Identification of Cell Autonomous and Non-Cell Autonomous Functions of Heparan Sulfate Glycosaminoglycan Chains by Creating Chimeric Mouse Embryos
Identification of Cell Autonomous and Non-Cell Autonomous Functions of Heparan Sulfate Glycosaminoglycan Chains by Creating Chimeric Mouse Embryos
复制标题
通过创建嵌合小鼠胚胎鉴定硫酸乙酰肝素糖胺聚糖链的细胞自主和非细胞自主功能
DOI：
10.1007/978-1-0716-1398-6_44
复制
发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Kimura-Yoshida C.
中科院分区：
文献类型：
--
作者：
Matsuo I;Kimura-Yoshida C.
细胞表面栓系的硫酸乙酰肝素糖胺聚糖链主要以细胞自主方式起作用，而细胞外基质相关的硫酸乙酰肝素糖胺聚糖链以非细胞自主方式起作用。此外，由蛋白酶和乙酰肝素酶酶促释放的细胞表面束缚的硫酸乙酰肝素链的裂解形式（称为脱落）可以促进非细胞自主机制。硫酸乙酰肝素链通过转胞吞作用或丝状伪足介导的向周围细胞的移动还涉及另一种非细胞自主机制。为了确定硫酸乙酰肝素糖胺聚糖链在早期胚胎发生中的细胞自主或非细胞自主作用，可以通过分析由野生型和硫酸乙酰肝素糖胺聚糖链缺陷细胞组成的嵌合胚胎得出直接结论。在这里，我们描述了这些嵌合体胚胎的生产方法，并在单细胞水平上用免疫组织化学对其细胞表型进行分析。
Cell surface-tethered heparan sulfate glycosaminoglycan chains primarily function in a cell autonomous manner, while extracellular matrix-associated heparan sulfate glycosaminoglycan chains function in a non-cell autonomous manner. In addition, the cleaved forms of cell surface-tethered heparan sulfate chains enzymatically released by proteases and heparanases, called shedding, can contribute to non-cell autonomous mechanisms. The movement of heparan sulfate chains to surrounding cells mediated by transcytosis or filopodia also involves another non-cell autonomous mechanism. To determine cell autonomous or non-cell autonomous roles of heparan sulfate glycosaminoglycan chains during early embryogenesis, direct conclusions can be drawn by analyzing chimeric embryos which are composed of wild-type and heparan sulfate glycosaminoglycan chain-deficient cells. Here, we describe methods of production of these chimeric embryos and analysis of their cellular phenotypes with immunohistochemistry at a single-cell level.