GAD67-GFP基因敲入小鼠GFP与nNOS在嗅球中的表达和共存研究

GAD67-GFP expression and co-localization with bNOS in main olfactory bulb of GAD67-GFP knock-in mouse

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。

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.