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凝集素作为一种检测成年小鼠大脑中神经干细胞/祖细胞的工具。

Lectins as a tool for detecting neural stem/progenitor cells in the adult mouse brain.

机构信息

Department of Stem Cell Biology and Histology, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan.

出版信息

Anat Rec (Hoboken). 2011 Feb;294(2):305-21. doi: 10.1002/ar.21311. Epub 2010 Dec 3.

Abstract

Glycoconjugates are biopolymers that are broadly distributed in the central nervous system, including the cell surface of neural stem cells or neural precursor cells (NSCs/NPCs). Glycoconjugates can be recognized by carbohydrate-binding proteins, lectins. Two lectins, Phaseolus vulgaris lectin agglutinin E-form (PHA-E4) and wheat germ agglutinin (WGA) have been reported to be useful in isolating NSCs/NPCs by fluorescence-activated cell sorting (FACS) or immunopanning methods. In this study, we analyzed the lectin-binding properties of NSCs/NPCs in two neurogenic regions of the adult mouse brain to determine whether PHA-E4 and WGA exhibit specific binding patterns on sections and whether there are other lectins presenting the binding pattern similar to those of PHA-E4 and WGA in lectin histochemistry. Among nine types of lectins, peanut agglutinin was localized to the white matter and four lectins bound to cells within the subventricular zone (SVZ) of the lateral ventricle. Lectin histochemistry combined with immunohistochemistry demonstrated that one lectin, Ricinus communis agglutinin, specifically detected type A neuronal precursors and that the remaining three lectins, Agaricus bisporus agglutinin (ABA), PHA-E4, and WGA, recognized type B NSCs and type C transient amplifying cells in the SVZ. These three lectins also recognized type 1 quiescent neural progenitors and type 2a amplifying neural progenitors in the subgranular layer of the dentate gyrus. Lectin histochemistry of the neurosphere culture also yielded similar results. These observations suggest that, in addition to PHA-E4 and WGA, ABA lectin may also be applicable in FACS or immunopanning for the isolation of NSCs/NPCs.

摘要

糖缀合物广泛分布于中枢神经系统,包括神经干细胞或神经前体细胞(NSCs/NPCs)的细胞表面。糖缀合物可以被糖结合蛋白(凝集素)识别。两种凝集素,菜豆植物凝集素 E 型(PHA-E4)和麦胚凝集素(WGA)已被报道可用于通过荧光激活细胞分选(FACS)或免疫淘选方法分离 NSCs/NPCs。在这项研究中,我们分析了成年小鼠大脑两个神经发生区域的 NSCs/NPC 的凝集素结合特性,以确定 PHA-E4 和 WGA 是否在切片上表现出特异性结合模式,以及是否存在其他凝集素在凝集素组织化学中呈现与 PHA-E4 和 WGA 相似的结合模式。在 9 种类型的凝集素中,花生凝集素定位于白质,4 种凝集素结合于侧脑室 SVZ 内的细胞。凝集素组织化学结合免疫组织化学显示,一种凝集素,蓖麻凝集素,特异性检测 A 型神经元前体,其余 3 种凝集素,姬松茸凝集素(ABA)、PHA-E4 和 WGA,识别 SVZ 中的 B 型 NSCs 和 C 型短暂扩增细胞。这 3 种凝集素也识别齿状回颗粒下层的 1 型静止神经祖细胞和 2a 型扩增神经祖细胞。神经球培养的凝集素组织化学也得到了类似的结果。这些观察结果表明,除了 PHA-E4 和 WGA 之外,ABA 凝集素也可能适用于 FACS 或免疫淘选以分离 NSCs/NPCs。

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