Pinto Pedro Henrique Nicolau, Balaro Mario Felipe Alvarez, Souza-Fabjan Joanna Maria Gonçalves, Ribeiro Lilian Dos Santos, Bragança Gláucia Mota, Leite Ceci Ribeiro, Arashiro Eduardo Kenji Nunes, de Moraes Silva Kleibe, Da Fonseca Jeferson Ferreira, Brandão Felipe Zandonadi
Faculdade de Veterinária, Universidade Federal Fluminense, Rua Vital Brazil Filho, 64, CEP 24230-340, Niterói, RJ, Brazil.
Faculdade de Veterinária, Universidade Federal Fluminense, Rua Vital Brazil Filho, 64, CEP 24230-340, Niterói, RJ, Brazil.
Theriogenology. 2018 Jun;113:146-152. doi: 10.1016/j.theriogenology.2018.02.018. Epub 2018 Feb 23.
This study aims to compare four different methods for selecting high responding sheep donors for in vivo embryo production. These methods include a pre-selection eCG test (eCG), antral follicle count (AFC), plasma anti-Müllerian hormone measurement (AMH) and genotyping for the presence of the FecG mutation (a polymorphism in the GDF9 gene associated with increased ovulation rate). Santa Ines ewe lambs (n = 25) underwent superovulation (SOV) with 800 IU equine chorionic gonadotropin (eCG) and the corpus luteum (CL) count was recorded by laparoscopy after eight days. At the D0, blood samples for AMH and genotyping analysis were collected. Twenty-one days after the end of the eCG test, the same animals underwent SOV with 200 mg of FSH, administered in six decreasing doses, and then naturally mated. Immediately before the beginning of the FSH protocol (D0), and at the moment of the first FSH dose (D9), the AFC was assessed. Plasma AMH was again determined at the D9. After each screening process, animals were classified as having a high (HR), or low (LR), potential of response (using specific thresholds for each method). Then, the ewes' response to SOV and embryo yield for each screening method, classified as HR or LR, were compared. Animals classified as HR by AFC (HR) and by AMH concentration (HR) at the D9, produced more viable embryos than those classified as LR and LR (HR 6.2 ± 3.2 vs LR 2.8 ± 3.0 and HR 6.6 ± 3.6 vs LR 3.0 ± 2.9). Pre-selection tests with eCG and different FecG genotypes, either heterozygous (+/E) or wild type (+/+), were unable to discriminate HR or LR animals. A tendency (P = 0.06) to have lower plasma AMH was observed in heterozygous FecG (+/E) ewes. In conclusion, both AFC and plasma AMH can be used to select donor ewes with a higher potential of response for in vivo embryo production.
本研究旨在比较四种不同的方法，用于选择体内胚胎生产中高反应性的绵羊供体。这些方法包括预筛选马绒毛膜促性腺激素（eCG）试验（eCG）、窦卵泡计数（AFC）、血浆抗苗勒管激素测定（AMH）以及检测FecG突变（生长分化因子9基因中与排卵率增加相关的一种多态性）的基因分型。25只圣伊内斯母羊羔羊用800国际单位马绒毛膜促性腺激素（eCG）进行超排，8天后通过腹腔镜记录黄体（CL）数量。在第0天，采集血样用于AMH和基因分型分析。eCG试验结束21天后，相同的动物用200毫克促卵泡素（FSH）进行超排，以六种递减剂量给药，然后自然交配。在FSH方案开始前即刻（第0天）以及首次注射FSH时（第9天）评估AFC。在第9天再次测定血浆AMH。每次筛选过程后，根据每种方法的特定阈值，将动物分为高反应性（HR）或低反应性（LR）。然后，比较每种筛选方法分类为HR或LR的母羊对超排的反应和胚胎产量。在第9天通过AFC分类为HR和通过AMH浓度分类为HR的动物，产生的活胚比分类为LR和LR的动物更多（HR 6.2±3.2对LR 2.8±3.0以及HR 6.6±3.6对LR 3.0±2.9）。用eCG和不同FecG基因型（杂合子（+/E）或野生型（+/+））进行的预筛选试验无法区分HR或LR动物。在杂合子FecG（+/E）母羊中观察到血浆AMH有降低的趋势（P = 0.06）。总之，AFC和血浆AMH均可用于选择体内胚胎生产中具有较高反应潜力的供体母羊。
