精子衍生的中心粒是哺乳动物生殖细胞有丝分裂初期染色体准确分离的重要贡献者

IF 3.7 2区 医学 Q1 OBSTETRICS & GYNECOLOGY Reproductive biomedicine online Pub Date : 2024-05-01 DOI:10.1016/j.rbmo.2024.104026
A. Larreategui Aparicio , I. Barale , C. Deelen , T.A. Stout , G.J. Kops , M. de Ruijter-Villani
{"title":"精子衍生的中心粒是哺乳动物生殖细胞有丝分裂初期染色体准确分离的重要贡献者","authors":"A. Larreategui Aparicio ,&nbsp;I. Barale ,&nbsp;C. Deelen ,&nbsp;T.A. Stout ,&nbsp;G.J. Kops ,&nbsp;M. de Ruijter-Villani","doi":"10.1016/j.rbmo.2024.104026","DOIUrl":null,"url":null,"abstract":"<div><h3>BACKGROUND AND AIM</h3><p>The centrosomes are the major microtubule-organizing centers in somatic cells and have an essential role in spindle assembly and chromosome segregation. Centrioles are also found in the zygotes, but they are introduced by the spermatozoon at fertilization and they are structurally different to the canonical centrioles. We recently demonstrated that centrosomes play a minor role during zygotic spindle assembly, but its role in chromosome segregation remains unknown. The aim of this study is to investigate if centrosome malpositioning or inactivity predisposes to chromosome segregation errors and failure of cytokinesis.</p></div><div><h3>METHODS</h3><p>Bovine oocytes were retrieved from slaughterhouse ovaries and matured in vitro. Matured oocytes were fertilized in vitro with semen from a bull of proven fertility and, either fixed at 27-29h post-fertilization, or used for time-lapse light-sheet imaging after injection of mRNA for microtubule and chromatin reporters. Fixed bovine zygotes were immunostained for chromatin, alpha-tubulin, centriolar and pericentriolar components, and imaged with a confocal microscope. Microtubule nucleation ability was evaluated using a microtubule regrowth assay.</p></div><div><h3>RESULTS</h3><p>Live imaging showed that in 52% of bovine zygotes (24/46) the centrosomes engaged with the spindle but were malpositioned, while in 26% (12/46) the centrosomes were not visible (inactive). All zygotes with normally positioned centrosomes showed correct chromosome segregation, whereas 44% of zygotes with abnormal centriole positioning showed lagging chromosomes and the formation of micronuclei.</p><p>When centriole duplication was prevented, the sperm-derived centrioles showed a similar ability to recruit pericentrosomal material (NEDD1) and nucleate microtubules. However, live imaging showed that in 82% (14/17) of PLK4i treated zygotes the spindle, although bipolar, was unusually elongated and chromosomes either failed to align correctly on the metaphase plate or showed anaphase lag. Moreover, in treated zygotes where both centrioles were positioned at the same spindle pole, the spindle failed to anchor correctly to the cell membrane, resulting in rotational behavior of the zygote and an increase in cytokinesis failure of up to 47% (8/17).</p></div><div><h3>CONCLUSIONS</h3><p>These results show how correct centrosome positioning guarantees spindle stability and correct chromosome segregation. On the other hand, when centrosomes are malpositioned or inactive, there is an increase in chromosome segregation errors and failure of cytokinesis.</p></div>","PeriodicalId":21134,"journal":{"name":"Reproductive biomedicine online","volume":null,"pages":null},"PeriodicalIF":3.7000,"publicationDate":"2024-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Sperm-derived Centrioles are Essential Contributors to Accurate Chromosome Segregation in the Initial Mitotic Division of Mammalian Zygotes\",\"authors\":\"A. Larreategui Aparicio ,&nbsp;I. Barale ,&nbsp;C. Deelen ,&nbsp;T.A. Stout ,&nbsp;G.J. Kops ,&nbsp;M. de Ruijter-Villani\",\"doi\":\"10.1016/j.rbmo.2024.104026\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>BACKGROUND AND AIM</h3><p>The centrosomes are the major microtubule-organizing centers in somatic cells and have an essential role in spindle assembly and chromosome segregation. Centrioles are also found in the zygotes, but they are introduced by the spermatozoon at fertilization and they are structurally different to the canonical centrioles. We recently demonstrated that centrosomes play a minor role during zygotic spindle assembly, but its role in chromosome segregation remains unknown. The aim of this study is to investigate if centrosome malpositioning or inactivity predisposes to chromosome segregation errors and failure of cytokinesis.</p></div><div><h3>METHODS</h3><p>Bovine oocytes were retrieved from slaughterhouse ovaries and matured in vitro. Matured oocytes were fertilized in vitro with semen from a bull of proven fertility and, either fixed at 27-29h post-fertilization, or used for time-lapse light-sheet imaging after injection of mRNA for microtubule and chromatin reporters. Fixed bovine zygotes were immunostained for chromatin, alpha-tubulin, centriolar and pericentriolar components, and imaged with a confocal microscope. Microtubule nucleation ability was evaluated using a microtubule regrowth assay.</p></div><div><h3>RESULTS</h3><p>Live imaging showed that in 52% of bovine zygotes (24/46) the centrosomes engaged with the spindle but were malpositioned, while in 26% (12/46) the centrosomes were not visible (inactive). All zygotes with normally positioned centrosomes showed correct chromosome segregation, whereas 44% of zygotes with abnormal centriole positioning showed lagging chromosomes and the formation of micronuclei.</p><p>When centriole duplication was prevented, the sperm-derived centrioles showed a similar ability to recruit pericentrosomal material (NEDD1) and nucleate microtubules. However, live imaging showed that in 82% (14/17) of PLK4i treated zygotes the spindle, although bipolar, was unusually elongated and chromosomes either failed to align correctly on the metaphase plate or showed anaphase lag. Moreover, in treated zygotes where both centrioles were positioned at the same spindle pole, the spindle failed to anchor correctly to the cell membrane, resulting in rotational behavior of the zygote and an increase in cytokinesis failure of up to 47% (8/17).</p></div><div><h3>CONCLUSIONS</h3><p>These results show how correct centrosome positioning guarantees spindle stability and correct chromosome segregation. On the other hand, when centrosomes are malpositioned or inactive, there is an increase in chromosome segregation errors and failure of cytokinesis.</p></div>\",\"PeriodicalId\":21134,\"journal\":{\"name\":\"Reproductive biomedicine online\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":3.7000,\"publicationDate\":\"2024-05-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Reproductive biomedicine online\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1472648324002153\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"OBSTETRICS & GYNECOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Reproductive biomedicine online","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1472648324002153","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"OBSTETRICS & GYNECOLOGY","Score":null,"Total":0}
引用次数: 0

摘要

背景和目的中心体是体细胞中主要的微管组织中心,在纺锤体组装和染色体分离中起着至关重要的作用。中心体也存在于子代细胞中,但它们是由精子在受精时引入的,在结构上与典型的中心体不同。我们最近证明,中心粒在胚胎纺锤体组装过程中起着次要作用,但它在染色体分离中的作用仍然未知。本研究旨在探讨中心体定位错误或不活跃是否容易导致染色体分离错误和细胞分裂失败。成熟的卵母细胞与经证实具有生育能力的公牛精液体外受精,并在受精后 27-29 小时进行固定,或在注入微管和染色质报告的 mRNA 后用于延时光片成像。对固定的牛胚胎进行染色质、α-微管蛋白、中心粒和周粒成分的免疫染色,并用共聚焦显微镜成像。结果实时成像显示,52%(24/46)的牛胚胎中心体与纺锤体接触但位置不正,而 26%(12/46)的牛胚胎中心体不可见(不活跃)。中心粒定位正常的所有胚胎都表现出正确的染色体分离,而中心粒定位异常的44%的胚胎则表现出染色体滞后和微核的形成。然而,活体成像显示,在82%(14/17)的PLK4i处理过的子实体中,纺锤体虽然是双极的,但却异常拉长,染色体要么不能在分裂板上正确排列,要么出现无丝分裂滞后。此外,在两个中心粒都位于同一纺锤极的处理后子中,纺锤体未能正确锚定到细胞膜上,导致后子发生旋转,细胞分裂失败率增加高达 47% (8/17)。另一方面,当中心体定位错误或不活跃时,染色体分离错误和细胞分裂失败的情况会增加。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
Sperm-derived Centrioles are Essential Contributors to Accurate Chromosome Segregation in the Initial Mitotic Division of Mammalian Zygotes

BACKGROUND AND AIM

The centrosomes are the major microtubule-organizing centers in somatic cells and have an essential role in spindle assembly and chromosome segregation. Centrioles are also found in the zygotes, but they are introduced by the spermatozoon at fertilization and they are structurally different to the canonical centrioles. We recently demonstrated that centrosomes play a minor role during zygotic spindle assembly, but its role in chromosome segregation remains unknown. The aim of this study is to investigate if centrosome malpositioning or inactivity predisposes to chromosome segregation errors and failure of cytokinesis.

METHODS

Bovine oocytes were retrieved from slaughterhouse ovaries and matured in vitro. Matured oocytes were fertilized in vitro with semen from a bull of proven fertility and, either fixed at 27-29h post-fertilization, or used for time-lapse light-sheet imaging after injection of mRNA for microtubule and chromatin reporters. Fixed bovine zygotes were immunostained for chromatin, alpha-tubulin, centriolar and pericentriolar components, and imaged with a confocal microscope. Microtubule nucleation ability was evaluated using a microtubule regrowth assay.

RESULTS

Live imaging showed that in 52% of bovine zygotes (24/46) the centrosomes engaged with the spindle but were malpositioned, while in 26% (12/46) the centrosomes were not visible (inactive). All zygotes with normally positioned centrosomes showed correct chromosome segregation, whereas 44% of zygotes with abnormal centriole positioning showed lagging chromosomes and the formation of micronuclei.

When centriole duplication was prevented, the sperm-derived centrioles showed a similar ability to recruit pericentrosomal material (NEDD1) and nucleate microtubules. However, live imaging showed that in 82% (14/17) of PLK4i treated zygotes the spindle, although bipolar, was unusually elongated and chromosomes either failed to align correctly on the metaphase plate or showed anaphase lag. Moreover, in treated zygotes where both centrioles were positioned at the same spindle pole, the spindle failed to anchor correctly to the cell membrane, resulting in rotational behavior of the zygote and an increase in cytokinesis failure of up to 47% (8/17).

CONCLUSIONS

These results show how correct centrosome positioning guarantees spindle stability and correct chromosome segregation. On the other hand, when centrosomes are malpositioned or inactive, there is an increase in chromosome segregation errors and failure of cytokinesis.

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
Reproductive biomedicine online
Reproductive biomedicine online 医学-妇产科学
CiteScore
7.20
自引率
7.50%
发文量
391
审稿时长
50 days
期刊介绍: Reproductive BioMedicine Online covers the formation, growth and differentiation of the human embryo. It is intended to bring to public attention new research on biological and clinical research on human reproduction and the human embryo including relevant studies on animals. It is published by a group of scientists and clinicians working in these fields of study. Its audience comprises researchers, clinicians, practitioners, academics and patients. Context: The period of human embryonic growth covered is between the formation of the primordial germ cells in the fetus until mid-pregnancy. High quality research on lower animals is included if it helps to clarify the human situation. Studies progressing to birth and later are published if they have a direct bearing on events in the earlier stages of pregnancy.
期刊最新文献
Ultra-fast vitrification and rapid elution of human oocytes: part I. germinal vesicle model validation. Ultra-fast vitrification and rapid elution of human oocytes: Part II - verification of blastocyst development from mature oocytes. Inside Front Cover - Affiliations and First page of TOC Front Matter - Continued TOC Outside Back Cover - Editorial Board
×
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
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 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 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1