Green fluorescent protein expression and colocalization with calretinin, parvalbumin, and somatostatin in the GAD67-GFP knock-in mouse
Green fluorescent protein expression and colocalization with calretinin, parvalbumin, and somatostatin in the GAD67-GFP knock-in mouse
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DOI：
10.1002/cne.10905
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发表时间：
2003-12-01
影响因子：
2.5
通讯作者：
Kaneko, T
中科院分区：
文献类型：
--
作者：
Tamamaki, N;Yanagawa, Y;Kaneko, T
中枢神经系统中的GABA能神经元调节其他神经元的活动，在信息处理中起着至关重要的作用。为了促进对GABA能神经元的研究，我们制备了两株谷氨酸脱羧酶-绿色荧光蛋白(GAD67-GFP)敲入小鼠。GFP阳性胞体的分布模式与GAD67原位杂交信号在中枢神经系统的分布相同。我们既没有在GAD67阴性细胞中发现明显的异位GFP表达，也没有在两个GAD67-GFP敲入小鼠系中GAD67阳性神经元中明显缺乏GFP表达。GFP的表达时间与GAD67的表达时间也是平行的。因此，我们构建了一张GFP在敲入小鼠大脑中的分布图。此外，我们使用敲入小鼠来研究GFP与NeuN、钙维A(CR)、小白蛋白(PV)和生长抑素(SS)在额叶运动皮质中的共定位。GFP阳性细胞在NeuN阳性细胞(新皮质神经元)中的比例约为19.5%。所有CR、PV和SS阳性细胞均呈GFP阳性。CR、PV和SS阳性细胞发出不同强度的GFP荧光。CR、PV和SS阳性细胞占GFP阳性细胞的比例分别为13.9%、40.1%和23.4%。因此，这三种类型的GABA能神经元占GFP阳性细胞的77.4%。与未鉴定的GFP阳性细胞一致，在第I层观察到许多发出强烈GFP荧光的中等大小的球形体细胞。(C)2003 Wiley-Liss，Inc.
gamma-aminobutyric acid (GABA)ergic neurons in the central nervous system regulate the activity of other neurons and play a crucial role in information processing. To assist an advance in the research of GABAergic neurons, here we produced two lines of glutamic acid decarboxylase-green fluorescence protein (GAD67-GFP) knock-in mouse. The distribution pattern of GFP-positive somata was the same as that of the GAD67 in situ hybridization signal in the central nervous system. We encountered neither any apparent ectopic GFP expression in GAD67-negative cells nor any apparent lack of GFP expression in GAD67-positive neurons in the two GAD67-GFP knock-in mouse lines. The timing of GFP expression also paralleled that of GAD67 expression. Hence, we constructed a map of GFP distribution in the knock-in mouse brain. Moreover, we used the knock-in mice to investigate the colocalization of GFP with NeuN, calretinin (CR), parvalbumin (PV), and somatostatin (SS) in the frontal motor cortex. The proportion of GFP-positive cells among NeuN-positive cells (neocortical neurons) was approximately 19.5%. All the CR, PV, and SS-positive cells appeared positive for GFP. The CR-, PV, and SS-positive cells emitted GFP fluorescence at various intensities characteristics to them. The proportions of CR-, PV-, and SS-positive cells among GFP-positive cells were 13.9%, 40.1%, and 23.4%, respectively. Thus, the three subtypes of GABAergic neurons accounted for 77.4% of the GFP-positive cells. They accounted for 6.5% in layer I. In accord with unidentified GFP-positive cells, many medium-sized spherical somata emitting intense GFP fluorescence were observed in layer I. (C) 2003 Wiley-Liss, Inc.