Berisha B, Schams D, Rodler D, Sinowatz F, Pfaffl M W
Faculty of Agriculture and Veterinary, University of Prishtina, Prishtinë, Kosovo.
Institute of Animal Physiology and Immunology Weihenstephan, Technical University of Munich, Freising, Germany.
Reprod Domest Anim. 2017 Feb;52(1):130-139. doi: 10.1111/rda.12867. Epub 2016 Nov 11.
The aim of this study was to characterize expression patterns of hypoxia-inducible factor-1alpha (HIF1A) and vasohibin family members (VASH1 and VASH2) during different stages of ovarian function in cow. Experiment 1: Antral follicle classification occurred by follicle size and estradiol-17beta (E2) concentration in the follicular fluid into 5 groups (<0.5, 0.5-5, 5-40, 40-180 and >180 E2 ng/ml). Experiment 2: Corpora lutea (CL) were assigned to the following stages: days 1-2, 3-4, 5-7, 8-12, 13-16 and >18 (after regression) of oestrous cycle and of pregnancy (months 1-2, 3-4, 6-7, >8). Experiment 3: Cows on days 8-12 were injected with a prostaglandin F2alpha (PGF) analogue and CL were collected before and 0.5, 2, 4, 12, 24, 48 and 64 hr after PGF injection. Expression of mRNA was measured by qPCR, steroid hormone concentration by EIA and localization by immunohistochemistry. HIF1A mRNA expression in our study increases significantly in follicles during final maturation. The highest HIF1A mRNA expression was detected during the early luteal phase, followed by a significant decrease afterwards. In contrast, the mRNA of vasohibins in small follicle was high, followed by a continuous and significant downregulation in preovulatory follicles. The obtained results show a remarkable inverse expression and localization pattern of HIF1A and vasohibins during different stages of ovarian function in cow. These results lead to the assumption that the examined factors are involved in the local mechanisms regulating angiogenesis and that the interactions between proangiogenic (HIF1A) and antiangiogenic (vasohibins) factors impact all stages of bovine ovary function.
本研究的目的是表征奶牛卵巢功能不同阶段低氧诱导因子-1α(HIF1A)和血管抑制素家族成员(VASH1和VASH2)的表达模式。实验1:根据卵泡大小和卵泡液中雌二醇-17β(E2)浓度将窦卵泡分为5组(<0.5、0.5 - 5、5 - 40、40 - 180和>180 E2 ng/ml)。实验2:将黄体(CL)分为发情周期和妊娠的以下阶段:发情周期第1 - 2天、3 - 4天、5 - 7天、8 - 12天、13 - 16天以及>18天(退化后),以及妊娠1 - 2个月、3 - 4个月、6 - 7个月、>8个月。实验3:对处于第8 - 12天的奶牛注射前列腺素F2α(PGF)类似物,并在注射PGF前以及注射后0.5、2、4、12、24、48和64小时收集黄体。通过qPCR测量mRNA表达,通过酶免疫分析测量类固醇激素浓度,并通过免疫组织化学进行定位。在我们的研究中,HIF1A mRNA表达在卵泡最终成熟期间显著增加。在黄体早期检测到最高的HIF1A mRNA表达,随后显著下降。相反,血管抑制素的mRNA在小卵泡中含量很高,随后在排卵前卵泡中持续且显著下调。所得结果显示,在奶牛卵巢功能的不同阶段,HIF1A和血管抑制素呈现出显著的反向表达和定位模式。这些结果表明,所研究的因子参与调节血管生成的局部机制,并且促血管生成因子(HIF1A)和抗血管生成因子(血管抑制素)之间的相互作用影响牛卵巢功能的所有阶段。