Endogenous biotin as a marker of adrenocortical cells with steroidogenic potential.
Endogenous biotin as a marker of adrenocortical cells with steroidogenic potential.
复制标题
内源性生物素作为具有类固醇生成潜力的肾上腺皮质细胞的标记物。
DOI：
10.1016/j.mce.2011.01.015
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发表时间：
2011
影响因子：
4.1
通讯作者：
Laufer,Ed
中科院分区：
文献类型：
--
作者：
Paul,Alex;Laufer,Ed
通过在单细胞分辨率下同时检查多个标记物，可以极大地促进肾上腺皮质表型的解释。然而，多种合适抗体的可用性可能是限速的，而它们的同源抗原往往是可变的可及性。因此，不受这些约束的特定标记具有明显的实用性。在这里，我们报告了内源性生物素，当使用荧光链亲和素在固定的冷冻组织切片中检测时，是小鼠肾上腺皮质中具有类固醇生成潜力的所有细胞的特异性标记物。虽然链霉亲和素可以染色体甾原性和成熟皮质细胞，但它不能标记肾上腺被膜、髓质或血管内皮。发育档案显示从E13.5到成年期肾上腺内源性生物素标记。与区域标记、下丘脑-垂体-肾上腺（HPA）轴重塑组织、转基因sh - nlacz或Gli1-nLacZ动物和Shh突变胚胎的比较进一步证明了该方法的实用性。荧光链亲和素应用于一个简单的一步染色方案，因此提供了一个有效的反染色用于肾上腺分析。
Interpretation of adrenal cortex phenotypes is greatly facilitated by simultaneous examination of multiple markers at single cell resolution. However, the availability of multiple appropriate antibodies can be rate limiting, while their cognate antigens are often subject to variable accessibility. Specific markers not subject to these constraints thus have obvious utility. Here we report that endogenous biotin, when detected in fixed, frozen tissue sections using fluorescent streptavidin, is a specific marker of apparently all cells with steroidogenic potential in the murine adrenal cortex. While streptavidin stains presteroidogenic and mature cortical cells, it does not label the adrenal capsule, medulla or vascular endothelium. Developmental profiles reveal adrenal endogenous biotin labeling from E13.5 through adulthood. Comparisons with zonal markers, hypothalamic–pituitary–adrenal (HPA) axis-remodeled tissue, transgenic Shh-nLacZ or Gli1-nLacZ animals, and Shh mutant embryos further demonstrate the utility of this approach. Fluorescent streptavidin applied using a simple one-step staining protocol thus provides a potent counterstain for use in adrenal analyses.