Ilha G F, Rovani M T, Gasperin B G, Ferreira R, de Macedo M P, Neto O A, Duggavathi R, Bordignon V, Gonçalves P B D
Laboratory of Biotechnology and Animal Reproduction - BioRep, Federal University of Santa Maria, Santa Maria, Brazil.
Laboratory of Animal Reproduction - ReproPEL, Federal University of Pelotas, Pelotas, Brazil.
Reprod Domest Anim. 2016 Apr;51(2):188-94. doi: 10.1111/rda.12662. Epub 2016 Jan 27.
The anti-Müllerian hormone (AMH) is an important marker of ovarian reserve and for predicting the response to superovulatory treatments in several species. The objective of this study was to investigate whether AMH and its receptor (AMHR2) are regulated in bovine granulosa cells during follicular development. In the first experiment, granulosa cells were retrieved from the two largest follicles on days 2 (before), 3 (at the expected time) or 4 (after deviation) of follicular wave. In the second experiment, four doses of FSH (30, 30, 20 and 20 mg) or saline were administered twice a day starting on Day 2 of the first follicular wave of the cycle. Granulosa cells and follicular fluid were collected from the two largest follicles 12 h after the last injection of FSH or saline. AMH mRNA abundance was similar in granulosa cells of the two largest follicles (F1 and F2) before deviation (Day 2), but greater in dominant (DF) than subordinate follicles (SF) at the expected time (Day 3) and after (Day 4) deviation (p < 0.05). In experiment 1, AMH mRNA levels declined in both DF and SF near the expected time and after deviation when compared to before deviation. There was no difference in AMHR2 mRNA levels before and during follicular deviation (p > 0.05), but they tended to be greater in DFs than SFs (p < 0.1) after deviation. Experiment 2 showed that AMH and AMHR2 mRNA in granulosa cells and AMH protein abundance in follicular fluid were similar (p > 0.05) between both co-dominant follicles collected from the FSH-treated cows. These findings indicate the followings: AMH mRNA levels decrease in both DFs and SFs during follicular deviation; granulosa cells from heathy follicles express more AMH mRNA compared to subordinate follicles undergoing atresia and FSH stimulates AMH and AMHR2 mRNA expression in granulosa cells of co-dominant follicles.
抗苗勒管激素(AMH)是卵巢储备的重要标志物,也是预测多种物种对超排卵治疗反应的指标。本研究的目的是调查在卵泡发育过程中,牛颗粒细胞中的AMH及其受体(AMHR2)是否受到调控。在第一个实验中,在卵泡波的第2天(之前)、第3天(预期时间)或第4天(偏离后)从两个最大的卵泡中获取颗粒细胞。在第二个实验中,从周期的第一个卵泡波的第2天开始,每天两次给予四剂促卵泡素(FSH,分别为30、30、20和20mg)或生理盐水。在最后一次注射FSH或生理盐水12小时后,从两个最大的卵泡中收集颗粒细胞和卵泡液。在偏离前(第2天),两个最大卵泡(F1和F2)的颗粒细胞中AMH mRNA丰度相似,但在预期时间(第3天)和偏离后(第4天),优势卵泡(DF)中的AMH mRNA丰度高于从属卵泡(SF)(p<0.05)。在实验1中与偏离前相比,在预期时间附近和偏离后,DF和SF中的AMH mRNA水平均下降。在卵泡偏离之前和期间,AMHR2 mRNA水平没有差异(p>0.05),但在偏离后,DF中的AMHR2 mRNA水平往往高于SF(p<0.1)。实验2表明,从接受FSH治疗的奶牛收集的两个共同优势卵泡之间,颗粒细胞中的AMH和AMHR2 mRNA以及卵泡液中的AMH蛋白丰度相似(p>0.05)。这些发现表明:在卵泡偏离过程中,DF和SF中的AMH mRNA水平均下降;与正在闭锁的从属卵泡相比,健康卵泡的颗粒细胞表达更多的AMH mRNA,并且FSH刺激共同优势卵泡的颗粒细胞中AMH和AMHR2 mRNA的表达。