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Analysis of synaptic formation on the cerebral pyramidal cell associated with the projection of thalamocortical axons

大脑锥体细胞突触形成与丘脑皮质轴突投射相关的分析

基本信息

批准号：
09680736
负责人：
ISHII Katsuyoushi
金额：
$ 1.86万
依托单位：
The institute of Physical and Chemical Research (RIKEN) (1998)Yamagata University (1997)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

I examined the outgrowth of thalamocortical axons in the developing rat and their distribution in the cerebral cortex with DiI and Biocytin labeling methods. These findings showed the morphology of thalamocortical axons as they arrive at the intermediate zone at embryonic day(ED) 16, as they enter into the deep layer of the cerebral cortex at ED18, and begin to make terminal branching in developing layer IV at postnatal day(PD)2 and form more clearly periodic and dense clusters by PD7.To obtain well-labeled GABAergic neurons in layer IV of the cerebral cortex for morphological analysis, I injected dye into GABA-immunopositive neurons in fixed slices. I injected Biocytin from a posterior direction into the right thalpmus of rats and labeled thalamocortical axons with Biocytin anterogradely. Sections were reacted for GABA immunohistochemistry and revealed GABA positive neurons with FITC. So I injected Lucifer yellow intracellularly into FITC labeled cells in layer IV of the cerebral cortex. Injected cells were imaged with a laser conforcal microscope. Most injected neurons were multipolar at all ages. At PD7 or younger injected cells with vertically elongated or polygonal soma and a major dendrite vertically elongated were more common. At PD14 and 21, many injected- cells had spherical or oval soma and showed more dendrites radiated from the soma in all directions. These findings show the morphology of GABAergic neurons change into mature type between PD7 and PD 14.To study the synaptic formation on the cerebral pyramidal cell associated with the projection of thalamocortical axons, I labeled thalamocortical axons with Biocytin anterogradely and cerebral pyramidal cells with Fluoro-Gold retrogradely. So I injected Lucifer yellow intracellularly into Fluoro-Gold labeled cells in layers V and II/III of the cerebral cortex. Using specimens for an electron microscope, the study of synaptic formation on the cerebral pyramidal cell is in progress.

用DiI和Biocytin标记法观察了发育期大鼠丘脑皮质轴突的生长及其在大脑皮质的分布。这些发现显示了丘脑皮质轴突在胚胎（艾德）16天到达中间区时的形态，当它们在胚胎18天进入大脑皮层深层时，出生后第2天（PD 2），发育层Ⅳ开始开始产生终末分支，到PD 7时形成周期性更明显、密度更大的簇。标记的大脑皮层第IV层GABA能神经元进行形态学分析，将染料注射到固定切片的GABA免疫阳性神经元中。我将Biocytin从后方注射到大鼠的右丘脑，并用Biocytin顺行标记丘脑皮质轴突。切片进行GABA免疫组织化学反应，用FITC显示GABA阳性神经元。所以我将荧光黄注射到大脑皮层第四层的异硫氰酸荧光素标记细胞中。注射的细胞用激光共聚焦显微镜成像。在所有年龄段，大多数注射神经元都是多极的。在PD 7或更年轻时，注射细胞具有垂直伸长或多边形索马和垂直伸长的主要树突更常见。在PD 14和21时，许多注射的细胞具有球形或椭圆形的索马体，并显示出更多的树突从索马体向各个方向放射。结果表明，在PD 7 ~ PD 14之间，GABA能神经元的形态发生了变化，逐渐向成熟型转变。为了研究与丘脑皮质轴突投射相关的大脑锥体细胞上的突触形成，我用Biocytin顺标丘脑皮质轴突，Fluoro-Gold逆标大脑锥体细胞。所以我把荧光黄注射到大脑皮层第五层和第二/第三层的荧光金标记细胞中。利用电镜标本对大脑锥体细胞突触形成的研究正在进行中。