Jimenez C R, de Azevedo J L, Silveira R G, Penitente-Filho J, Carrascal-Triana E L, Zolini A M, Araújo V R, Torres Caa
Departament of Animal Science, Laboratory of Animal Physiology and Reproduction, Universidade Federal de Viçosa, Viçosa, MG, Brazil.
Laboratory of Manipulation of Oocytes Enclosed in Preantral Follicles, Universidade Estadual do Ceara, Fortaleza, CE, Brazil.
Reprod Domest Anim. 2016 Aug;51(4):575-84. doi: 10.1111/rda.12722. Epub 2016 Jun 25.
The objective of this study was to evaluate different concentrations of growth hormone (GH) on the development of bovine preantral follicles cultured included in the ovarian tissue (in situ) on the rates of morphologically normal, viable, primordial and developing follicles, as well as the oocyte and follicle diameter and ultrastructural analysis. Ovarian fragments collected from cows with no cross-breeds defined were cultured in situ for 1 and 7 days in minimal essential medium (α-MEM+) supplemented with different concentrations of recombinant human GH (0, 10, 25, 50 ng/ml). The ovarian fragments non-cultured (control) and cultured were processed for classic histology, mechanical isolation and electron transmission microscopy (MET). The parameters underwent anova (Tukey's and Dunnett's tests) and chi-square test (χ(2) ). After 7 days of culture, the treatment with 50 ng/ml GH showed no differences with fresh control (p > 0.05) and had greater effectiveness than in the 0, 10 and 25 ng/ml GH concentrations of the morphologically normal follicles. Regarding the primordial follicles, a reduction was observed in the 50 ng/ml GH concentration concomitant with the significant increase in developing follicles, differing from both the fresh control and the other GH concentrations tested. In addition, 50 ng/ml GH showed a larger follicle and oocyte diameter when compared to the other treatments cultured. Similar structures were ultrastructurally observed in the control group, 50 ng/ml GH. Follicles cultured in 10 ng/ml GH showed nuclear invagination, vacuoles and lesioned basal membrane. Hence, it is concluded that 50 ng/ml GH is the most effective concentration for the development of preantral follicles cultured in situ.
本研究的目的是评估不同浓度的生长激素（GH）对培养于卵巢组织（原位）中的牛窦前卵泡发育的影响，包括形态正常、存活、原始和发育中卵泡的比率，以及卵母细胞和卵泡直径及超微结构分析。从无杂交品种定义的奶牛采集的卵巢片段在添加不同浓度重组人生长激素（0、10、25、50 ng/ml）的最低必需培养基（α-MEM+）中进行原位培养1天和7天。未培养（对照）和培养后的卵巢片段进行经典组织学、机械分离和电子透射显微镜（MET）处理。对参数进行方差分析（Tukey检验和Dunnett检验）和卡方检验（χ(2)）。培养7天后，50 ng/ml GH处理组与新鲜对照组无差异（p>0.05），且在形态正常卵泡方面比0、10和25 ng/ml GH浓度更有效。关于原始卵泡，在50 ng/ml GH浓度下观察到减少，同时发育中卵泡显著增加，这与新鲜对照组和其他测试的GH浓度均不同。此外，与其他培养处理相比，50 ng/ml GH显示卵泡和卵母细胞直径更大。在对照组、50 ng/ml GH组超微结构观察到相似结构。在10 ng/ml GH中培养的卵泡显示核内陷、空泡和基底膜损伤。因此，得出结论，50 ng/ml GH是原位培养窦前卵泡发育的最有效浓度。
