Liang Aixin, Salzano Angela, D'Esposito Maurizio, Comin Antonella, Montillo Marta, Yang Liguo, Campanile Giuseppe, Gasparrini Bianca
Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, PR China.
Department of Veterinary Medicine and Animal Production, Federico II University, Naples, Italy.
Theriogenology. 2016 Sep 1;86(4):963-970. doi: 10.1016/j.theriogenology.2016.03.020. Epub 2016 Mar 19.
High individual variability in follicular recruitment and hence in the number of embryos produced is a major factor limiting the application of reproductive technologies in buffalo. Therefore, the identification of reliable markers to select embryo donors is critical to enroll buffaloes in embryo production programs. Better understanding of factors involved in follicular growth is also necessary to improve the response to superovulation in this species. The aim of this work was thus to determine the anti-Mullerian hormone (AMH) concentration in follicular fluid (FF) recovered from different size follicles and evaluate the mRNA expression profiles of development-related (AMHR2, CYP19A1, FSHR, and LHR) and apoptosis-related genes (TP53INP1 and CASP3) in the corresponding granulosa cells (GCs) in buffalo. Another objective was to evaluate whether the AMH concentration in FF and gene expression of GCs is associated with the antral follicular count. Ovaries were collected at the slaughterhouse, and all follicles were counted and classified as small (3-5 mm), medium (5-8 mm), and large (>8 mm). Follicular fluid was recovered for AMH determination, and the mRNA expression of AMHR2, FSHR, LHR, CYP19A1, TP53INP1, and CASP3 was analyzed in GCs. The AMH concentration in FF decreased (P < 0.01) at increasing follicular diameter. The mRNA expression of AMHR2 and FSHR was higher (P < 0.05) in small follicles, whereas that of LHR and CYP19A1 was higher (P < 0.05) in large follicles. The intrafollicular AMH concentration was positively correlated with the antral follicular count (r = 0.31; P < 0.05). Interestingly, good donors (≥12 follicles) had a higher (P < 0.05) concentration of AMH and AMHR2 levels in small follicles and higher (P < 0.05) LHR levels in large follicles than bad donors (<12 follicles). These results suggest a potential use of AMH to select buffalo donors to enroll in embryo production programs, laying the basis for further investigations.
卵泡募集存在高度个体差异，进而导致产生的胚胎数量不同，这是限制水牛生殖技术应用的主要因素。因此，识别可靠的标记物以选择胚胎供体对于将水牛纳入胚胎生产计划至关重要。更好地了解卵泡生长所涉及的因素对于改善该物种对超数排卵的反应也很有必要。因此，本研究的目的是测定从不同大小卵泡中回收的卵泡液（FF）中的抗苗勒管激素（AMH）浓度，并评估水牛相应颗粒细胞（GCs）中发育相关基因（AMHR2、CYP19A1、FSHR和LHR）和凋亡相关基因（TP53INP1和CASP3）的mRNA表达谱。另一个目的是评估FF中的AMH浓度和GCs的基因表达是否与窦状卵泡计数相关。在屠宰场收集卵巢，对所有卵泡进行计数并分类为小卵泡（3 - 5毫米）、中卵泡（5 - 8毫米）和大卵泡（>8毫米）。回收卵泡液用于测定AMH，并分析GCs中AMHR2、FSHR、LHR、CYP19A1、TP53INP1和CASP3的mRNA表达。随着卵泡直径增加，FF中的AMH浓度降低（P < 0.01）。AMHR2和FSHR的mRNA表达在小卵泡中较高（P < 0.05），而LHR和CYP19A1的mRNA表达在大卵泡中较高（P < 0.05）。卵泡内AMH浓度与窦状卵泡计数呈正相关（r = 0.31；P < 0.05）。有趣的是，优质供体（≥12个卵泡）与劣质供体（<12个卵泡）相比，小卵泡中的AMH浓度和AMHR2水平较高（P < 0.05），大卵泡中的LHR水平较高（P < 0.05）。这些结果表明AMH在选择水牛供体纳入胚胎生产计划方面具有潜在用途，为进一步研究奠定了基础。
