Parobchak Nataliya, Rao Shivani, Negron Ariel, Schaefer Jennifer, Bhattacharya Moshmi, Radovick Sally, Babwah Andy V
Laboratory of Human Growth and Reproductive Development, Department of Pediatrics, Robert Wood Johnson Medical School, Rutgers, The State University of New Jersey, New Brunswick, New Jersey; School of Graduate Studies, Joint Graduate Program in Toxicology, Rutgers, The State University of New Jersey, Piscataway, New Jersey.
Laboratory of Human Growth and Reproductive Development, Department of Pediatrics, Robert Wood Johnson Medical School, Rutgers, The State University of New Jersey, New Brunswick, New Jersey; Department of Medicine, Robert Wood Johnson Medical School, Rutgers, The State University of New Jersey, New Brunswick, New Jersey.
F S Sci. 2020 Aug;1(1):67-77. doi: 10.1016/j.xfss.2020.06.001. Epub 2020 Jun 20.
To characterize the expression and signaling of uterine GPR83 in vivo in the nonpregnant and pregnant mouse and in vitro in human endometrial and nonendometrial cells.
Controlled laboratory study.
Not applicable.
Not applicable.
None.
Expression of uterine Gpr83 was determined by quantitative polymerase chain reaction throughout the estrous cycle and during early pregnancy in ovarian-stimulated and non-ovarian-stimulated mice and pregnant and pseudopregnant mice. Expression was also determined in ovariectomized mice after the administration of oil, E2, P4, or E2 + P4 and in stromal cells following 6 days of in vitro decidualization. GPR83 signaling was studied in human endometrial and embryonic kidney cell lines. Cells were treated by PEN, a GPR83 ligand, and PEN-induced extracellular signal-regulated kinase (ERK) phosphorylation was assayed under conditions that blocked Gα and/or β-arrestin signaling.
Uterine Gpr83 is expressed throughout the estrous cycle and during early pregnancy; expression increases dramatically at the time of uterine receptivity, embryo implantation, and stromal cell decidualization. In the ovariectomized mouse, hormone add-back reveals that Gpr83 expression is highly responsive to the combined treatment of E2 and P4, and studies in the ovarian-stimulated mouse show that expression is also very sensitive to changes in E2 and P4 and is therefore tightly regulated by E2 and P4. At the implantation site, expression is elevated up to D6 of pregnancy and then declines rapidly on D7 and D8, suggesting that if there is any involvement in decidualization, it is likely associated with primary but not secondary stromal cell decidualization. This premise was supported by the observation that stromal cell decidualization in vitro progresses with a decline in Gpr83 expression. In ERα/PR-expressing endometrial Ishikawa cells, GPR83 mediates PEN signals in a Gα-dependent manner, and studies conducted in HEK 293 cells lacking β-arrestin revealed that GPR83 also signals via a β-arrestin-dependent manner. When signaling by either one or both pathways is downregulated, cells exhibit a major reduction in responsiveness to PEN treatment, demonstrating that signaling by both pathways is significant.
We hypothesize that PEN/GPR83 signaling regulates uterine receptivity, embryo implantation, and primary stromal cell decidualization by coupling to Gα- and β-arrestin-dependent pathways.
在非妊娠和妊娠小鼠体内以及在人子宫内膜细胞和非子宫内膜细胞体外，对子宫GPR83的表达和信号传导进行表征。
对照实验室研究。
不适用。
不适用。
无。
通过定量聚合酶链反应测定在动情周期、卵巢刺激和未刺激的小鼠早期妊娠期间、妊娠和假孕小鼠中子宫Gpr83的表达。还测定了在给予油、E2、P4或E2 + P4后去卵巢小鼠中的表达以及体外蜕膜化6天后基质细胞中的表达。在人子宫内膜和胚胎肾细胞系中研究GPR83信号传导。用GPR83配体PEN处理细胞，并在阻断Gα和/或β - 抑制蛋白信号传导的条件下测定PEN诱导的细胞外信号调节激酶（ERK）磷酸化。
子宫Gpr83在整个动情周期和妊娠早期均有表达；在子宫接受性、胚胎着床和基质细胞蜕膜化时表达显著增加。在去卵巢小鼠中，激素补充显示Gpr83表达对E2和P4联合治疗高度敏感，在卵巢刺激小鼠中的研究表明表达对E2和P4的变化也非常敏感，因此受E2和P4严格调控。在着床部位，表达在妊娠第6天之前升高，然后在第7天和第8天迅速下降，这表明如果其参与蜕膜化，可能与初级而非次级基质细胞蜕膜化有关。体外基质细胞蜕膜化过程中Gpr83表达下降的观察结果支持了这一前提。在表达ERα/PR的子宫内膜 Ishikawa细胞中，GPR83以Gα依赖的方式介导PEN信号，在缺乏β - 抑制蛋白的HEK 293细胞中进行的研究表明GPR83也通过β - 抑制蛋白依赖的方式发出信号。当任一或两条途径的信号传导下调时，细胞对PEN处理的反应性大幅降低，表明两条途径的信号传导均很重要。
我们假设PEN/GPR83信号传导通过与Gα和β - 抑制蛋白依赖的途径偶联来调节子宫接受性、胚胎着床和初级基质细胞蜕膜化。
