Chicken primordial germ cell motility in response to stem cell factor sensing.

Thanida Srihawong, T. Kuwana, K. Siripattarapravat, C. Tirawattanawanich
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引用次数: 11

Abstract

Avian primordial germ cells (PGCs) are destined to migrate a long distance from their extra embryonic region via the vascular system to the gonadal ridges where they form the germ cells. Although PGC migration is crucial for a genetic continuation to the next generation, the factors and mechanisms that control their migration remain largely unknown. In the present study the chemotactic effect of stem cell factor (SCF) was examined on chicken blood circulating PGCs (cPGC), employing 3D chemotaxis slides and time-lapsed imaging analyses as an in vitro study model. Upon in vitro exposure to an SCF gradient, 77.1% (54 out of 70) of cPGCs showed a clear response, of which 48.1% (26 out of 54) polarized with the consecutive formation of a persistent membrane protrusion and significant directional migration towards the gradient and the others showed transient membrane protrusions. In contrast, the controls and apparently SCF unresponsive cPGCs and c-kit-negative red blood cells (RBCs) showed only cytoplasmic cycling with random formations of membrane blebbing and no directional migration. Significant (p < 0.05) differences between the SCF-treated and control cPGCs and RBCs were found in the migration parameters of eccentricity, accumulated and Euclidean distances, and migration velocity. The SCF-treated PGCs also revealed a chemotactic response, as judged by their significant displacement of center of mass and Rayleigh test. Complete inhibition of all the SCF-induced responses in PGCs was found following pretreatment of the cPGCs with 10 µM of the c-kit inhibitor, STI57l, prior to SCF exposure. In addition, cPGCs were found to be positive for c-kit expression using a polyclonal goat anti-mouse c-kit primary antibody, suggesting that the cPGCs were capable of SCF sensing and the potential involvement of SCF/c-kit in the chemotactic migration. Therefore, SCF is suggested to function as a chemoattractant in the migration of chicken cPGC.
干细胞因子感应对鸡原始生殖细胞运动的影响。
鸟类原始生殖细胞(PGCs)注定要通过血管系统从胚胎外区域迁移到性腺脊,在那里它们形成生殖细胞。尽管PGC迁移对于遗传延续到下一代至关重要,但控制其迁移的因素和机制在很大程度上仍然未知。本研究采用三维趋化载玻片和延时成像分析作为体外研究模型,探讨了干细胞因子(SCF)对鸡血液循环PGCs (cPGC)的趋化作用。在体外暴露于SCF梯度时,77.1%(54 / 70)的cpgc表现出明显的反应,其中48.1%(26 / 54)极化,连续形成持久的膜突出,并向梯度方向明显迁移,其他cpgc表现出短暂的膜突出。相比之下,对照组和明显无SCF反应的cpgc和c-kit阴性红细胞(rbc)仅显示细胞质循环,随机形成膜泡,无定向迁移。scf处理的cpgc和红细胞在偏心率、累积距离和欧氏距离、迁移速度等参数上与对照组有显著差异(p < 0.05)。经scf处理的PGCs也显示出趋化反应,通过质心的显著位移和瑞利试验来判断。在暴露于SCF之前,用10µM的c-kit抑制剂STI57l预处理cpgc后,发现PGCs中所有SCF诱导的反应都被完全抑制。此外,使用山羊抗小鼠c-kit一抗发现cPGCs的c-kit表达阳性,这表明cPGCs能够感知SCF,并且SCF/c-kit可能参与趋化迁移。因此,SCF可能是鸡cPGC迁移的化学引诱剂。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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