Regulation of Kit Expression in Early Mouse Embryos and ES Cells.
Regulation of Kit Expression in Early Mouse Embryos and ES Cells.
复制标题
DOI：
10.1002/stem.2960
复制
发表时间：
2019-03
期刊：
影响因子：
--
通讯作者：
Dolci S
中科院分区：
文献类型：
--
作者：
Todaro F;Campolo F;Barrios F;Pellegrini M;Di Cesare S;Tessarollo L;Rossi P;Jannini EA;Dolci S
Kit是一种生长因子受体，调节许多胚胎和出生后干细胞类型的增殖和/或存活。当突变时，它可以诱导宿主细胞的恶性转化。为了剖析Kit在ESC多能性控制中的作用，我们研究了其在早期小鼠胚胎发生过程中的表达以及在ESC从内细胞团（ICM）细胞衍生的过程中的表达。我们跟踪了从携带与EGFP融合的Kit启动子区域（Kit-EGFP）的转基因小鼠获得的早期小鼠胚胎的体外发育，发现它们在桑葚胚阶段启动EGFP表达。然后EGFP表达维持在胚泡中的ICM内，并且与仅LIF条件相比，当在MAPK和GSK 3 β抑制剂（2 i）加LIF存在下培养时，其水平增加。当在LIF条件下培养时，Kit-EGFP ESCs显示不均一的EGFP表达模式，但当转移到2 i-LIF培养时，它们上调EGFP表达，以及Sox 2、Nanog、Prdm 14的表达。类似地，原始生殖细胞（PGCs）在胚胎生殖细胞（EGC）转化过程中在2 i-LIF中显示增强的EGFP表达。试剂盒的表达受到操纵Sox 2水平在胚胎干细胞。染色质免疫沉淀实验证实，Sox 2结合试剂盒调控区含有Sox 2的共识序列。最后，由D814 Y突变诱导的Kit组成型激活增加了体外和体内畸胎瘤测定中的ESC增殖和克隆效率。我们的研究结果确定Kit作为一个多能性反应基因，并建议在ESC增殖的调节Kit的作用。
Kit is a growth factor receptor that regulates proliferation and/or survival of many embryonic and postnatal stem cell types. When mutated, it can induce malignant transformation of the host cells. To dissect the Kit role in the control of ESC pluripotency, we studied its expression during early mouse embryogenesis and during the process of ESC derivation from inner cell mass (ICM) cells. We followed the in vitro development of early mouse embryos obtained from transgenic mice carrying Kit promoter regions fused to EGFP (Kit-EGFP) and found that they initiate EGFP expression at morula stage. EGFP expression is then maintained in the blastocyst, within the ICM, and its levels increase when cultured in the presence of MAPK and GSK3β inhibitors (2i) plus LIF compared with the LIF-only condition. Kit-EGFP ESCs showed nonhomogeneous EGFP expression pattern when cultured in LIF condition, but they upregulated EGFP expression, as well as that of Sox2, Nanog, Prdm14, when shifted to 2i-LIF culture. Similarly, primordial germ cells (PGCs) in the process of embryonic germ cell (EGC) conversion showed enhanced EGFP expression in 2i-LIF. Kit expression was affected by manipulating Sox2 levels in ESCs. Chromatin immunoprecipitation experiments confirmed that Sox2 binds Kit regulatory regions containing Sox2 consensus sequences. Finally, Kit constitutive activation induced by the D814Y mutation increased ESC proliferation and cloning efficiency in vitro and in teratoma assays in vivo. Our results identify Kit as a pluripotency-responsive gene and suggest a role for Kit in the regulation of ESC proliferation.
登录
查看更多内容
影响因子：
9
作者：
Di Siena S;Gimmelli R;Nori SL;Barbagallo F;Campolo F;Dolci S;Rossi P;Venneri MA;Giannetta E;Gianfrilli D;Feigenbaum L;Lenzi A;Naro F;Cianflone E;Mancuso T;Torella D;Isidori AM;Pellegrini M
通讯作者：
Pellegrini M
影响因子：
2.7
作者：
Farini, D;Scaldaferri, ML;De Felici, M
通讯作者：
De Felici, M
影响因子：
23.9
作者：
Hayashi K;de Sousa Lopes SMC;Tang F;Lao K;Surani MA
通讯作者：
Surani MA
影响因子：
25
作者：
Favaro, Rebecca;Valotta, Menella;Nicolis, Silvia K.
通讯作者：
Nicolis, Silvia K.
影响因子：
64.5
作者：
Marson A;Levine SS;Cole MF;Frampton GM;Brambrink T;Johnstone S;Guenther MG;Johnston WK;Wernig M;Newman J;Calabrese JM;Dennis LM;Volkert TL;Gupta S;Love J;Hannett N;Sharp PA;Bartel DP;Jaenisch R;Young RA
通讯作者：
Young RA