Schilffarth Stefanie, Antoni Bernadette, Schams Dieter, Meyer Heinrich H D, Berisha Bajram
Physiology Weihenstephan, Technical University Munich, Freising, Germany.
Int J Biol Sci. 2009 May 13;5(4):344-50. doi: 10.7150/ijbs.5.344.
Recent studies implicate that apelin and its receptor APJ may have important role for the modulation of angiogenesis. The aim of this study was to further characterise the regulation of apelin/APJ system in bovine ovary. Experiment 1: corpora lutea (CL) were assigned to the following stages: days 1-2, 3-4, 5-7, 8-12, 13-16, >18 (after regression) of oestrous cycle and of gravidity (month <3, 3-5, 6-7 and >8). Experiment 2: Follicles during maturation were divided into granulosa cells (GC) and theca interna (TI) and were examined separately. Classification of follicles occurred by follicle size and oestradiol-17beta (E2) concentration in the follicular fluid (FF) (<0.5 ng/ml, 0.5-5 ng/ml; 5-40 ng/ml; 40-180 ng/ml; >180 ng/ml). Real-time RT-PCR (qPCR) was applied to investigate mRNA expression of examined factors. In general, the expression level of apelin during the oestrous cycle was significantly higher compared to the one during pregnancy. Apelin mRNA levels were always high during the cycle with a tendency of decrease after CL regression. The APJ mRNA in the CL was significantly up regulated on days 5-7 and 8-12 followed by a decrease on days 13-16, and further on days >18. The expression of APJ does not show any significant regulation in the CL throughout pregnancy. The expression of apelin and APJ was not statistically regulated in GC, but was significantly up regulated in follicles with an E2 concentration of more than 5 ng/ml and showed an increase according to growth and maturation of follicles. In conclusion, our data suggest that apelin/APJ system is involved in the mechanism regulating angiogenesis during follicle maturation as well as during CL formation and function in the bovine ovary.
近期研究表明,阿片肽及其受体APJ可能在血管生成调节中发挥重要作用。本研究旨在进一步阐明牛卵巢中阿片肽/APJ系统的调控机制。实验1:将黄体(CL)分为发情周期和妊娠期的以下阶段:发情周期的第1 - 2天、3 - 4天、5 - 7天、8 - 12天、13 - 16天、>18天(退化后),以及妊娠期的<3个月、3 - 5个月、6 - 7个月和>8个月。实验2:将成熟卵泡分为颗粒细胞(GC)和内膜细胞(TI)并分别进行检测。根据卵泡大小和卵泡液(FF)中雌二醇-17β(E2)浓度(<0.5 ng/ml、0.5 - 5 ng/ml、5 - 40 ng/ml、40 - 180 ng/ml、>180 ng/ml)对卵泡进行分类。应用实时逆转录聚合酶链反应(qPCR)检测相关因子的mRNA表达。总体而言,发情周期中阿片肽的表达水平显著高于妊娠期。发情周期中阿片肽mRNA水平始终较高,黄体退化后有下降趋势。黄体中APJ mRNA在第5 - 7天和8 - 12天显著上调,随后在第13 - 16天下降,并在>18天时进一步下降。整个妊娠期黄体中APJ的表达未显示出任何显著调控。阿片肽和APJ在颗粒细胞中的表达无统计学调控,但在E2浓度超过5 ng/ml的卵泡中显著上调,并随卵泡生长和成熟而增加。总之,我们的数据表明,阿片肽/APJ系统参与了牛卵巢卵泡成熟以及黄体形成和功能过程中血管生成的调控机制。