Cargando…

Integrated transcriptomics and proteomics assay identifies the role of FCGR1A in maintaining sperm fertilization capacity during semen cryopreservation in sheep

Semen cryopreservation is a promising technology employed in preserving high-quality varieties in animal husbandry and is also widely applied in the human sperm bank. However, the compromised qualities, such as decreased sperm motility, damaged membrane structure, and reduced fertilization competenc...

Descripción completa

Detalles Bibliográficos
Autores principales: Bai, Jiachen, Zhou, Guizhen, Hao, Shaopeng, Liu, Yucheng, Guo, Yanhua, Wang, Jingjing, Liu, Hongtao, Wang, Longfei, Li, Jun, Liu, Aiju, Sun, Wendell Q., Wan, Pengcheng, Fu, Xiangwei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10433746/
https://www.ncbi.nlm.nih.gov/pubmed/37601105
http://dx.doi.org/10.3389/fcell.2023.1177774
_version_ 1785091717686362112
author Bai, Jiachen
Zhou, Guizhen
Hao, Shaopeng
Liu, Yucheng
Guo, Yanhua
Wang, Jingjing
Liu, Hongtao
Wang, Longfei
Li, Jun
Liu, Aiju
Sun, Wendell Q.
Wan, Pengcheng
Fu, Xiangwei
author_facet Bai, Jiachen
Zhou, Guizhen
Hao, Shaopeng
Liu, Yucheng
Guo, Yanhua
Wang, Jingjing
Liu, Hongtao
Wang, Longfei
Li, Jun
Liu, Aiju
Sun, Wendell Q.
Wan, Pengcheng
Fu, Xiangwei
author_sort Bai, Jiachen
collection PubMed
description Semen cryopreservation is a promising technology employed in preserving high-quality varieties in animal husbandry and is also widely applied in the human sperm bank. However, the compromised qualities, such as decreased sperm motility, damaged membrane structure, and reduced fertilization competency, have significantly hampered the efficient application of this technique. Therefore, it is imperative to depict various molecular changes found in cryopreserved sperm and identify the regulatory network in response to the cryopreservation stress. In this study, semen was collected from three Chinese Merino rams and divided into untreated (fresh semen, FS) and programmed freezing (programmed freezing semen, PS) groups. After measuring different quality parameters, the ultra-low RNA-seq and tandem mass tag-based (TMT) proteome were conducted in both the groups. The results indicated that the motility (82.63% ± 3.55% vs. 34.10% ± 2.90%, p < 0.05) and viability (89.46% ± 2.53% vs. 44.78% ± 2.29%, p < 0.05) of the sperm in the FS group were significantly higher compared to those in the PS group. In addition, 45 upregulated and 291 downregulated genes, as well as 30 upregulated and 48 downregulated proteins, were found in transcriptomics and proteomics data separately. Moreover, three integrated methods, namely, functional annotation and enrichment analysis, Pearson’s correlation analysis, and two-way orthogonal partial least squares (O2PLS) analysis, were used for further analysis. The results suggested that various differentially expressed genes and proteins (DEGs and DEPs) were mainly enriched in leishmaniasis and hematopoietic cell lineage, and Fc gamma receptor Ia (FCGR1A) was significantly downregulated in cryopreserved sperm both at mRNA and protein levels in comparison with the fresh counterpart. In addition, top five genes (FCGR1A, HCK, SLX4, ITGA3, and BET1) and 22 proteins could form a distinct network in which genes and proteins were significantly correlated (p < 0.05). Interestingly, FCGR1A also appeared in the top 25 correlation list based on O2PLS analysis. Hence, FCGR1A was selected as the most potential differentially expressed candidate for screening by the three integrated multi-omics analysis methods. In addition, Pearson’s correlation analysis indicated that the expression level of FCGR1A was positively correlated with sperm motility and viability. A subsequent experiment was conducted to identify the biological role of FCGR1A in sperm function. The results showed that both the sperm viability (fresh group: 87.65% ± 4.17% vs. 75.8% ± 1.15%, cryopreserved group: 48.15% ± 0.63% vs. 42.45% ± 2.61%, p < 0.05) and motility (fresh group: 83.27% ± 4.15% vs. 70.41% ± 1.07%, cryopreserved group: 45.31% ± 3.28% vs. 35.13% ± 2.82%, p < 0.05) were significantly reduced in fresh and frozen sperm when FCGR1A was blocked. Moreover, the cleavage rate of embryos fertilized by FCGR1A-blocked sperm was noted to be significantly lower in both fresh (95.28% ± 1.16% vs. 90.44% ± 1.56%, p < 0.05) and frozen groups (89.8% ± 1.50% vs. 82.53% ± 1.53%, p < 0.05). In conclusion, our results revealed that the downregulated membrane protein FCGR1A can potentially contribute to the reduced sperm fertility competency in the cryopreserved sheep sperm.
format Online
Article
Text
id pubmed-10433746
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-104337462023-08-18 Integrated transcriptomics and proteomics assay identifies the role of FCGR1A in maintaining sperm fertilization capacity during semen cryopreservation in sheep Bai, Jiachen Zhou, Guizhen Hao, Shaopeng Liu, Yucheng Guo, Yanhua Wang, Jingjing Liu, Hongtao Wang, Longfei Li, Jun Liu, Aiju Sun, Wendell Q. Wan, Pengcheng Fu, Xiangwei Front Cell Dev Biol Cell and Developmental Biology Semen cryopreservation is a promising technology employed in preserving high-quality varieties in animal husbandry and is also widely applied in the human sperm bank. However, the compromised qualities, such as decreased sperm motility, damaged membrane structure, and reduced fertilization competency, have significantly hampered the efficient application of this technique. Therefore, it is imperative to depict various molecular changes found in cryopreserved sperm and identify the regulatory network in response to the cryopreservation stress. In this study, semen was collected from three Chinese Merino rams and divided into untreated (fresh semen, FS) and programmed freezing (programmed freezing semen, PS) groups. After measuring different quality parameters, the ultra-low RNA-seq and tandem mass tag-based (TMT) proteome were conducted in both the groups. The results indicated that the motility (82.63% ± 3.55% vs. 34.10% ± 2.90%, p < 0.05) and viability (89.46% ± 2.53% vs. 44.78% ± 2.29%, p < 0.05) of the sperm in the FS group were significantly higher compared to those in the PS group. In addition, 45 upregulated and 291 downregulated genes, as well as 30 upregulated and 48 downregulated proteins, were found in transcriptomics and proteomics data separately. Moreover, three integrated methods, namely, functional annotation and enrichment analysis, Pearson’s correlation analysis, and two-way orthogonal partial least squares (O2PLS) analysis, were used for further analysis. The results suggested that various differentially expressed genes and proteins (DEGs and DEPs) were mainly enriched in leishmaniasis and hematopoietic cell lineage, and Fc gamma receptor Ia (FCGR1A) was significantly downregulated in cryopreserved sperm both at mRNA and protein levels in comparison with the fresh counterpart. In addition, top five genes (FCGR1A, HCK, SLX4, ITGA3, and BET1) and 22 proteins could form a distinct network in which genes and proteins were significantly correlated (p < 0.05). Interestingly, FCGR1A also appeared in the top 25 correlation list based on O2PLS analysis. Hence, FCGR1A was selected as the most potential differentially expressed candidate for screening by the three integrated multi-omics analysis methods. In addition, Pearson’s correlation analysis indicated that the expression level of FCGR1A was positively correlated with sperm motility and viability. A subsequent experiment was conducted to identify the biological role of FCGR1A in sperm function. The results showed that both the sperm viability (fresh group: 87.65% ± 4.17% vs. 75.8% ± 1.15%, cryopreserved group: 48.15% ± 0.63% vs. 42.45% ± 2.61%, p < 0.05) and motility (fresh group: 83.27% ± 4.15% vs. 70.41% ± 1.07%, cryopreserved group: 45.31% ± 3.28% vs. 35.13% ± 2.82%, p < 0.05) were significantly reduced in fresh and frozen sperm when FCGR1A was blocked. Moreover, the cleavage rate of embryos fertilized by FCGR1A-blocked sperm was noted to be significantly lower in both fresh (95.28% ± 1.16% vs. 90.44% ± 1.56%, p < 0.05) and frozen groups (89.8% ± 1.50% vs. 82.53% ± 1.53%, p < 0.05). In conclusion, our results revealed that the downregulated membrane protein FCGR1A can potentially contribute to the reduced sperm fertility competency in the cryopreserved sheep sperm. Frontiers Media S.A. 2023-08-02 /pmc/articles/PMC10433746/ /pubmed/37601105 http://dx.doi.org/10.3389/fcell.2023.1177774 Text en Copyright © 2023 Bai, Zhou, Hao, Liu, Guo, Wang, Liu, Wang, Li, Liu, Sun, Wan and Fu. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cell and Developmental Biology
Bai, Jiachen
Zhou, Guizhen
Hao, Shaopeng
Liu, Yucheng
Guo, Yanhua
Wang, Jingjing
Liu, Hongtao
Wang, Longfei
Li, Jun
Liu, Aiju
Sun, Wendell Q.
Wan, Pengcheng
Fu, Xiangwei
Integrated transcriptomics and proteomics assay identifies the role of FCGR1A in maintaining sperm fertilization capacity during semen cryopreservation in sheep
title Integrated transcriptomics and proteomics assay identifies the role of FCGR1A in maintaining sperm fertilization capacity during semen cryopreservation in sheep
title_full Integrated transcriptomics and proteomics assay identifies the role of FCGR1A in maintaining sperm fertilization capacity during semen cryopreservation in sheep
title_fullStr Integrated transcriptomics and proteomics assay identifies the role of FCGR1A in maintaining sperm fertilization capacity during semen cryopreservation in sheep
title_full_unstemmed Integrated transcriptomics and proteomics assay identifies the role of FCGR1A in maintaining sperm fertilization capacity during semen cryopreservation in sheep
title_short Integrated transcriptomics and proteomics assay identifies the role of FCGR1A in maintaining sperm fertilization capacity during semen cryopreservation in sheep
title_sort integrated transcriptomics and proteomics assay identifies the role of fcgr1a in maintaining sperm fertilization capacity during semen cryopreservation in sheep
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10433746/
https://www.ncbi.nlm.nih.gov/pubmed/37601105
http://dx.doi.org/10.3389/fcell.2023.1177774
work_keys_str_mv AT baijiachen integratedtranscriptomicsandproteomicsassayidentifiestheroleoffcgr1ainmaintainingspermfertilizationcapacityduringsemencryopreservationinsheep
AT zhouguizhen integratedtranscriptomicsandproteomicsassayidentifiestheroleoffcgr1ainmaintainingspermfertilizationcapacityduringsemencryopreservationinsheep
AT haoshaopeng integratedtranscriptomicsandproteomicsassayidentifiestheroleoffcgr1ainmaintainingspermfertilizationcapacityduringsemencryopreservationinsheep
AT liuyucheng integratedtranscriptomicsandproteomicsassayidentifiestheroleoffcgr1ainmaintainingspermfertilizationcapacityduringsemencryopreservationinsheep
AT guoyanhua integratedtranscriptomicsandproteomicsassayidentifiestheroleoffcgr1ainmaintainingspermfertilizationcapacityduringsemencryopreservationinsheep
AT wangjingjing integratedtranscriptomicsandproteomicsassayidentifiestheroleoffcgr1ainmaintainingspermfertilizationcapacityduringsemencryopreservationinsheep
AT liuhongtao integratedtranscriptomicsandproteomicsassayidentifiestheroleoffcgr1ainmaintainingspermfertilizationcapacityduringsemencryopreservationinsheep
AT wanglongfei integratedtranscriptomicsandproteomicsassayidentifiestheroleoffcgr1ainmaintainingspermfertilizationcapacityduringsemencryopreservationinsheep
AT lijun integratedtranscriptomicsandproteomicsassayidentifiestheroleoffcgr1ainmaintainingspermfertilizationcapacityduringsemencryopreservationinsheep
AT liuaiju integratedtranscriptomicsandproteomicsassayidentifiestheroleoffcgr1ainmaintainingspermfertilizationcapacityduringsemencryopreservationinsheep
AT sunwendellq integratedtranscriptomicsandproteomicsassayidentifiestheroleoffcgr1ainmaintainingspermfertilizationcapacityduringsemencryopreservationinsheep
AT wanpengcheng integratedtranscriptomicsandproteomicsassayidentifiestheroleoffcgr1ainmaintainingspermfertilizationcapacityduringsemencryopreservationinsheep
AT fuxiangwei integratedtranscriptomicsandproteomicsassayidentifiestheroleoffcgr1ainmaintainingspermfertilizationcapacityduringsemencryopreservationinsheep