de Sousa Regivaldo Vieira, da Silva Cardoso Célia Regina, Butzke Guilberth, Dode Margot Alves Nunes, Rumpf Rodolfo, Franco Maurício Machaim
School of Agriculture and Veterinary Medicine, University of Brasilia, Brasília, DF, Brazil; Embrapa Genetic Resources and Biotechnology, Laboratory of Animal Reproduction, Brasília, DF, Brazil.
Guilberth Serviços Veterinários S/A, São Paulo, SP, Brazil.
Theriogenology. 2017 Mar 1;90:25-31. doi: 10.1016/j.theriogenology.2016.11.003. Epub 2016 Nov 11.
Assisted reproductive techniques have significantly contributed to animal breeding programs. Similarly, genomics has provided important information and tools to improve the accuracy of selection. However, the greatest benefits of those tools can only be expected when they are combined, allowing animals to be selected accurately early in life. Therefore, obtaining DNA samples from embryos without compromising their viability is essential for the consolidation of preimplantation genomic selection. We aimed to evaluate the effect on the gestation rate of conducting a biopsy of in vivo (VV) and in vitro-produced (IVP) bovine embryos. The VV and IVP embryos were distributed into two groups: VV-B (biopsied embryos; n = 380) and VV-C (intact embryos-controls; n = 229) and IVP-B (biopsied embryos; n = 91) and IVP-C (intact embryos-controls; n = 227), respectively. After biopsy, embryos from both groups VV-B and IVP-B were cultured for an additional 3 hours before being transferred to synchronized recipients. To evaluate the quality of the DNA obtained in the biopsies, this was used to determine the sex of embryos by polymerase chain reaction. No effect (P > 0.05) of the biopsy was observed for any of the treatments, the pregnancy rate at D 60 post-transfer being similar for VV-B: 206/380 (54.21%) and VV-C: 128/229 (55.89%) and for IVP-B: 24/91 (26.37%) and IVP-C: 45/227 (19.82%). Also, no effect (P > 0.05) of the embryo's stage of development was detected on percentage of pregnant recipients when in vitro embryos were transferred. From the biopsies analyzed, about 90% had the sex determined, confirming that DNA was there and it was efficiently amplified. The results indicated that biopsy does not affect the viability of IVV and IVP bovine embryos and can be used in commercial programs to associate assisted reproductive technologies with genomic selection.
辅助生殖技术对动物育种计划做出了重大贡献。同样,基因组学为提高选择准确性提供了重要信息和工具。然而,只有将这些工具结合起来,才能在动物生命早期进行准确选择,从而获得最大益处。因此,在不影响胚胎活力的情况下获取胚胎的DNA样本对于巩固植入前基因组选择至关重要。我们旨在评估对体内(VV)和体外生产(IVP)的牛胚胎进行活检对妊娠率的影响。VV和IVP胚胎分别分为两组:VV-B(活检胚胎;n = 380)和VV-C(完整胚胎-对照组;n = 229)以及IVP-B(活检胚胎;n = 91)和IVP-C(完整胚胎-对照组;n = 227)。活检后,VV-B和IVP-B两组的胚胎在转移到同期受体之前再培养3小时。为了评估活检获得的DNA质量,用其通过聚合酶链反应确定胚胎性别。对于任何一种处理,均未观察到活检的影响(P>0.05),移植后60天的妊娠率在VV-B组为206/380(54.21%)和VV-C组为128/229(55.89%)之间相似,在IVP-B组为24/91(26.37%)和IVP-C组为45/227(19.82%)之间相似。此外,当移植体外胚胎时,未检测到胚胎发育阶段对妊娠受体百分比的影响(P>0.05)。在分析的活检样本中,约90%的样本确定了性别,证实了DNA的存在且能有效扩增。结果表明,活检不会影响体内和体外生产的牛胚胎的活力,可用于商业计划,将辅助生殖技术与基因组选择相结合。