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热带适应性山羊卵泡液的蛋白质组学分析

Proteomic analysis of follicular fluid from tropically-adapted goats.

作者信息

Paula Junior Alexandre R, van Tilburg Mauricio F, Lobo Marina D P, Monteiro-Moreira Ana C O, Moreira Renato A, Melo Carlos H S, Souza-Fabjan Joanna M G, Araújo Airton A, Melo Luciana M, Teixeira Dárcio I A, Moura Arlindo A, Freitas Vicente J F

机构信息

School of Veterinary Medicine, State University of Ceará, Fortaleza, Brazil.

Laboratory of Human Biochemistry, State University of Ceará, Fortaleza, Brazil.

出版信息

Anim Reprod Sci. 2018 Jan;188:35-44. doi: 10.1016/j.anireprosci.2017.11.005. Epub 2017 Nov 6.

Abstract

The present study was conducted to characterize the major proteome of ovarian follicular fluid from locally-adapted, "Canindé" goats in the northeast of Brazil. Eight estrous cycling goats received a hormonal treatment consisting of medroxyprogesterone acetate, D-cloprostenol and FSH. Fluid was collected by laparoscopy from small (<3mm), medium (3-4mm) and large (>4mm) follicles and then, proteins were analyzed by 2-D SDS-PAGE and tandem mass spectrometry. Thirty-six proteins were identified in the goat follicular fluid, including albumin, immunoglobulins, ceruloplasmin, complement factor B, alpha-1B-glycoprotein precursor, serotransferrin, complement C3 and serpins, among others. Albumin and immunoglobulins were the most abundant proteins. Protein concentrations were similar in the fluid from small (45.3±3.1mg/mL), medium (44.2±3.3mg/mL) and large follicles (45.1±2.3mg/mL). The intensities of spots identified in 2-D gels as serotransferrin, zinc-alpha-2-glycoprotein-like, complement factor B and complement protein C3 differed (P<0.05) among follicle categories. The amount of serotransferrin was greater in the medium than small follicles (P<0.05). Content of zinc-alpha-2-glycoprotein-like, complement factor B and complement C3 was greater (P<0.05) in the fluid of large follicles than in medium follicles. Based on gene ontology, the major molecular functions associated with goat follicular fluid proteins were binding and catalytic activity, while the main biological processes were related to regulation, cellular processing, location and the immune system. In conclusion, the major proteome of the follicular fluid from goats subjected to hormonal stimulation was elucidated in the present study. Also, molecules associated with follicle development are potential biomarkers of oocyte competence were prevalent.

摘要

本研究旨在对巴西东北部本地适应品种“卡宁德”山羊卵泡液的主要蛋白质组进行表征。八只处于发情周期的山羊接受了由醋酸甲羟孕酮、氯前列醇D和促卵泡素组成的激素处理。通过腹腔镜从直径小于3mm的小卵泡、直径3 - 4mm的中等卵泡和直径大于4mm的大卵泡中采集卵泡液,然后通过二维SDS - PAGE和串联质谱对蛋白质进行分析。在山羊卵泡液中鉴定出36种蛋白质,包括白蛋白、免疫球蛋白、铜蓝蛋白、补体因子B、α-1B-糖蛋白前体、血清转铁蛋白、补体C3和丝氨酸蛋白酶抑制剂等。白蛋白和免疫球蛋白是含量最丰富的蛋白质。小卵泡(45.3±3.1mg/mL)、中等卵泡(44.2±3.3mg/mL)和大卵泡(45.1±2.3mg/mL)卵泡液中的蛋白质浓度相似。二维凝胶中鉴定为血清转铁蛋白、锌-α-2-糖蛋白样、补体因子B和补体蛋白C3的斑点强度在不同卵泡类别之间存在差异(P<0.05)。血清转铁蛋白在中等卵泡中的含量高于小卵泡(P<0.05)。锌-α-2-糖蛋白样、补体因子B和补体C3在大卵泡液中的含量高于中等卵泡(P<0.05)。基于基因本体论,与山羊卵泡液蛋白质相关的主要分子功能是结合和催化活性,而主要生物学过程与调节、细胞加工、定位和免疫系统有关。总之,本研究阐明了经激素刺激的山羊卵泡液的主要蛋白质组。此外,与卵泡发育相关的分子是卵母细胞能力的潜在生物标志物,且普遍存在。

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