首次分离和鉴定黄羊输卵管液细胞外囊泡。

IF 1.6 4区 农林科学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE
Animal Reproduction Pub Date : 2024-10-21 eCollection Date: 2024-01-01 DOI:10.1590/1984-3143-AR2024-0039
Roberto Mendes, Agostinho Soares de Alcântara, Gildas Mbemya Tetaping, Marco Aurélio Schiavo Novaes, Vanessa Barbosa Pinheiro Gonçalves, João Xavier da Silva, José Jonathas Albuquerque de Almeida, Maria Izabel Florindo Guedes, Luzia Kalyne Almeida Moreira Leal, Joanna Maria Gonçalves Souza-Fabjan, Roberto Nicolete, Deborah de Melo Magalhães Padilha, José Ricardo de Figueiredo, Ana Paula Ribeiro Rodrigues
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引用次数: 0

摘要

输卵管液细胞外囊泡(oEV)对于围受精过程至关重要。在哺乳动物的输卵管液(OF)中已经发现了EV的存在,但黄羊除外。因此,本研究旨在分离和鉴定黄裘鼠的 oEV(coEV)。在实验 1 中,首先从每只动物的输卵管液或三只动物的输卵管液中分离 coEV。在实验 2 中,在发情周期的卵泡期或黄体期分离 coEV。利用动态光散射(DLS)分析法、透射电子显微镜(TEM)和点印迹法(DB)对 coEV 的大小分布、多分散指数(PDI)和 zeta 电位进行了表征。我们的结果表明,无论采用哪种分离方法(单个或集合),coEV 的理化特性都相似(P > 0.05)。然而,黄体期采集的 coEV 比卵泡期采集的 coEV 要大(P < 0.05)。TEM 显示了外泌体特有的球形和杯形颗粒。DB 揭示了参与 coEV 生物生成的外泌体蛋白的存在。总之,可以从单个雌性山羊体内分离并鉴定 coEV,发情周期阶段会影响囊泡的平均大小和 PDI。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
First isolation and characterization of caprine oviduct fluid extracellular vesicles.

Oviduct fluid extracellular vesicles (oEV) are essential for periconceptional events. The presence of EV has already been identified in the oviduct fluid (OF) from mammalian species, except in caprine. Therefore, this study aimed to isolate and characterize the caprine oEV (coEV). Initially, in Experiment 1, coEV were isolated from the OF of either each animal individually or from a pool of three animals. In experiment 2, coEV were isolated during the follicular or luteal phases of the estrous cycle. The coEV were characterized by size distribution, polydispersity index (PDI), and zeta potential using dynamic light scattering (DLS) analysis, as well as, by transmission electron microscopy (TEM) and dot blotting (DB). Our results indicated that the physicochemical characteristics of the coEV were similar (P > 0.05), regardless of the isolation method (individual or pool). However, coEV collected during the luteal phase were larger (P < 0.05) than those during the follicular phase. The TEM showed spherical and cup-shaped particles, characteristic of exosomes. The DB revealed the presence of exosomal proteins involved in the biogenesis of coEV. In conclusion, it is possible to isolate and characterize coEV from a single caprine female and the estrous cycle phase influences the vesicles average size and PDI.

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来源期刊
Animal Reproduction
Animal Reproduction AGRICULTURE, DAIRY & ANIMAL SCIENCE-
CiteScore
2.30
自引率
11.80%
发文量
49
审稿时长
70 days
期刊介绍: Animal Reproduction (AR) publishes original scientific papers and invited literature reviews, in the form of Basic Research, Biotechnology, Applied Research and Review Articles, with the goal of contributing to a better understanding of phenomena related to animal reproduction. The scope of the journal applies to students, researchers and practitioners in the fields of veterinary, biology and animal science, also being of interest to practitioners of human medicine. Animal Reproduction Journal is the official organ of the Brazilian College of Animal Reproduction in Brazil.
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