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Seminal plasma, and not sperm, induces time and concentration-dependent neutrophil extracellular trap release in donkeys

Digital.CSIC. Repositorio Institucional del CSIC
  • Mateo-Otero, Yentel
  • Zambrano, Fabiola
  • Catalán, Jaime
  • Sánchez, Raúl
  • Yeste, Marc
  • Miro, Jordi
  • Fernández-Fuertes, Beatriz
12 Pág., In several mammalian species, acute endometritis driven by the recruitment of polymorphonuclear cells (PMN) occurs in response to semen. These PMNs release DNA to form neutrophil extracellular traps (NETs) in cattle, horse and human, leading to sperm entrapment. While there is no evidence of this phenomenon occurring in donkeys, artificial insemination (AI) with frozen-thawed semen, which results in very poor pregnancy rates, leads to a large PMN recruitment to the uterus., The Ministry of Science and Innovation, Spain (grants: RYC-2014-15581 and AGL2017-88329-R), the Regional
Government of Catalonia (grants: 2019-SGR-1229 and 2020-FI-B-00412) and the European Union's Horizon 2020 Research and Innovation Program (grant: 792212)., Peer reviewed




Sperm exposure to accessory gland secretions alters the transcriptomic response of the endometrium in cattle

Digital.CSIC. Repositorio Institucional del CSIC
  • Sánchez, José María
  • Rabaglino, M. B.
  • Bagés Arnal, Sandra
  • McDonald, Michael
  • Behura, Susanta K.
  • Spencer, T. E.
  • Lonergan, P.
  • Fernández-Fuertes, Beatriz
9 Pàg., In cattle, mating to intact, but not vasectomised, bulls has been shown to modify the endometrial transcriptome, suggesting an important role of sperm in the modulation of the uterine environment in this species. However, it is not clear whether these changes are driven by intrinsic sperm factors, or by factors of accessory gland (AG) origin that bind to sperm at ejaculation. Therefore, the aim of the present study was to determine whether ejaculated sperm, which are suspended in the secretions of the AGs, elicit a different endometrial transcriptomic response than epididymal sperm, which have never been exposed to AG factors. To this end, bovine endometrial explants collected from heifers in oestrus were (co-)incubated for 6 h alone (control), or with epididymal sperm or ejaculated sperm, following which transcriptomic changes in the endometrium were evaluated. Epididymal sperm elicited a more dramatic endometrial response than ejaculated sperm, in terms of the number of differentially expressed genes (DEGs). Indeed, RNA-sequencing data analysis revealed 1912 DEGs in endometrial explants exposed to epididymal sperm compared with control explants, whereas 115 DEGs were detected between endometrial explants exposed to ejaculated sperm in comparison to control explants. The top pathways associated with genes upregulated by epididymal sperm included T cell regulation and TNF, NF-KB and IL17 signalling. Interestingly, ejaculated sperm induced downregulation of genes associated with T cell immunity and Th17 differentiation, and upregulation of genes involved in NF-KB signalling, in comparison to epididymal sperm. These data indicate that factors of AG origin modulate the interaction between sperm and the endometrium in cattle., The authors wish to thank the National Cattle Breeding Centre for donating the fresh bull ejaculates used in this study, and Dr. John A. Browne (UCD) for his technical assistance. This project has received funding from the European Union's Horizon 2020 research and innovation program (Grant No. 792212); Science Foundation Ireland (Grant No. 16/IA/4474); and the Spanish Ministry of Science and Innovation (Grant No. IJC2019-039034-I)., Peer reviewed




Species-specific and collection method-dependent differences in endometrial susceptibility to seminal plasma-induced RNA degradation

DUGiDocs – Universitat de Girona
  • Fernández Fuertes, Beatriz
  • Sánchez, José María
  • Bagés-Arnal, Sandra
  • McDonald, Michael
  • Yeste Oliveras, Marc
  • Lonergan, Pat
This study aimed to determine the effect of bull seminal plasma (SP) and sperm on endometrial function. Bovine endometrial explants were incubated with: ejaculated sperm with or without SP, epididymal sperm, or SP alone. Neither ejaculated nor epididymal sperm induced differential expression of IL1A, IL1B, IL6, IL8, PTGES2, TNFA, and LIF. Interestingly, SP had a detrimental effect on endometrial RNA integrity. Addition of an RNase inactivation reagent to SP blocked this effect, evidencing a role for a SP-RNase. Because bulls deposit the ejaculate in the vagina, we hypothesized that the bovine endometrium is more sensitive to SP-RNase than vaginal and cervical tissues (which come into contact with SP during mating), or to endometrium from intrauterine ejaculators (such as the horse). In addition, due to differences in SP-RNase abundance depending on SP collection method (i.e., with an artificial vagina, AV, or by electroejaculation, EE), this effect was also tested. Bull SP, collected by AV, degrades RNA of mare endometrium, and bovine vagina, cervix and endometrium. However, stallion SP or bull SP collected by EE did not elicit this effect. Thus, results do not support a role for SP in modulating endometrial function to establish pregnancy in cattle
Proyecto: EC/H2020/792212




Effect of exposure to seminal plasma through natural mating in cattle on conceptus length and gene expression

DUGiDocs – Universitat de Girona
  • Mateo Otero, Yentel
  • Sánchez, José María
  • Recuero, Sandra
  • Bagés-Arnal, Sandra
  • McDonald, Michael
  • Kenny, David A.
  • Yeste Oliveras, Marc
  • Lonergan, Pat
  • Fernández Fuertes, Beatriz
A growing body of evidence suggests that paternal factors have an impact on offspring development. These studies have been mainly carried out in mice, where seminal plasma (SP) has been shown to regulate endometrial gene expression and impact embryo development and subsequent offspring health. In cattle, infusion of SP into the uterus also induces changes in endometrial gene expression, however, evidence for an effect of SP on early embryo development is lacking. In addition, during natural mating, the bull ejaculates in the vagina; hence, it is not clear whether any SP reaches the uterus in this species. Thus, the aim of the present study was to determine whether SP exposure leads to improved early embryo survival and developmental rates in cattle. To this end, Day 7 in vitro produced blastocysts were transferred to heifers (12-15 per heifer) previously mated to vasectomized bulls (n = 13 heifers) or left unmated (n = 12 heifers; control). At Day 14, heifers were slaughtered, and conceptuses were recovered to assess size, morphology and expression of candidate genes involved in different developmental pathways. Additionally, CL volume at Day 7, and weight and volume of CL at Day 14 were recorded. No effect of SP on CL volume and weight not on conceptus recovery rate was observed. However, filamentous conceptuses recovered from SP-exposed heifers were longer in comparison to the control group and differed in expression of CALM1, CITED1, DLD, HNRNPDL, PTGS2, and TGFB3. In conclusion, data indicate that female exposure to SP during natural mating can affect conceptus development in cattle. This is probably achieved through modulation of the female reproductive environment at the time of mating, This work was supported by the European Union’s Horizon
2020 Research and Innovation Program (Grant Agreement
No. 792212) and Science Foundation Ireland (Grant
No. 16/IA/4474)
Proyecto: EC/H2020/792212




Mating to intact, but not vasectomized, males elicits changes in the endometrial transcriptome: insights from the bovine model

DUGiDocs – Universitat de Girona
  • Recuero, Sandra
  • Sánchez, José María
  • Mateo Otero, Yentel
  • Bagés-Arnal, Sandra
  • McDonald, Michael
  • Behura, Susanta K.
  • Spencer, Thomas E.
  • Kenny, David A.
  • Yeste Oliveras, Marc
  • Lonergan, Pat
  • Fernández Fuertes, Beatriz
Les dades d’expressió gènica estan disponibles públicament al repositori digital Dryad https://doi.org/10.5061/dryad.s7h44j14r i al repositori institucional DUGiDocs http://hdl.handle.net/10256/18379, An appropriate female reproductive environment is essential for pregnancy success. In several species, including mice, pigs and horses, seminal plasma (SP) components have been shown to modulate this environment, leading to increased embryo viability and implantation. Due to the characteristics of mating in the aforementioned species, SP comes into direct contact with the uterus. However, it is questionable whether any SP reaches the uterus in species that ejaculate inside the vagina, such as humans and cattle. Hence, we hypothesized that sperm, perhaps acting as a vehicle for SP factors, play a more important role in the modulation of the maternal uterine environment in these species. In addition, changes elicited by SP and/or sperm may originate in the vagina and propagate to more distal regions of the female reproductive tract. To test these hypotheses, a bovine model in which heifers were mated to intact or vasectomized bulls or were left unmated was used. RNA-sequencing of endometrial samples collected 24 h after mating with a vasectomized bull did not reveal any differentially expressed genes (DEGs) in comparison with control samples. However, the endometrium of heifers mated with intact bulls exhibited 24 DEGs when compared to heifers mated with vasectomized bulls, and 22 DEGs when compared to unmated control heifers. The expression of a set of cytokines (IL6, IL1A, IL8, and TNFA) and candidate genes identified in the endometrial RNA-sequencing (PLA2G10, CX3CL1, C4BPA, PRSS2, BLA-DQB, and CEBPD) were assessed by RT-qPCR in the vagina and oviductal ampulla. No differences in expression of these genes were observed between treatments in any region. However, mating to both intact and vasectomized bulls induced an increase in IL1A and TNFA expression in the vagina compared to the oviduct. These data indicate that sperm, but not secretions from the accessory glands alone, induce modest changes in endometrial gene expression after natural mating in cattle. However, it is not clear whether this effect is triggered by inherent sperm proteins or SP proteins bound to sperm surface at the time of ejaculation, This work was funded by the EU Horizon 2020 Marie Skłodowska-Curie (No. 792212) and Science Foundation Ireland (Grant No. 16/IA/4474)
Proyecto: EC/H2020/792212




RNA-sequencing endometrium intact, vasectomized, and control heifers [dades de recerca]

DUGiDocs – Universitat de Girona
  • Recuero, Sandra
  • Sánchez, José María
  • Mateo Otero, Yentel
  • Bagés-Arnal, Sandra
  • McDonald, Michael
  • Behura, Susanta K.
  • Spencer, Thomas E.
  • Kenny, David A.
  • Yeste Oliveras, Marc
  • Lonergan, Pat
  • Fernández Fuertes, Beatriz
Dades primàries associades a l'article publicat: Recuero, S., Sánchez, J.M., Mateo Otero, Y., Bagés-Arnal, S., McDonald, M., Behura, S.K., Spencer, T.E., Kenny, D.A., Yeste Oliveras, M., Lonergan, P., Fernández Fuertes, B. Mating to intact, but not vasectomized, males elicits changes in the endometrial transcriptome: insights from the bovine model. Frontiers in Cell And Developmental Biology, 2020, vol.8, art. núm. 547. Disponible a https://doi.org/10.3389/fcell.2020.00547, File with genetic expression data of endometrial samples from heifers mated with intact bulls, vasectomized bulls and unmated control heifers. Endometrial samples were collected 24h.after mating
Proyecto: EC/H2020/792212