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

开发中枢神经系统神经活动的三维光学记录方法。

基本信息

批准号：
06558110
负责人：
KAMINO Kohtaro
金额：
$ 11.65万
依托单位：
Tokyo Medical and Dental University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1996
项目状态：
已结题

项目摘要

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.

为了评估早期胚胎鸡脑干舌咽部反应的空间模式，我们使用了一个带有12*12元素光电二极管阵列和电压敏感的merocyanine-rhodanine染料的多位点光学记录系统来监测神经跨膜电压活动。取7、8 d龄胚鸡脑干切片，切成1400 ~ 1600 mm厚，附着舌咽神经和迷走神经，染色。用微吸电极对舌咽神经施加短时间正极（去极化）方电流脉冲，诱发神经电压相关的光信号，并在复合显微镜的物镜二维像面上同时记录多个位点的光信号。另外多位点光学切片法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）和舌束核的两个舌咽部反应区域的三维轮廓。此外，在孤束核对应的反应区域内测定谷氨酰胺能兴奋性突触后电位。此外，我们还比较了舌咽部和迷走神经反应区，发现相关核的核心在三维空间上是分离的。