Mendes Roberto, de Alcântara Agostinho Soares, Tetaping Gildas Mbemya, Novaes Marco Aurélio Schiavo, Gonçalves Vanessa Barbosa Pinheiro, da Silva João Xavier, de Almeida José Jonathas Albuquerque, Guedes Maria Izabel Florindo, Leal Luzia Kalyne Almeida Moreira, Souza-Fabjan Joanna Maria Gonçalves, Nicolete Roberto, Padilha Deborah de Melo Magalhães, de Figueiredo José Ricardo, Rodrigues Ana Paula Ribeiro
Laboratório de Manipulação de Oócitos e Folículos Ovarianos Pré-antrais - LAMOFOPA, Faculdade de Medicina Veterinária, Universidade Estadual do Ceará - UECE, Fortaleza, CE, Brasil.
Faculdade de Educação e Ciências Integradas de Crateús, Universidade Estadual do Ceará - UECE, Fortaleza, CE, Brasil.
Anim Reprod. 2024 Oct 21;21(4):e20240039. doi: 10.1590/1984-3143-AR2024-0039. eCollection 2024.
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.
输卵管液细胞外囊泡（oEV）对于围受孕事件至关重要。除了山羊外，在哺乳动物的输卵管液（OF）中已鉴定出EV的存在。因此，本研究旨在分离并表征山羊oEV（coEV）。最初，在实验1中，分别从每只动物或三只动物的混合样本的OF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。使用动态光散射（DLS）分析，以及通过透射电子显微镜（TEM）和斑点印迹（DB）对coEV进行大小分布、多分散指数（PDI）和zeta电位表征。我们的结果表明，无论分离方法是单独分离还是混合分离，coEV的物理化学特性相似（P>0.05）。然而，在黄体期收集的coEV比卵泡期的更大（P<0.05）。TEM显示出球形和杯形颗粒，这是外泌体的特征。DB揭示了参与coEV生物发生的外泌体蛋白的存在。总之，有可能从单个山羊雌性个体中分离并表征coEV，并且发情周期阶段会影响囊泡的平均大小和PDI。
