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不同哺乳动物(包括人类)脑组织中双皮质素和 Ki67 抗原检测的一致性和变异性。

Consistency and Variation in Doublecortin and Ki67 Antigen Detection in the Brain Tissue of Different Mammals, including Humans.

机构信息

Neuroscience Institute Cavalieri Ottolenghi (NICO), 10043 Orbassano, Italy.

Department of Veterinary Sciences, University of Turin, 10095 Torino, Italy.

出版信息

Int J Mol Sci. 2023 Jan 28;24(3):2514. doi: 10.3390/ijms24032514.

Abstract

Recently, a population of "immature" neurons generated prenatally, retaining immaturity for long periods and finally integrating in adult circuits has been described in the cerebral cortex. Moreover, comparative studies revealed differences in occurrence/rate of different forms of neurogenic plasticity across mammals, the "immature" neurons prevailing in gyrencephalic species. To extend experimentation from laboratory mice to large-brained mammals, including humans, it is important to detect cell markers of neurogenic plasticity in brain tissues obtained from different procedures (e.g., post-mortem/intraoperative specimens vs. intracardiac perfusion). This variability overlaps with species-specific differences in antigen distribution or antibody species specificity, making it difficult for proper comparison. In this work, we detect the presence of doublecortin and Ki67 antigen, markers for neuronal immaturity and cell division, in six mammals characterized by widely different brain size. We tested seven commercial antibodies in four selected brain regions known to host immature neurons (paleocortex, neocortex) and newly born neurons (hippocampus, subventricular zone). In selected human brains, we confirmed the specificity of DCX antibody by performing co-staining with fluorescent probe for DCX mRNA. Our results indicate that, in spite of various types of fixations, most differences were due to the use of different antibodies and the existence of real interspecies variation.

摘要

最近,在大脑皮层中描述了一群在产前生成的“不成熟”神经元,它们保持长时间的不成熟状态,最终整合到成年回路中。此外,比较研究显示,不同形式的神经发生可塑性在哺乳动物中的发生/速率存在差异,“不成熟”神经元在脑回较多的物种中更为普遍。为了将实验室小鼠的实验扩展到大脑较大的哺乳动物,包括人类,检测不同程序(例如,死后/手术标本与心脏内灌注)获得的脑组织中的神经发生可塑性的细胞标志物非常重要。这种可变性与抗原分布或抗体物种特异性的物种特异性差异重叠,使得难以进行适当的比较。在这项工作中,我们检测了六种具有广泛不同大脑大小的哺乳动物中双皮质素和 Ki67 抗原(神经元不成熟和细胞分裂的标志物)的存在。我们在四个已知存在不成熟神经元(古皮层、新皮层)和新生神经元(海马、室下区)的选定脑区测试了七种商业抗体。在选定的人类大脑中,我们通过与 DCX mRNA 的荧光探针进行共染色来确认 DCX 抗体的特异性。我们的结果表明,尽管存在各种固定类型,但大多数差异是由于使用了不同的抗体以及存在真正的种间变异。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4ba5/9916846/9b55b763277d/ijms-24-02514-g005a.jpg

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