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浙江大学学报（医学版）
浙江大学学报（医学版）  2012, Vol. 41 Issue (2): 159-165    DOI: 10.3785/j.issn.1008-9292.2012.02.006
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GAD67-GFP基因敲入小鼠GFP与nNOS在嗅球中的表达和共存研究
韩飞1，杨静1，余文富2，尹一飞1，武胜昔3，凌树才1
1．浙江大学医学院人体解剖与细胞生物学系，浙江 杭州 310058； 2．衢州职业技术学院医学院，浙江 衢州 324000； 3．第四军医大学人体解剖与组织胚胎学系，陕西 西安 710032
GAD67-GFP expression and co-localization with bNOS in main olfactory bulb of GAD67-GFP knock-in mouse
HAN Fei1,YANG Jing1,YU Wen-fu2,YIN Yi-fei1,WU Sheng-xi3,LING Shu-cai1
1.Department of Anatomy and Cell Biology,Zhejiang University School of Medicine,Hangzhou 310058,China; 2.College of Medicine,Quzhou College of Technology,Quzhou 324000,China; 3.Department of Anatomy and Histology,The Forth Military Medical University,Xi′an 710032,China
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摘要： 目的：探究GAD67-GFP基因敲入小鼠嗅球各细胞层中GAD67阳性神经元的分布及与一氧化氮合酶（bNOS）的共存。
方法：对该实验室繁育的成年GAD67-GFP基因敲入小鼠利用聚合酶链式反应(PCR)进行基因型等鉴定，取阳性GAD67-GFP基因敲入小鼠嗅球做冰冻切片进行尼氏染色、免疫荧光染色，观察GAD67阳性神经元在嗅球各细胞层中的分布比例及与bNOS共存。
结果：GAD67阳性神经元在嗅小球层、僧帽细胞层、颗粒细胞层所占比例分别为：（42.98±0.92）%、（23.64±0.84）%、（77.75±0.84）%，bNOS阳性神经元在球旁细胞层、僧帽细胞层有强阳性分布，而在颗粒细胞层几乎没有分布。GAD67与bNOS仅在嗅小球层偶有共存。
结论：GAD67阳性神经元主要分布于嗅小球层与颗粒细胞层， GAD67与bNOS仅少数共存于嗅小球层。
关键词： 谷氨酸脱羧酶/遗传学； 发光蛋白质类/遗传学； 嗅球/解剖学和组织学； 小鼠;  基因敲除； 神经元； 一氧化氮合酶； GAD67-GFP； GAD67； bNOS； 共存    
Abstract: Objective： To investigate the distribution of GAD67 and the co-localization with bNOS in the main olfactory bulb of GAD67-GFP knock-in mouse.
Methods： Polymerase chain reaction was applied to identify the genotype of GAD67-GFP knock-in mouse,the animals were sacrificed and frozen sections of olfactory bulb were prepared.The Nissl-staining was performed to show an framework of the neuron in the olfactory bulb.The distribution of GAD67 and co-localization with bNOS were detected by immunofluorescence technique.
Results： The proportion of GAD67-positive cells among DAPI-positive cells were (42.98±0.92)% in glomerular layer,(23.64±0.84)% in mitral cell layer and (77.75±0.84)% in granule cell layer; the bNOS-positive cells mainly existed in glomerular layer and mitral cell layer,very few in granule cell layer.No co-localization of GAD67 and bNOS in granule cell layer and mitral cell layer was found,but there was dispersed distribution in glomerular layer.
Conclusions： GAD67-positive neurons mainly appear in glomerular layer and granule cell layer,and the bNOS is mostly expressed in glomerular layer and mitral cell layer; while the co-localization of GAD67 and bNOS only occurs in glomerular layer of olfactory bulb.
Key words: Glutamate decarboxylase/genet    Luminescent proteins/genet    Olfactory bulb/anat    Mice,knockout    Neurons    Nitric oxide synthase    GAD67-GFP    GFP    bNOS
出版日期: 2012-03-25
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引用本文:

韩飞;杨静;余文富;尹一飞;武胜昔;凌树才. GAD67-GFP基因敲入小鼠GFP与nNOS在嗅球中的表达和共存研究[J]. 浙江大学学报（医学版）, 2012, 41(2): 159-165.

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https://www.zjujournals.com/med/CN/Y2012/V41/I2/159

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