Three-dimensional distribution patterns of newborn neurons in the adult olfactory bulb
Three-dimensional distribution patterns of newborn neurons in the adult olfactory bulb
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DOI：
10.1016/j.jneumeth.2009.06.009
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发表时间：
2009-09-15
影响因子：
3
通讯作者：
Mizrahi, Adi
中科院分区：
文献类型：
--
作者：
Kopel, Hagit;Meshulam, Meir;Mizrahi, Adi
我们提出了一种新的方法来研究小鼠嗅球（OB）新生神经元的三维空间分布模式。新生的神经元，在体内转导，使用慢病毒表达绿色荧光蛋白（GFP）。双光子（2P）显微镜成像厚OB切片（类似于250 μ m）在单细胞分辨率。图像堆栈被半自动捕获，并离线连接，以创建包含所有标记神经元的位置信息的更大的图像堆栈。大图像堆栈的连续重建产生了三维虚拟模型，其中包含OB大体积内所有标记的新生神经元的确切位置。通过分析数千个GFP标记的新生神经元的细胞分布，证明了该方法的可行性。该分析确定了其中新生细胞的密度显著高于平均密度的3D簇。我们表明，我们的方法揭示了在2D中仅对一小部分组织进行采样时被忽视的信息。这种方法不仅可以作为分析OB中新生神经元的一种有价值的工具，而且还可以用于其他神经元类型以及其他脑区域。(C)2009 Elsevier B.V.保留所有权利。
We present a new method to study the three-dimensional (3D) spatial distribution patterns of newborn neurons in the mouse olfactory bulb (OB). Newborn neurons were transduced, in vivo, using lentiviruses to express green fluorescent protein (GFP).Two-photon (2P) microscopy was used to image thick OB slices (similar to 250 mu m) at single cell resolution. Image-stacks were captured semi-automatically, and concatenated offline, to create larger image-stacks containing the positional information of all the labeled neurons. Serial reconstruction of the large image-stacks resulted in a three-dimensional virtual model, containing the exact position of all the labeled newborn neurons within large volumes of the OB. The feasibility of this method was demonstrated by analyzing the cell distributions of thousands of GFP labeled newborn neurons. This analysis identified 3D clusters in which the newborn cells' density is significantly higher than the mean density. We show that our method reveals information that is overlooked when sampling only a small fraction of the tissue in 2D. This method may serve as a valuable tool, not only for analyzing newborn neurons in the OB, but also for other neuronal types as well as for other brain regions. (C) 2009 Elsevier B.V. All rights reserved.