使用猪卵泡液衍生的外泌体进行体外成熟,以替代传统方法。

IF 2.4 2区 农林科学 Q3 REPRODUCTIVE BIOLOGY
{"title":"使用猪卵泡液衍生的外泌体进行体外成熟,以替代传统方法。","authors":"","doi":"10.1016/j.theriogenology.2024.08.030","DOIUrl":null,"url":null,"abstract":"<div><p>Extracellular vesicles, also known as exosomes, influence numerous cellular functions by regulating different signaling pathways. However, their role in animal reproduction remains understudied. This study aimed to evaluate the effects of porcine follicular fluid-derived exosomes (pff-Exos) on porcine oocyte <em>in vitro</em> maturation and parthenogenetic embryo development. We obtained pff-Exos through mixed-method ultracentrifugation and size-exclusion chromatography. Transmission electron microscopy revealed an increase in the expression of exosome markers in the first four of thirteen fractions. The number of pff-Exo was 2.2 × 10<sup>6</sup> particles per microliter. The highest maturation rate of porcine oocytes treated with pff-Exo was observed with 1.1 × 10<sup>7</sup> particles of pff-Exo in the absence of porcine follicular fluid (pFF) culture conditions. Moreover, increased expression of <em>Gdf9</em> and <em>Bmp15</em> was observed. The developmental rate was the highest upon treatment with 1.1 × 10<sup>7</sup> particles of pff-Exo, which increased the total cell number in blastocysts. Embryonic development to the 2-cell stage was similar between the control and pff-Exo groups; however, development to the 4-cell stage and blastocyst was significantly increased in the pff-Exo group (61.6 ± 6.08 % and 29.72 ± 1.41 %, respectively; <em>P</em> &lt; 0.05) compared with that in the control group (42.0 ± 5.19 % and 18.14 ± 1.78 %, respectively). The expression levels of <em>Oct4</em>, <em>Sox2</em>, <em>Bcl2</em>, <em>Elf4</em>, and Gcn5 significantly increased at the pff-Exo 2-cell stage, whereas those of <em>Bax, Hdac1, Hdac6</em>, and <em>Sirt6</em> decreased. Specifically, the Oct4, Sox2, Elf4, Gcn5, and Hdac6 levels remained stable in pff-Exo 4-cell embryos, whereas those of p53 and <em>Hat1</em> were reduced and increased, respectively. Treatment with pffExos significantly increased H3K9 and H3K14 acetylation levels. These results demonstrate that pff-Exo affects the <em>in vitro</em> maturation of porcine oocytes and early embryonic development by regulating gene expression.</p></div>","PeriodicalId":23131,"journal":{"name":"Theriogenology","volume":null,"pages":null},"PeriodicalIF":2.4000,"publicationDate":"2024-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0093691X24003583/pdfft?md5=3ec3871d35eae886aef5453074fef629&pid=1-s2.0-S0093691X24003583-main.pdf","citationCount":"0","resultStr":"{\"title\":\"In vitro maturation using porcine follicular fluid-derived exosomes as an alternative to the conventional method\",\"authors\":\"\",\"doi\":\"10.1016/j.theriogenology.2024.08.030\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Extracellular vesicles, also known as exosomes, influence numerous cellular functions by regulating different signaling pathways. However, their role in animal reproduction remains understudied. This study aimed to evaluate the effects of porcine follicular fluid-derived exosomes (pff-Exos) on porcine oocyte <em>in vitro</em> maturation and parthenogenetic embryo development. We obtained pff-Exos through mixed-method ultracentrifugation and size-exclusion chromatography. Transmission electron microscopy revealed an increase in the expression of exosome markers in the first four of thirteen fractions. The number of pff-Exo was 2.2 × 10<sup>6</sup> particles per microliter. The highest maturation rate of porcine oocytes treated with pff-Exo was observed with 1.1 × 10<sup>7</sup> particles of pff-Exo in the absence of porcine follicular fluid (pFF) culture conditions. Moreover, increased expression of <em>Gdf9</em> and <em>Bmp15</em> was observed. The developmental rate was the highest upon treatment with 1.1 × 10<sup>7</sup> particles of pff-Exo, which increased the total cell number in blastocysts. Embryonic development to the 2-cell stage was similar between the control and pff-Exo groups; however, development to the 4-cell stage and blastocyst was significantly increased in the pff-Exo group (61.6 ± 6.08 % and 29.72 ± 1.41 %, respectively; <em>P</em> &lt; 0.05) compared with that in the control group (42.0 ± 5.19 % and 18.14 ± 1.78 %, respectively). The expression levels of <em>Oct4</em>, <em>Sox2</em>, <em>Bcl2</em>, <em>Elf4</em>, and Gcn5 significantly increased at the pff-Exo 2-cell stage, whereas those of <em>Bax, Hdac1, Hdac6</em>, and <em>Sirt6</em> decreased. Specifically, the Oct4, Sox2, Elf4, Gcn5, and Hdac6 levels remained stable in pff-Exo 4-cell embryos, whereas those of p53 and <em>Hat1</em> were reduced and increased, respectively. Treatment with pffExos significantly increased H3K9 and H3K14 acetylation levels. These results demonstrate that pff-Exo affects the <em>in vitro</em> maturation of porcine oocytes and early embryonic development by regulating gene expression.</p></div>\",\"PeriodicalId\":23131,\"journal\":{\"name\":\"Theriogenology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":2.4000,\"publicationDate\":\"2024-08-29\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S0093691X24003583/pdfft?md5=3ec3871d35eae886aef5453074fef629&pid=1-s2.0-S0093691X24003583-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Theriogenology\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0093691X24003583\",\"RegionNum\":2,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"REPRODUCTIVE BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Theriogenology","FirstCategoryId":"97","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0093691X24003583","RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"REPRODUCTIVE BIOLOGY","Score":null,"Total":0}
引用次数: 0

摘要

细胞外囊泡,又称外泌体,通过调节不同的信号通路影响多种细胞功能。然而,它们在动物繁殖中的作用仍未得到充分研究。本研究旨在评估猪卵泡液衍生外泌体(pff-Exos)对猪卵母细胞体外成熟和孤雌胚发育的影响。我们通过混合超速离心和大小排阻色谱法获得了 pff-Exos。透射电子显微镜显示,在 13 个馏分的前 4 个中,外泌体标记物的表达量有所增加。pff-Exo 的数量为每微升 2.2 × 106 个。在无猪卵泡液(pFF)培养条件下,用 pff-Exo 处理的猪卵母细胞成熟率最高,达到 1.1 × 107 个 pff-Exo 颗粒。此外,还观察到 Gdf9 和 Bmp15 的表达增加。用 1.1 × 107 颗粒 pff-Exo 处理时,胚胎发育率最高,囊胚中的细胞总数也增加了。对照组和 pff-Exo 组的胚胎发育至 2 细胞期相似;但 pff-Exo 组的胚胎发育至 4 细胞期和囊胚期显著增加(分别为 61.6 ± 6.08 % 和 29.72 ± 1.41 %;P.
本文章由计算机程序翻译,如有差异,请以英文原文为准。
In vitro maturation using porcine follicular fluid-derived exosomes as an alternative to the conventional method

Extracellular vesicles, also known as exosomes, influence numerous cellular functions by regulating different signaling pathways. However, their role in animal reproduction remains understudied. This study aimed to evaluate the effects of porcine follicular fluid-derived exosomes (pff-Exos) on porcine oocyte in vitro maturation and parthenogenetic embryo development. We obtained pff-Exos through mixed-method ultracentrifugation and size-exclusion chromatography. Transmission electron microscopy revealed an increase in the expression of exosome markers in the first four of thirteen fractions. The number of pff-Exo was 2.2 × 106 particles per microliter. The highest maturation rate of porcine oocytes treated with pff-Exo was observed with 1.1 × 107 particles of pff-Exo in the absence of porcine follicular fluid (pFF) culture conditions. Moreover, increased expression of Gdf9 and Bmp15 was observed. The developmental rate was the highest upon treatment with 1.1 × 107 particles of pff-Exo, which increased the total cell number in blastocysts. Embryonic development to the 2-cell stage was similar between the control and pff-Exo groups; however, development to the 4-cell stage and blastocyst was significantly increased in the pff-Exo group (61.6 ± 6.08 % and 29.72 ± 1.41 %, respectively; P < 0.05) compared with that in the control group (42.0 ± 5.19 % and 18.14 ± 1.78 %, respectively). The expression levels of Oct4, Sox2, Bcl2, Elf4, and Gcn5 significantly increased at the pff-Exo 2-cell stage, whereas those of Bax, Hdac1, Hdac6, and Sirt6 decreased. Specifically, the Oct4, Sox2, Elf4, Gcn5, and Hdac6 levels remained stable in pff-Exo 4-cell embryos, whereas those of p53 and Hat1 were reduced and increased, respectively. Treatment with pffExos significantly increased H3K9 and H3K14 acetylation levels. These results demonstrate that pff-Exo affects the in vitro maturation of porcine oocytes and early embryonic development by regulating gene expression.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
Theriogenology
Theriogenology 农林科学-生殖生物学
CiteScore
5.50
自引率
14.30%
发文量
387
审稿时长
72 days
期刊介绍: Theriogenology provides an international forum for researchers, clinicians, and industry professionals in animal reproductive biology. This acclaimed journal publishes articles on a wide range of topics in reproductive and developmental biology, of domestic mammal, avian, and aquatic species as well as wild species which are the object of veterinary care in research or conservation programs.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信