Cargando…

Influence of sire fertility status on conceptus-induced transcriptomic response of the bovine endometrium

The aim was to examine the effect of sire fertility status on conceptus-induced changes in the bovine endometrial transcriptome. To generate elongated conceptuses, Day 7 blastocysts produced in vitro using frozen-thawed sperm from Holstein Friesian bulls (3 High fertility, HF and 3 Low fertility, LF...

Descripción completa

Detalles Bibliográficos
Autores principales: O’Callaghan, E., Sánchez, J.M., Rabaglino, M.B., McDonald, M., Liu, H., Spencer, T.E., Fair, S., Kenny, D.A., Lonergan, P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9442353/
https://www.ncbi.nlm.nih.gov/pubmed/36072344
http://dx.doi.org/10.3389/fcell.2022.950443
_version_ 1784782794771136512
author O’Callaghan, E.
Sánchez, J.M.
Rabaglino, M.B.
McDonald, M.
Liu, H.
Spencer, T.E.
Fair, S.
Kenny, D.A.
Lonergan, P.
author_facet O’Callaghan, E.
Sánchez, J.M.
Rabaglino, M.B.
McDonald, M.
Liu, H.
Spencer, T.E.
Fair, S.
Kenny, D.A.
Lonergan, P.
author_sort O’Callaghan, E.
collection PubMed
description The aim was to examine the effect of sire fertility status on conceptus-induced changes in the bovine endometrial transcriptome. To generate elongated conceptuses, Day 7 blastocysts produced in vitro using frozen-thawed sperm from Holstein Friesian bulls (3 High fertility, HF and 3 Low fertility, LF) were transferred in groups of 5–10 into synchronized heifers (n = 7 heifers per bull) and recovered following slaughter on Day 15. Day 15 endometrial explants recovered from the uterine horn ipsilateral to the corpus luteum were recovered from synchronized cyclic heifers (n = 4). Explants from each heifer were co-cultured for 6 h in RPMI medium alone (Control) or with 100 ng/ml ovine recombinant interferon tau (IFNT) or with a single conceptus from each HF or LF bull. After 6 h, explants were snap frozen and stored at −80°C. Extracted mRNA was subjected to RNA-seq and the resulting data were analyzed with R software. The numbers of differentially expressed genes (DEG; FDR<0.05) were: HF vs. Control: 956; LF vs. Control: 1021; IFNT vs. Control: 1301; HF vs. LF: 2. Unsurprisingly, the majority of DEG (658) were common to all comparisons and were related to IFNT-induced changes in the endometrium. Prior to applying the adjusted p-value, there were 700 DEG between HF and LF, with 191 and 509 genes more expressed in HF or LF, respectively (p < 0.05). Overrepresentation analysis of KEGG pathways (FDR<0.05), revealed that DEG with higher expression in LF were involved in cell cycle and proteolysis, while those upregulated DEG by HF conceptuses were strongly associated with immune process pathways, such as TNF, NF-kappa B, cytokine-cytokine receptor interaction, and TLR signaling. These pathways were also enriched by DEG upregulated by IFNT compared to the Control. Furthermore, only the HF, and not the LF group, affected the expression of most genes in these pathways (p < 0.05) according to a negative binomial regression model. Finally, a weighted gene co-expression network analysis revealed two clusters of co-expressed genes associated with the HF conceptuses (p < 0.05), which were also enriched for the aforementioned pathways. In conclusion, HF conceptuses, similar to IFNT treatment, stimulated multiple pathways involved in immune response, which were apparently not affected by LF conceptuses.
format Online
Article
Text
id pubmed-9442353
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-94423532022-09-06 Influence of sire fertility status on conceptus-induced transcriptomic response of the bovine endometrium O’Callaghan, E. Sánchez, J.M. Rabaglino, M.B. McDonald, M. Liu, H. Spencer, T.E. Fair, S. Kenny, D.A. Lonergan, P. Front Cell Dev Biol Cell and Developmental Biology The aim was to examine the effect of sire fertility status on conceptus-induced changes in the bovine endometrial transcriptome. To generate elongated conceptuses, Day 7 blastocysts produced in vitro using frozen-thawed sperm from Holstein Friesian bulls (3 High fertility, HF and 3 Low fertility, LF) were transferred in groups of 5–10 into synchronized heifers (n = 7 heifers per bull) and recovered following slaughter on Day 15. Day 15 endometrial explants recovered from the uterine horn ipsilateral to the corpus luteum were recovered from synchronized cyclic heifers (n = 4). Explants from each heifer were co-cultured for 6 h in RPMI medium alone (Control) or with 100 ng/ml ovine recombinant interferon tau (IFNT) or with a single conceptus from each HF or LF bull. After 6 h, explants were snap frozen and stored at −80°C. Extracted mRNA was subjected to RNA-seq and the resulting data were analyzed with R software. The numbers of differentially expressed genes (DEG; FDR<0.05) were: HF vs. Control: 956; LF vs. Control: 1021; IFNT vs. Control: 1301; HF vs. LF: 2. Unsurprisingly, the majority of DEG (658) were common to all comparisons and were related to IFNT-induced changes in the endometrium. Prior to applying the adjusted p-value, there were 700 DEG between HF and LF, with 191 and 509 genes more expressed in HF or LF, respectively (p < 0.05). Overrepresentation analysis of KEGG pathways (FDR<0.05), revealed that DEG with higher expression in LF were involved in cell cycle and proteolysis, while those upregulated DEG by HF conceptuses were strongly associated with immune process pathways, such as TNF, NF-kappa B, cytokine-cytokine receptor interaction, and TLR signaling. These pathways were also enriched by DEG upregulated by IFNT compared to the Control. Furthermore, only the HF, and not the LF group, affected the expression of most genes in these pathways (p < 0.05) according to a negative binomial regression model. Finally, a weighted gene co-expression network analysis revealed two clusters of co-expressed genes associated with the HF conceptuses (p < 0.05), which were also enriched for the aforementioned pathways. In conclusion, HF conceptuses, similar to IFNT treatment, stimulated multiple pathways involved in immune response, which were apparently not affected by LF conceptuses. Frontiers Media S.A. 2022-08-22 /pmc/articles/PMC9442353/ /pubmed/36072344 http://dx.doi.org/10.3389/fcell.2022.950443 Text en Copyright © 2022 O’Callaghan, Sánchez, Rabaglino, McDonald, Liu, Spencer, Fair, Kenny and Lonergan. 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
O’Callaghan, E.
Sánchez, J.M.
Rabaglino, M.B.
McDonald, M.
Liu, H.
Spencer, T.E.
Fair, S.
Kenny, D.A.
Lonergan, P.
Influence of sire fertility status on conceptus-induced transcriptomic response of the bovine endometrium
title Influence of sire fertility status on conceptus-induced transcriptomic response of the bovine endometrium
title_full Influence of sire fertility status on conceptus-induced transcriptomic response of the bovine endometrium
title_fullStr Influence of sire fertility status on conceptus-induced transcriptomic response of the bovine endometrium
title_full_unstemmed Influence of sire fertility status on conceptus-induced transcriptomic response of the bovine endometrium
title_short Influence of sire fertility status on conceptus-induced transcriptomic response of the bovine endometrium
title_sort influence of sire fertility status on conceptus-induced transcriptomic response of the bovine endometrium
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9442353/
https://www.ncbi.nlm.nih.gov/pubmed/36072344
http://dx.doi.org/10.3389/fcell.2022.950443
work_keys_str_mv AT ocallaghane influenceofsirefertilitystatusonconceptusinducedtranscriptomicresponseofthebovineendometrium
AT sanchezjm influenceofsirefertilitystatusonconceptusinducedtranscriptomicresponseofthebovineendometrium
AT rabaglinomb influenceofsirefertilitystatusonconceptusinducedtranscriptomicresponseofthebovineendometrium
AT mcdonaldm influenceofsirefertilitystatusonconceptusinducedtranscriptomicresponseofthebovineendometrium
AT liuh influenceofsirefertilitystatusonconceptusinducedtranscriptomicresponseofthebovineendometrium
AT spencerte influenceofsirefertilitystatusonconceptusinducedtranscriptomicresponseofthebovineendometrium
AT fairs influenceofsirefertilitystatusonconceptusinducedtranscriptomicresponseofthebovineendometrium
AT kennyda influenceofsirefertilitystatusonconceptusinducedtranscriptomicresponseofthebovineendometrium
AT lonerganp influenceofsirefertilitystatusonconceptusinducedtranscriptomicresponseofthebovineendometrium