Parrish-Aungst S, Shipley M T, Erdelyi F, Szabo G, Puche A C
Department of Anatomy and Neurobiology, Program in Neuroscience, University of Maryland School of Medicine, Baltimore, Maryland 21201, USA.
J Comp Neurol. 2007 Apr 20;501(6):825-36. doi: 10.1002/cne.21205.
Olfactory sensory information is processed and integrated by circuits within the olfactory bulb. Golgi morphology suggests the olfactory bulb contains several major neuronal classes. However, an increasingly diverse collection of neurochemical markers have been localized in subpopulations of olfactory bulb neurons. While the mouse is becoming the animal model of choice for olfactory research, little is known about the proportions of neurons expressing and coexpressing different neurochemical markers in this species. Here we characterize neuronal populations in the mouse main olfactory bulb, focusing on glomerular populations. Immunofluorescent labeling for: 1) calretinin, 2) calbindin D-28K (CB), 3) parvalbumin, 4) neurocalcin, 5) tyrosine hydroxylase (TH), 6) the 67-kDa isoform of GAD (GAD67), and 7) the neuronal marker NeuN was performed in mice expressing green fluorescent protein under the control of the glutamic acid decarboxylase 65kDa (GAD65) promoter. Using unbiased stereological cell counts we estimated the total numbers of cells and neurons in the bulb and the number and percentage of neurons expressing and coexpressing different neurochemical populations in each layer of the olfactory bulb. Use of a genetic label for GAD65 and immunohistochemistry for GAD67 identified a much larger percentage of GABAergic neurons in the glomerular layer (55% of all neurons) than previously recognized. Additionally, while many glomerular neurons expressing TH or CB coexpress GAD, the majority of these neurons preferentially express the GAD67 isoform. These data suggest that the chemospecific populations of neurons in glomeruli form distinct subpopulations and that GAD isoforms are preferentially regulated in different neurochemical cell types.
嗅觉感觉信息由嗅球内的神经回路进行处理和整合。高尔基形态学表明嗅球包含几种主要的神经元类型。然而,越来越多不同种类的神经化学标志物已定位在嗅球神经元的亚群中。虽然小鼠正成为嗅觉研究的首选动物模型,但对于该物种中表达和共表达不同神经化学标志物的神经元比例知之甚少。在这里,我们对小鼠主嗅球中的神经元群体进行了表征,重点关注肾小球群体。对以下标志物进行了免疫荧光标记:1)钙视网膜蛋白,2)钙结合蛋白D-28K(CB),3)小白蛋白,4)神经钙蛋白,5)酪氨酸羟化酶(TH),6)67-kDa亚型的谷氨酸脱羧酶(GAD67),以及7)神经元标志物NeuN,实验对象为在谷氨酸脱羧酶65kDa(GAD65)启动子控制下表达绿色荧光蛋白的小鼠。我们使用无偏立体细胞计数法估计了嗅球中细胞和神经元的总数,以及嗅球各层中表达和共表达不同神经化学群体的神经元数量和百分比。使用GAD65的基因标记和GAD67的免疫组织化学方法发现,肾小球层中GABA能神经元的比例(占所有神经元的55%)比之前认为的要高得多。此外,虽然许多表达TH或CB的肾小球神经元共表达GAD,但这些神经元中的大多数优先表达GAD67亚型。这些数据表明,肾小球中具有化学特异性的神经元群体形成了不同的亚群,并且GAD亚型在不同的神经化学细胞类型中受到优先调控。