摘要
为研究干扰素刺激基因IFITM1和RSAD2在奶牛早期妊娠阶段外周血中的表达规律,于人工授精后0、14、18、21和28d采集奶牛外周血,利用荧光定量PCR检测IFITM1和RSAD2的相对表达量,采用化学发光法同步检测血清中孕酮浓度;并深入分析自然发情和同期发情2种状态下青年母牛外周血中RSAD2基因的相对表达及其在18d和21d外周血白细胞亚类(淋巴细胞、NK细胞、单核细胞)中的相对表达。结果表明,妊娠牛与空怀牛血清孕酮在21d和28d呈显著差异。人工授精后14d和18dIFITM1基因在妊娠牛与空怀牛中都显著下调,但在空怀牛与妊娠牛间差异不显著。妊娠牛外周血中RSAD2于21d显著上调;18d时,RSAD2在妊娠青年牛外周血中的表达量显著高于空怀牛。进一步分析结果表明:自然发情条件下,妊娠牛外周血中RSAD2表达量显著高于空怀牛,同期发情条件下该基因在趋势上与之类似但统计学上不显著。人工授精后18d和21d,RSAD2在妊娠牛与空怀牛外周血淋巴细胞、NK细胞和单核细胞中的表达均无显著差异。研究提示:IFITM1不适合用于奶牛早期妊娠诊断;利用RSAD2的相对表达量可望对青年牛在18d做出早期诊断,具体判别标准有待于进一步扩群分析。
The present experiment aimed to estimate mRNA transcriptions of IFITM1 and RSAD2 in the peripheral blood of Chinese Holstein dairy cows during early pregnancy. Peripheral blood at five different time points （0 d, 14 d, 18 d, 21 d, and 28 d） after artificial insemination （AI） were collected. Re- al-time PCR was employed to detect the relative gene expressions of IFITM1 and RSAD2, while plasma P4 was detected by Chemiluminescence Immunoassay （CLIA） at the same time. Transcriptions of RSAD2 were evaluated in the whole blood of heifers with spontaneous cycles of estrus and/or synchroni- zation of estrus. In addition, typical peripheral blood leukocyte subsets （lymphocyte, natural killer cell, and monocyte） were isolated at 0 d,18 d,and 21 d after AI and gene transcriptions of RSAD2 were de- tected. The results showed that pregnant cows had significant higher （P〈0. 05） plasma P4 levels than non-pregnant cows at 21 d and 28 d. At 14 d and 18 d, transcriptions of bovine IFITM1 in peripheral blood were significantly declined compared with cows at 0 d （P〈0. 05）, but there was no significant difference between pregnant cows and non-pregnant cows. Although transcriptions of RSAD2 in preg- nant cows were elevated significantly at 21 d （P〈0.05）, expressions of RSAD2 in pregnant heifers were significantly higher than those in non-pregnant cows as early as 18 d. Further analysis discovered that transcriptions of RSAD2 in pregnant cows with spontaneous cycles of estrus were significant higher than those in non-pregnant cows at 18 d. There was a same trend in cows with synchronization of estrus, but the difference was not statistically significant. At 18 d and 21 d, there was no significant difference of RSAD2 expression in the blood leukocyte subsets between the pregnant cows and non-pregnant cows. These results indicated that transcriptions of bovine IFITM1 was not reliable for early pregnancy diag- nosis,while RSAD2 could be applied in bovine pregnancy diagnosis at 18 d in heifers. Further stu
出处
《华中农业大学学报》
CAS
CSCD
北大核心
2015年第3期89-96,共8页
Journal of Huazhong Agricultural University
基金
国家自然科学基金项目(31201766)
武汉市农科院人才发展专项(CX201240)
武汉市农科院青年英才计划项目(YC201201)
