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成年灵长类动物大脑皮层中不存在双皮质素(DCX)阳性神经发生。

No DCX-positive neurogenesis in the cerebral cortex of the adult primate.

作者信息

Liu Ruo-Xu, Ma Jie, Wang Bin, Tian Tian, Guo Ning, Liu Shao-Jun

机构信息

Institute of Military Cognition and Brain Sciences, Beijing, China.

Department of Pharmacy, Medical College, Huanghe S&T University, Zhengzhou, Henan Province, China.

出版信息

Neural Regen Res. 2020 Jul;15(7):1290-1299. doi: 10.4103/1673-5374.272610.

Abstract

Whether endogenous neurogenesis occurs in the adult cortex remains controversial. An increasing number of reports suggest that doublecortin (DCX)-positive neurogenesis persists in the adult primate cortex, attracting enormous attention worldwide. In this study, different DCX antibodies were used together with NeuN antibodies in immunohistochemistry and western blot assays using adjacent cortical sections from adult monkeys. Antibody adsorption, antigen binding, primary antibody omission and antibody-free experiments were used to assess specificity of the signals. We found either strong fluorescent signals, medium-weak intensity signals in some cells, weak signals in a few perikarya or near complete lack of labeling in adjacent cortical sections incubated with the various DCX antibodies. The putative DCX-positive cells in the cortex were also positive for NeuN, a specific marker of mature neurons. However, further experiments showed that most of these signals were either the result of antibody cross reactivity, the non-specificity of secondary antibodies or tissue autofluorescence. No confirmed DCX-positive cells were detected in the adult macaque cortex by immunofluorescence. Our findings show that DCX-positive neurogenesis does not occur in the cerebral cortex of adult primates, and that false-positive signals (artefacts) are caused by antibody cross reactivity and autofluorescence. The experimental protocols were approved by the Institutional Animal Care and Use Committee of the Institute of Neuroscience, Beijing, China (approval No. IACUC-AMMS-2014-501).

摘要

内源性神经发生是否发生在成年皮质中仍存在争议。越来越多的报告表明,双皮质素(DCX)阳性的神经发生在成年灵长类动物皮质中持续存在,这在全球引起了极大关注。在本研究中,在免疫组织化学和蛋白质印迹分析中,将不同的DCX抗体与NeuN抗体一起用于成年猴子相邻皮质切片。采用抗体吸附、抗原结合、省略一抗和无抗体实验来评估信号的特异性。我们发现,在用各种DCX抗体孵育的相邻皮质切片中,要么有强荧光信号,一些细胞中有中弱强度信号,少数核周体中有弱信号,要么几乎完全没有标记。皮质中假定的DCX阳性细胞对NeuN(成熟神经元的特异性标志物)也呈阳性。然而,进一步的实验表明,这些信号大多是抗体交叉反应、二抗非特异性或组织自发荧光的结果。通过免疫荧光在成年猕猴皮质中未检测到确认的DCX阳性细胞。我们的研究结果表明,成年灵长类动物的大脑皮质中不会发生DCX阳性的神经发生,并且假阳性信号(假象)是由抗体交叉反应和自发荧光引起的。实验方案已获得中国北京神经科学研究所实验动物管理与使用委员会的批准(批准号:IACUC-AMMS-2014-501)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5794/7047795/57d5b96cd746/NRR-15-1290-g002.jpg

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