopharyngeal response areas corresponding to the nucleus of the glossopharyngeal nerve (nucleus nervi glossopharyngei) and the necleus of the tractus solutarius. Furthermore, glutaminergic excitatory postsynaptic potentials were determined within the response area corresponding to the nucleus of the tractus solitarius. In additon, we also compared the glossopharyngesl and vagal response areas and found that the cores of the related nuclei are separated in three dimensions.