Division of General Pediatrics, Department of Pediatrics and Adolescent Medicine, Medical University of Graz, 8036 Graz, Austria.
Division of Pediatric Hematology and Oncology, Department of Pediatrics and Adolescent Medicine, Faculty of Medicine, Medical Center-University of Freiburg, 79098 Freiburg im Breisgau, Germany.
Cells. 2022 Aug 17;11(16):2563. doi: 10.3390/cells11162563.
Despite fully functional primary hemostasis, platelets of healthy neonates exhibit hypoaggregability and secretion defects, which may be adaptations to specific requirements in this developmental stage. The etiologies for reduced signal transduction vary with the type of agonist. The discovered peculiarities are lower receptor densities, reduced calcium mobilization, and functional impairments of G proteins. Reduced secretion of dense granules has been attributed to lower numbers of granules. Signaling studies with adult platelets have shown a regulating effect of the G signaling pathway on dense granule secretion via RhoA. We comparatively analyzed secretion profiles using flow cytometry and expression levels of G, G, and G using Western blot analysis in platelets from cord blood and adults. Furthermore, we evaluated Rho activation after in vitro platelet stimulation with thrombin using a pulldown assay. We observed a markedly reduced expression of the dense granule marker CD63 on neonatal platelets after thrombin stimulation. G expression was significantly decreased in neonatal platelets and correlated with lower Rho activation after thrombin stimulation. We conclude that lower expression of G in neonatal platelets results in attenuated activation of Rho and may contribute to reduced secretion of dense granules after exposure to thrombin.
尽管新生儿的初级止血功能正常,但血小板表现出低聚集性和分泌缺陷,这可能是对该发育阶段特定需求的适应。信号转导减少的病因因激动剂的类型而异。已发现的特点是受体密度降低、钙动员减少以及 G 蛋白功能障碍。致密颗粒分泌减少归因于颗粒数量减少。使用成人血小板进行的信号转导研究表明,G 信号通路通过 RhoA 对致密颗粒分泌具有调节作用。我们使用流式细胞术比较分析了来自脐血和成人血小板的分泌谱,并使用 Western blot 分析了 G、G 和 G 的表达水平。此外,我们还使用下拉测定法评估了体外血小板用凝血酶刺激后的 Rho 激活。我们观察到,凝血酶刺激后新生儿血小板上致密颗粒标记物 CD63 的表达明显降低。新生儿血小板中的 G 表达显著降低,与凝血酶刺激后 Rho 激活降低相关。我们得出结论,新生儿血小板中 G 的表达降低导致 Rho 的激活减弱,可能导致暴露于凝血酶后致密颗粒分泌减少。