Characterization of the porcine seminal plasma proteome comparing ejaculate portions

Abstract

Full identification of SP-proteins remains challenging, particularly in some livestock species such as porcine. This experimental study aims to provide an extensive proteomic analysis of boar SP and to generate a public accessible database of boar SP-proteome. A SP-pool from 33 entire ejaculates from 11 boars (3 ejaculates per boar) was analyzed to characterize the boar SP-proteome. Moreover, 20 ejaculates collected in fractions (P1: first 10 mL of sperm rich ejaculate fraction (SRF), P2: rest of SRF and P3: post-SRF) from 5 boars (4 ejaculates per boar) were analyzed to evaluate differentially expressed SP-proteins among portions. SP-samples were subjected to a combination of SEC, 1-D SDS PAGE and NanoLC-ESI-MS/MS followed by functional bioinformatics analysis.The identified proteins were quantified from normalized LFQ intensity data. A total of 33,557 spectra corresponding to 8,189 peptides and 536 SP-proteins were identified with ≥ 95% Confidence (Unused Score > 1.3) and a false discovery rate (FDR) ≤ 1%. Of the 536 SP-proteins, 409 were identified in Sus Scrofa taxonomy and 374 of them were Biological Significance: This proteomic study provides the major characterization of the boar SP-proteome with more than 250 proteins first reported. The boar SP-proteome is described so that a spectral library can be built for relative 'label free' protein quantitation with SWATH approach. This proteomic profiling allows the creation of a publicly accessible database of the boar SP-proteome, as a first step for further understanding the role of SP-proteins in reproductive outcomes as well as for identification of biomarkers for sperm quality and fertility.

Seminal Influence of the Oviduct : Mating and/or semen components induce gene expression changes in the pre-ovulatory functional sperm reservoir in poultry and pigs

“…The studies in this thesis have shown that the composition of the seminal fluid in chicken with differences in fertility (RJF vs WL/AIL) varies in relation to protein type and contents, similarly to what is already known for pigs (Perez-Patiño et al, 2016). Comparative analyses of a battery of cytokines/chemokines in the SP/SF of boars and cocks also showed major differences.…”

“…The proteome of the boar seminal plasma has been, very recently, characterized in detail for all ejaculated fractions and further for their relation with the fertility of the sires (Perez-Patiño et al, 2016). In pig, a convenient animal model for studies on species with fractionated ejaculates as human, the SP has been found to be involved in the induction of environmental changes in the female genitalia, previously considered as a sperm-related role, regarding pro-and anti-inflammatory phenomena (Lovell & Getty, 1968, Rodriguez-Martinez et al, 1990, Rozeboom et al, 1999.…”

“…The SF/SP signaling proteins and peptides -i.e. avian β-defensins (Marzoni et al, 2013;Labas et al, 2015); or porcine spermadhesins (Rodríguez-Martínez et al, 2010;Perez-Patiño et al, 2016); cytokines/chemokines (pig: O´Leary et al, 2011;Jiwakanon and Dalin, 2012), and or sex hormones (progesterone, testosterone, dihydrotestosterone and estrogen) (chicken: Anderson and Navara, 2011;Oliveira et al, 2011;pig: Claus et al, 1983;Zdunczyk et al, 2011), are all considered to contribute to the orchestration of such local immune defence responses in the female genitalia.…”

“…These proteins are involved in a variety of effects on sperm protection including membrane stabilization, capacitation, and sperm-oviduct/oocyte interactions (Calvete et al 1997;Rodriguez-Martinez et al, 1998a;Töpfer-Petersen et al, 1998;Calvete et al, 2005;Caballero et al, 2006), as well as immune stimulation in vivo (Rodríguez-Martínez et al, 2010). In the boar, whose fractionated ejaculate sequence mimics that of human, its SP-proteome is rather well described (Rodríguez-Martínez et al, 2009;Rodriguez-Martinez et al, 2011;Patiño et al, 2016). Yet, the peptidome of cytokines and chemokines is less screened, with only few cytokines identified.…”

Markers for Sperm Freezability and Relevance of Transcriptome Studies in Semen Cryopreservation: A Review

“…It is worth noting that proteins that are involved in the energy metabolism of spermatozoa play key roles in glycolysis, the citric acid cycle, and oxidative phosphorylation, which are required to provide suicient energy for the sperm physiological functions [37]. The protein constellation of the SP and sperm cells, and their distinct subcompartments have been well documented in a numerous animal species, using a plethora of proteomicbased techniques [6,37,38]. While some speciic sperm protein markers facilitating good semen freezability have been identiied [10,24,39,40], their function depends on the presence of mRNA that can be translated into proteins in the spermatozoa.…”