Analysis of synaptic inputs and targets of physiologically characterized neurons in rat frontal cortex: combined in vivo intracellular recording and immunolabeling.
Analysis of synaptic inputs and targets of physiologically characterized neurons in rat frontal cortex: combined in vivo intracellular recording and immunolabeling.
复制标题
大鼠额叶皮层生理特征神经元的突触输入和目标分析：体内细胞内记录和免疫标记相结合。
DOI：
10.1002/syn.890170206
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发表时间：
1994
期刊：
影响因子：
--
通讯作者：
Pickel,VM
中科院分区：
文献类型：
--
作者：
Cowan,RL;Sesack,SR;VanBockstaele,EJ;Branchereau,P;Chain,J;Pickel,VM
超微结构的免疫细胞化学识别的传入终端和目标的个人生理特征的神经元的递质是必不可少的了解哺乳动物新皮层内的复杂电路。对于这种类型的分析，我们研究了结合体内细胞内记录和生物胞素注射与银增强1 nm免疫金标记的GABA和儿茶酚胺合成酶，酪氨酸羟化酶（TH）的效用。这些递质存在于局部神经元和传入神经元中，已知其显著调节新皮层中锥体神经元的活动。麻醉大鼠额叶皮层单个神经元具有生理学特征，并充满生物胞素。然后通过醛类的血管灌注保存大脑。通过记录位点的单个振动切片机切片进行反应（1）用于生物胞素的免疫过氧化物酶检测和（2）用于GABA或TH的免疫金标记。处理双重标记的切片用于光学显微镜检查或包埋在塑料中用于电子显微镜检查。在锥体神经元中检测到生物胞素的密集过氧化物酶产物。这些位于浅层和深层皮质层。并且容易与免疫金银标记区分。GABA标记的终末与较大的生物胞素填充的树突形成对称的突触，而TH标记的终末接触远端树突和棘。过氧化物酶标记的biocytin也被认为是在一些轴突终末形成突触与未标记和GABA免疫反应阳性树突。这些结果表明，大鼠额叶皮层单个锥体神经元接受GABA和儿茶酚胺终末的双重输入。此外，这项研究表明，银增强1 nm coilloidal金塑料包埋前的电子显微镜检测神经递质的传入和目标的神经元的生理特征在体内的有用性。© 1994年威利利斯。Inc.
Ultrastructural immunocytochemical identification of transmitters in afferent terminals and targets of individual physiologically characterized neurons is essential for understanding the complex circuitry within the mammalian neocortex. For this type of analysis, we examined the utility of combining in vivo intracellular recording and biocytin injections with silver intensified 1 nm immunogold labeling of GABA and the catecholamine synthesizing enzyme, tyrosine hydroxylase (TH). These transmitters are found in local neurons and afferents known to prominently modulate the activity of pyramidal neurons in the neocortex. Individual neurons were physiologically characterlzea and filled with biocytin in the frontal cortex of anesthetized rats. The brains were then preserved by vascular perfusion with aldehydes. Single vibratome sections through the recording site were reacted (1) for immunoperoxidase detection of biocytin and (2) for immunogold labeling of GABA or TH. Dually labeled sections were processed for light microscopy or embedded in plastic for electron microscopy. The dense peroxidase product for biocytin was detected in pyramidal neurons. These were located in superficial as well as deep cortical laminae. and were readily distinguished from immunogold silver labeling. GABA labeled terminals formed symmetric synapses with larger biocytin filled dendrites, whereas the TH labeled terminals contacted distal dendrites and spines. Peroxidase labeling for biocytin also was seen in a few axon terminals forming synapses with unlabeled and with GABA immunoreactive dendrites. These results suggest that single pyramidal neurons of the rat frontal cortex receive dual input from both GABA and catecholamine terminals. Additionally, this study demonstrates the usefulness of silver enhancement of 1 nm coilloidal gold prior to plastic embedding for electron microscopic detection of neurotransmitters within afferents and targets of neurons physiologically characterized in vivo. © 1994 Wiley‐Liss. Inc.