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生长分化因子9和骨形态发生蛋白15对犬卵母细胞体外成熟的影响。

Influence of growth differentiation factor 9 and bone morphogenetic protein 15 on in vitro maturation of canine oocytes.

作者信息

Garcia Pablo, Aspee Karla, Ramirez Georges, Dettleff Phillip, Palomino Jaime, Peralta Oscar A, Parraguez Víctor H, De Los Reyes Monica

机构信息

Laboratory of Animal Reproduction, Department of Animal Production, Faculty of Veterinary Sciences, University of Chile, Santiago, Chile.

Laboratory of Animal Physiology, Department of Biological Sciences, Faculty of Veterinary Sciences, University of Chile, Santiago, Chile.

出版信息

Reprod Domest Anim. 2019 Feb;54(2):373-380. doi: 10.1111/rda.13371. Epub 2018 Dec 3.

Abstract

Growth differentiation factor 9 (GDF-9) and bone morphogenetic protein 15 (BMP-15) have pivotal roles in oocyte development in many species, therefore the aim was to investigate these factors during in vitro maturation (IVM) of canine oocytes. Canine cumulus oocytes complexes (COCs) were cultured in six groups for 72 hr in a supplemented TCM199-Hepes medium as (a) Control group; (b) GDF-9 antibody (Ab); (c) BMP-15 Ab; (d) recombinant human (rh) GDF-9; (e) rh BMP-15 or (f) rh BMP-15 and GDF-9. Data were evaluated by ANOVA. The Abs against GDF-9 or BMP-15 had a negative impact on meiotic development. Higher (p < 0.05) number of oocytes was arrested at GVBD stage when they were incubated with either GDF-9 Ab (64.4 ± 2.1%) or BMP-15 Ab (67.2%± 4.9%) in comparison to those in control group (32.4 ± 7.8%). In contrast, more (p < 0.05) oocytes in control group reached MI (37.4 ± 1.3%) and MII stages (10.2 ± 2.1%) comparing to those groups with GDF-9 Ab (23.1 ± 4.7% MI; 0.0% MII) or BMP-15 Ab (16.4 ± 2.4%MI; 5.9% ± 2.1 MII). Higher rates (p < 0.05) of oocytes in control group stayed still arrested at GV (19.9 ± 8.6%) in comparison to those cultured with either rhGDF-9 (3.7 ± 0.4%) or rhBMP-15 (10.9 ± 0.7%). However, there were no differences in MII rates between oocytes cultured with GDF-9 (14.7 ± 3.1) and BMP-15 (7.8 ± 2.5) separately. But, more oocytes (p < 0.05) reached the MII stage (20.5 ± 3.8%) compared to those exposed to each protein separately and to the control group. These results suggest that these proteins likely contribute to the meiotic development in dogs.

摘要

生长分化因子9(GDF-9)和骨形态发生蛋白15(BMP-15)在许多物种的卵母细胞发育中起关键作用,因此本研究旨在探讨犬卵母细胞体外成熟(IVM)过程中的这些因子。犬卵丘卵母细胞复合体(COCs)在添加了TCM199-Hepes的培养基中分为六组培养72小时,分别为:(a)对照组;(b)GDF-9抗体(Ab)组;(c)BMP-15 Ab组;(d)重组人(rh)GDF-9组;(e)rh BMP-15组;(f)rh BMP-15和GDF-9组。数据采用方差分析进行评估。抗GDF-9或BMP-15的抗体对减数分裂发育有负面影响。与对照组(32.4±7.8%)相比,当卵母细胞与GDF-9 Ab(64.4±2.1%)或BMP-15 Ab(67.2%±4.9%)孵育时,更多(p<0.05)的卵母细胞停滞在GVBD期。相反,与GDF-9 Ab组(23.1±4.7%MI;0.0%MII)或BMP-15 Ab组(16.4±2.4%MI;5.9%±2.1%MII)相比,对照组更多(p<0.05)的卵母细胞达到MI期(37.4±1.3%)和MII期(10.2±2.1%)。与用rhGDF-9(3.7±0.4%)或rhBMP-15(10.9±0.7%)培养的卵母细胞相比,对照组中停滞在GV期的卵母细胞比例更高(p<0.05)(19.9±8.6%)。然而,单独用GDF-9(14.7±3.1)和BMP-15(7.8±2.5)培养的卵母细胞在MII期的比例没有差异。但是,与单独暴露于每种蛋白质的组和对照组相比,更多(p<0.05)的卵母细胞达到了MII期(20.5±3.8%)。这些结果表明,这些蛋白质可能有助于犬类的减数分裂发育。

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