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

Beatriz Fernandez-Fuertes,José María Sánchez,Sandra Bagés-Arnal,Marc Yeste,Pat Lonergan,Michael Mcdonald

doi:10.1038/s41598-019-51413-4

Beatriz Fernandez-Fuertes, José María Sánchez + Show 4 more

Open Access

https://doi.org/10.1038/s41598-019-51413-4

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Journal: Scientific Reports	Publication Date: Oct 21, 2019
Citations: 10	License type: open-access

Affiliation: University of Girona, University College Dublin

Abstract

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.

Highlights

During semen ejaculation in mammals, epididymal sperm are mixed with secretions of the accessory sex glands, collectively termed seminal plasma (SP)
A high variability was found in the relative mRNA abundance of interleukins-1A, -B, -8, and -6 (IL1A, IL1B, IL8, IL6), prostaglandin E synthase 2 (PTGES2), tumor necrosis factor A (TNFA), and leukemia inhibitory factor (LIF) between animals, but no effect of treatment was observed in comparison with the control
Endometrial explants from heifers collected at a commercial slaughterhouse (n = 3) were incubated for 6 h with Roswell Park Memorial Institute-1640 (RPMI) media alone, or SP which had previously been treated for 10 min at 60 °C, 45 °C or 39 °C with or without a commercial RNase inactivation agent. (D) Experimental design for Experiment 4

Summary

Introduction

During semen ejaculation in mammals, epididymal sperm are mixed with secretions of the accessory sex glands, collectively termed seminal plasma (SP). The specific objectives of the study were to determine: (i) gene expression changes in bovine endometrium due to exposure to sperm or SP (Experiment 1); (ii) whether SP from an intravaginal ejaculator (bull) or an intrauterine ejaculator (stallion) had a differential effect on endometrial RNA quality, and whether this effect was concentration-dependent (Experiment 2); (iii) whether an RNase inactivation reagent could block the detrimental effect on RNA quality induced by bull SP (Experiment 3); (iv) whether semen collection method alters the effect of SP on the endometrium (Experiment 4): and (v) whether tissues that physiologically (i.e., during natural mating) come in direct contact with bovine SP (i.e., vagina and cervix) exhibit more resilience than the endometrium (Experiment 5)

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