Camacho Cesar Augusto, Santos Gabriel de Oliveira, Caballeros Jorge Emilio, Cazales Nicolas, Ramirez Camilo José, Vidigal Pedro Marcus Pereira, Ramos Humberto Josué de Oliveira, Barros Edvaldo, Mattos Rodrigo Costa
Laboratório de Reprodução Animal, Faculdade de Veterinária, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brasil.
Facultad de Veterinária, Universidad de la República - UDELAR, Montevideo, Uruguay.
Anim Reprod. 2020 Nov 25;17(4):e20200552. doi: 10.1590/1984-3143-AR2020-0552.
This experiment aimed to compare at day seven after ovulation, the protein profile of uterine fluid in cyclic mares with mares infused two days before with Day 13 conceptus fragments. Experimental animals were ten healthy cyclic mares, examined daily to detect ovulation (Day 0) as soon as estrus was confirmed. On day seven, after ovulation, uterine fluid was collected, constituting the Cyclic group (n = 10). The same mares were examined in the second cycle until ovulation was detected. On day five, after ovulation, fragments from a previously collected concepti were infused into each mare's uterus. Two days after infusion, uterine fluid was collected, constituting the Fragment group (n = 10). Two-dimensional electrophoresis technique processed uterine fluid samples. A total of 373 spots were detected. MALDI-TOF/TOF and NanoUHPLC-QTOF mass spectrometry identified twenty spots with differences in abundance between the Cyclic and Fragment group. Thirteen proteins were identified, with different abundance between groups. Identified proteins may be related to embryo-maternal communication, which involves adhesion, nutrition, endothelial cell proliferation, transport, and immunological tolerance. In conclusion, conceptus fragments signalized changes in the protein profile of uterine fluid seven days after ovulation in comparison to the observed at Day 7 in the same cyclic mares.
本实验旨在比较排卵后第7天,发情周期母马与在排卵前两天注入第13天孕体碎片的母马子宫液中的蛋白质谱。实验动物为10匹健康的发情周期母马,每天进行检查,一旦确认发情就检测排卵(第0天)。在排卵后的第7天,收集子宫液,构成发情周期组(n = 10)。在第二个发情周期对同一批母马进行检查,直至检测到排卵。在排卵后的第5天,将先前收集的孕体碎片注入每匹母马的子宫。注入后两天,收集子宫液,构成碎片组(n = 10)。采用二维电泳技术处理子宫液样本。共检测到373个斑点。基质辅助激光解吸电离飞行时间串联质谱(MALDI-TOF/TOF)和纳升级超高效液相色谱-四极杆飞行时间质谱(NanoUHPLC-QTOF)鉴定出20个在发情周期组和碎片组中丰度存在差异的斑点。鉴定出13种蛋白质,其在两组间丰度不同。鉴定出的蛋白质可能与胚胎-母体通讯有关,这涉及黏附、营养、内皮细胞增殖、运输和免疫耐受。总之,与同一发情周期母马在第7天观察到的情况相比,孕体碎片显示出排卵后第7天子宫液蛋白质谱的变化。