Lee Eunyoung, Kim Sang Hwan, Kim Byung-Gak, Yoon Jong Taek
Dept. of Animal Life Science, Hankyong National University, Ansung 456-749, Korea.
Dev Reprod. 2015 Dec;19(4):227-34. doi: 10.12717/DR.2015.19.4.227.
The objective of this study was to investigate the expression of bovine luteum expressed sequence tags (ESTs), vascular endothelial growth factor (VEGF), and tumor necrosis factor receptor 1 (TNFR1) and the presence of functional ESTs in the bovine corpus luteum (CL) during different stages of the estrus cycle. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed a difference in the expression of ESTs during the CL stage. Concentration of ESTs in the CL tissue increased significantly from the mid-luteal stage and decreased thereafter. RT-PCR analysis showed higher levels of the EST genes in the CL of the mid-luteal stage than in other stages, and the same level of expression of VEGF. Immunohistochemistry analysis of the tissue from CL formation to regression showed low cytosol and aggregation of the nucleus. And activity caspase 3 (apoptosis detector) was most strongly detected in the CL1 stage of bovine. During the estrous cycle, the cytosol was magnified and differentiation of the nucleus was clearly manifested. The ESTs affected the CL, and the relationship between VEGF and TNFR1 played a pivotal role for CL development and activation, dependent on the stage of CL. These results suggest local production of ESTs, the presence of functional ESTs in the bovine CL, and that ESTs play a role in regulating the function of cell death in bovine CL.
本研究的目的是调查发情周期不同阶段牛黄体表达序列标签(EST)、血管内皮生长因子(VEGF)和肿瘤坏死因子受体1(TNFR1)的表达情况,以及牛黄体(CL)中功能性EST的存在情况。逆转录-聚合酶链反应(RT-PCR)分析显示,在黄体阶段EST的表达存在差异。黄体组织中EST的浓度从中黄体期开始显著增加,随后下降。RT-PCR分析显示,中黄体期黄体中EST基因的水平高于其他阶段,VEGF的表达水平相同。对从黄体形成到退化的组织进行免疫组织化学分析,结果显示细胞质较少且细胞核聚集。并且在牛的CL1阶段检测到活性半胱天冬酶3(凋亡检测指标)最为强烈。在发情周期中,细胞质放大,细胞核分化明显。EST影响黄体,VEGF和TNFR1之间的关系对黄体的发育和激活起着关键作用,这取决于黄体的阶段。这些结果表明牛黄体中EST的局部产生、功能性EST的存在,并且EST在调节牛黄体细胞死亡功能中发挥作用。