Lüth H J, Hedlich A, Hilbig H, Winkelmann E, Mayer B
University of Leipzig, Paul-Flechsig-Institute of Brain Research, Department of Neuroanatomy.
J Hirnforsch. 1995;36(3):313-28.
The postnatal development of NADPH-diaphorase (NADPH-d)/nitric oxide synthase (NOS) positive nerve cells was studied in the visual cortex of rats on postnatal day 1, 5, 10, 15, 20, 30 and at the age of 1 year. NADPH-d was demonstrated enzymhistochemically and NOS immunohistochemically using a polyclonal antibody. NADPH-d is localized in nerve cell somata, dendrites, axons and blood vessels, whereas NOS immunoreactivity is only detectable in nerve cells. The identity of NADPH-d cells with those which contain NOS was proved in double labelling experiments in the cortex of rats on postnatal day 5, 15 and at the age of 1 year. The results of these experiments have shown that in the cortex of rats NADPH-d positive cells are identical with NOS-positive cells in the different stages. Therefore we have used NADPH-d histochemistry in all other postnatal stages as a marker for neurons which contain NOS. NOS positive nerve cells appear very early on postnatal day 1 in the intermediate (white matter) and subplate (layers V and VI) region as small undifferentiated neurons. During the following postnatal differentiation these neurons reached their typical morphology in the second week and appeared in all layers. Neurons in layers V and VI preceded those in the superficial layers. Nerve cells in the white matter seem to have their own differentiation pattern because they showed characteristic features of immaturated varicose dendrites for a longer time. The investigation of soma size with the computerized "Kontron Videoplan" system (Zeiss, Germany) showed the largest cell bodies on postnatal day 20 which then decreased towards adulthood. Between postnatal day 10 and 20 some NOS-positive neurons especially in the deep layers displayed symptoms of degeneration, like shrunken cell bodies, corkscrew and twisted dendrites. Furthermore, NOS-positive neurons in layer I are not detectable in adult neocortex. These observations could suggest that some NOS-positive cells in the cerebral cortex of rats may occur only transiently. Also in the neuropil some alterations in the localization of NOS positive axonal boutons were observed. On postnatal day 10 NOS negative cell somata were shadowy surrounded by boutons. During the further development from postnatal day 20 until adulthood this particular position was no longer visible. Beside the NOS cells which played a transient role, the majority of these cells survived to adulthood and are a morphological (Martinotti-cells with ascending axons) and chemical (GABAergic, NADPH-d/NOS positive, peptide containing cells) defined cell type in the neuronal network of the cortex of the rat.
研究了出生后第1天、5天、10天、15天、20天、30天以及1岁大鼠视觉皮层中烟酰胺腺嘌呤二核苷酸磷酸黄递酶（NADPH-d）/一氧化氮合酶（NOS）阳性神经细胞的发育情况。采用酶组织化学方法显示NADPH-d，并用多克隆抗体进行NOS免疫组织化学检测。NADPH-d定位于神经细胞体、树突、轴突和血管，而NOS免疫反应性仅在神经细胞中可检测到。在出生后第5天、15天和1岁大鼠皮层的双重标记实验中，证实了NADPH-d阳性细胞与含NOS的细胞相同。这些实验结果表明，在大鼠皮层中，NADPH-d阳性细胞在不同阶段与NOS阳性细胞相同。因此，在所有其他出生后阶段我们都使用NADPH-d组织化学作为含NOS神经元的标记。出生后第1天，NOS阳性神经细胞最早出现在中间（白质）和板下层（V层和VI层）区域，为未分化的小神经元。在出生后的后续分化过程中，这些神经元在第二周达到其典型形态，并出现在所有层中。V层和VI层的神经元比表层的神经元出现得早。白质中的神经细胞似乎有其自身的分化模式，因为它们在较长时间内表现出未成熟曲张树突的特征。使用计算机化的“Kontron Videoplan”系统（德国蔡司公司）对细胞体大小进行的研究表明，出生后第天细胞体最大，随后向成年期逐渐减小。在出生后第10天至20天之间，一些NOS阳性神经元，尤其是深层的神经元，出现了退化症状，如细胞体萎缩、螺旋状和扭曲的树突。此外，在成年新皮层中未检测到I层中的NOS阳性神经元。这些观察结果可能表明，大鼠大脑皮层中的一些NOS阳性细胞可能只是短暂出现。在神经毡中也观察到了NOS阳性轴突终扣定位的一些变化。出生后第10天，NOS阴性细胞体被终扣模糊地包围。在从出生后第20天到成年的进一步发育过程中，这种特殊位置不再可见。除了起短暂作用的NOS细胞外，这些细胞中的大多数存活到成年，并且是大鼠皮层神经网络中一种形态学（具有上升轴突的Martinotti细胞）和化学（GABA能、NADPH-d/NOS阳性、含肽细胞）定义的细胞类型。
