Bastos H B A, Martinez M N, Camozzato G C, Estradé M J, Barros E, Vital C E, Vidigal P M P, Meikle A, Jobim M I M, Gregory R M, Mattos R C
REPROLAB - Faculdade de Veterinária, UFRGS, Porto Alegre, RS, Brazil.
Facultad de Veterinária, UDELAR, Montevideo, Uruguay.
Theriogenology. 2019 Feb;125:224-235. doi: 10.1016/j.theriogenology.2018.11.002. Epub 2018 Nov 17.
There is a complex cascade involving proteins during early embryo development and maternal recognition, which is very important for maintenance of a conceptus. The aim of this study was to compare proteomic profile of uterine fluid after ovulation in pregnant and cyclic mares. In the first cycle, samples of uterine fluid of 30 cyclic mares were collected on days 7 (n = 10), 10 (n = 10) and 13 (n = 10) post ovulation and constituted the Cyclic group. In the second cycle, the same mares were bred to a fertile stallion. At days 7, 10 and 13 uterine fluid samples were collected. Immediately after sample collection, the mare's uteri were flushed, and those with an embryo recovered were assigned to the Pregnant group. Of the 30 mares flushed embryos were recovered from 6 mares on day 7, 6 on day 10 and 6 on day 13. Samples from the mares without embryo recovery were excluded from both groups. The uterine fluid samples were processed by two-dimensional electrophoresis technique followed by matrix assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) mass spectrometry for the identification of relevant protein spots. From a total of 677 detected spots 19 were identified, 13 more abundant in Pregnant group and 6 in Cyclic group. In summary, pregnant and cyclic mares showed proteins with different abundance. Identified proteins were related to the transport of lipids through the embryo capsule, uterine motility, ATP generation, maternal immunological tolerance, cell proliferation, differentiation, metabolism and angiogenesis. Changes in the proteomic profile of uterine fluid during early embryo development in mares were related with the conceptus presence, suggesting that these alterations may be important for conceptus development and maternal recognition of pregnancy.
在早期胚胎发育和母体识别过程中存在一个涉及蛋白质的复杂级联反应，这对于维持孕体非常重要。本研究的目的是比较怀孕母马和发情周期母马排卵后子宫液的蛋白质组学特征。在第一个周期，于排卵后第7天（n = 10）、第10天（n = 10）和第13天（n = 10）采集30匹发情周期母马的子宫液样本，构成发情周期组。在第二个周期，让同一批母马与一匹能育种马交配。在第7天、第10天和第13天采集子宫液样本。样本采集后立即冲洗母马子宫，回收有胚胎的母马并将其归入怀孕组。在30匹母马中，第7天从6匹母马、第10天从6匹母马以及第13天从6匹母马中回收了胚胎。未回收胚胎的母马样本从两组中排除。子宫液样本采用二维电泳技术处理，随后进行基质辅助激光解吸/电离飞行时间/飞行时间（MALDI-TOF/TOF）质谱分析以鉴定相关蛋白质斑点。在总共检测到的677个斑点中，鉴定出19个，其中13个在怀孕组中丰度更高，6个在发情周期组中丰度更高。总之，怀孕母马和发情周期母马显示出丰度不同的蛋白质。鉴定出的蛋白质与脂质通过胚胎囊的转运、子宫运动、ATP生成、母体免疫耐受、细胞增殖、分化、代谢和血管生成有关。母马早期胚胎发育过程中子宫液蛋白质组学特征的变化与孕体的存在有关，表明这些改变可能对孕体发育和母体对妊娠的识别很重要。
