Centre National de la Recherche Scientifique, Institut Français du Cheval et de l'Equitation, Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement, INRAE, Université de Tours, Physiologie de la Reproduction et des Comportements, UMR85, 37380, Nouzilly, France.
SYSAAF-Syndicat des Sélectionneurs Avicoles et Aquacoles Français, Centre INRA Val de Loire, 37380, Nouzilly, France.
Sci Rep. 2022 May 29;12(1):8989. doi: 10.1038/s41598-022-12961-4.
Understanding of the distribution of chemerin and its receptors, Chemokine-like Receptor 1 (CMKLR1), G Protein-coupled Receptor 1 (GPR1) and Chemokine (C-C motif) receptor-like 2 (CCRL2), in the egg and the embryonic annexes is currently lacking, and their role during embryogenesis remains unknown. By immunoblot using monoclonal anti-chicken antibodies and Enzyme Linked Immunosorbent Assays (ELISA), we found that chemerin is expressed 10 times higher in albumen eggs than in blood plasma, and it is also abundant in the perivitelline membrane but undetectable in yolk. Chicken chemerin can inhibit bacterial growth. By Reverse Transcription-quantitative Polymerisation Chain Reaction (RT-qPCR), western-blot, and immunofluorescence, we show that chemerin is locally produced by the oviduct magnum that participates in albumen formation. Using cultures of magnum explants, we demonstrate that progesterone (P4) and oestradiol (E2) treatment increases chemerin secretion into cultured media and expression in magnum. Chemerin and its three receptors are present in amniotic and Chorio Allantoic Membranes (CAM). Only CMKLR1 expression decreased from embryonic day (ED) 7 to ED11 and remained low until ED18. Chemerin concentrations strongly increased in amniotic fluid at D14 when egg albumen crossed the amniotic membrane. In ovo injections of neutralising chemerin and CMKLR1 antibodies (0.01, 0.1 and 1 µg) increased embryo mortality, which occurred mainly at ED12-13, in a dose-dependent manner. Chemerin treatment increased primary CAM viability. Finally, chemerin and CMKLR1 inhibition within the CAM led to a decrease in blood vessel development and associated angiogenic gene expression. Our results show an important function of the chemerin system during embryo development in chickens, suggesting the potential use of this adipokine as a predictive marker for egg fertility or hatchability.
目前对于卵和胚胎附属物中趋化素及其受体(趋化素样受体 1(CMKLR1)、G 蛋白偶联受体 1(GPR1)和趋化因子(C-C 基序)受体样 2(CCRL2)的分布和功能知之甚少。通过使用单克隆抗鸡抗体进行免疫印迹和酶联免疫吸附试验(ELISA),我们发现,与血浆相比,卵清中趋化素的表达水平高出 10 倍,并且在卵周隙膜中也很丰富,但在蛋黄中无法检测到。鸡趋化素可以抑制细菌生长。通过反转录定量聚合酶链反应(RT-qPCR)、western-blot 和免疫荧光,我们表明趋化素是由参与卵清形成的输卵管峡部局部产生的。通过对峡部外植体的培养,我们证明孕激素(P4)和雌二醇(E2)处理增加了培养物中趋化素的分泌和峡部中的表达。趋化素及其三个受体存在于羊膜和绒毛尿囊膜(CAM)中。只有 CMKLR1 的表达从胚胎第 7 天(ED)到第 11 天下降,并一直保持低水平直到第 18 天。当卵白蛋白穿过羊膜时,D14 时羊水中的趋化素浓度强烈增加。在鸡胚中注射中和趋化素和 CMKLR1 抗体(0.01、0.1 和 1μg)会导致胚胎死亡率增加,这种情况主要发生在 ED12-13 时,且呈剂量依赖性。趋化素处理增加了初级 CAM 的活力。最后,CAM 内趋化素和 CMKLR1 的抑制作用导致血管发育减少和相关血管生成基因表达减少。我们的研究结果表明,趋化素系统在鸡胚胎发育中具有重要功能,提示该脂肪因子可作为预测卵的生育力或孵化率的指标。