Nitric oxide synthase in rat visual cortex: an immunohistochemical study

Tagrid Yousef , Ute Neubacher , Ulf T Eysel , Maxim Volgushev
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引用次数: 22

Abstract

The aim of the present study was to identify the distribution of two isoforms of the nitric oxide synthase (NOS), the neuronal (nNOS) and the endothelial (eNOS) form, in rat visual cortex. Immunohistochemical localisation of each NOS isoform was studied with three tissue-processing protocols. In the first one, immunohistochemical reactions were made on 30-μm-thick sections with membrane detergents, Triton or Saponin, used to increase the permeability of the tissue for the antibodies. In the second protocol, we excluded these detergents from all solutions to avoid a destruction of the cellular membrane. In the third protocol, we used thin paraffin sections (5 μm thick) to assure delivery of the antibodies to intracellular structures. Our data demonstrate, that both neuronal and endothelial isoforms of the NOS are present in the visual cortex. Among the neurones labelled by the antibodies against eNOS or nNOS, some excitatory cells were definitely present. nNOS immunopositive were neurones and a dense network of fibres, presumably axons. Some of the neurones were heavily labelled in a Golgi-like manner, while others showed only weak labelling. eNOS immunopositivity was found in the blood vessels and in neurones. eNOS positive neurones were much more numerous than nNOS-containing cells, and represent about 60% of the cortical cells. However, with antibodies against eNOS, we never observed neurone-specific cell features. The NOS-containing cells found in our present study represent a possible morphological substrate for production of nitric oxide (NO).

大鼠视觉皮层一氧化氮合酶的免疫组织化学研究
本研究的目的是确定一氧化氮合酶(NOS)的两种亚型,神经元型(nNOS)和内皮型(eNOS)在大鼠视觉皮层中的分布。通过三种组织处理方案研究了每个NOS亚型的免疫组织化学定位。在第一个实验中,在30 μm厚的切片上用膜清洁剂(Triton或Saponin)进行免疫组化反应,以增加组织对抗体的通透性。在第二个方案中,我们将这些洗涤剂从所有溶液中排除,以避免破坏细胞膜。在第三个方案中,我们使用薄石蜡切片(5 μm厚)来确保抗体递送到细胞内结构。我们的数据表明,NOS的神经元和内皮亚型都存在于视觉皮层。在eNOS或nNOS抗体标记的神经元中,肯定存在一些兴奋性细胞。nNOS免疫阳性的是神经元和密集的纤维网络,可能是轴突。一些神经元以类似高尔基的方式被大量标记,而另一些神经元只显示出微弱的标记。eNOS免疫阳性见于血管和神经元。eNOS阳性神经元数量远多于含有nnos的细胞,约占皮层细胞的60%。然而,对于eNOS抗体,我们从未观察到神经元特异性细胞特征。在我们目前的研究中发现的含有一氧化氮的细胞代表了一种可能的形态基质,用于生产一氧化氮(NO)。
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