Abstract
The molecular basis of male fertility remains unclear, especially in chickens, where decades of genetic selection increased male fertility variability as a side effect. As transcription and translation are highly limited in sperm, proteins are key molecules defining their functionality, making proteomic approaches one of the most adequate methods to investigate sperm capacity. In this context, it is interesting to combine complementary proteomic approaches to maximize the identification of proteins related to sperm-fertilizing ability. In the present study, we aimed at identifying proteins related to fertility in meat-type roosters, showing fertility variability. Fertile roosters (fertility rates higher than 70% after artificial insemination) differed from subfertile roosters (fertility rates lower than 40%) in their sperm mass motility. Fertile and subfertile sperm protein contents were compared using two complementary label-free quantitative proteomic methods: Intact Cell MALDI-TOF-Mass Spectrometry and GeLC-MS/MS. Combining the two strategies, 57 proteins were identified as differentially abundant. Most of them were described for the first time as differentially abundant according to fertility in this species. These proteins were involved in various molecular pathways including flagellum integrity and movement, mitochondrial functions, sperm maturation, and storage in female tract as well as oocyte–sperm interaction. Collectively, our data improved our understanding of chicken sperm biology by revealing new actors involved in the complexity of male fertility that depends on multiple cell functions to reach optimal rates. This explains the inability of reductionist in vitro fertility testing in predicting male fertility and suggests that the use of a combination of markers is a promising approach.
Highlights
Male fertility depends on the optimal functioning of all processes needed for sperm to fertilize the oocyte
7 presented a fold-change ratio (FC) ≤ 0.66 or ≥ 1.5 (Figure 2C). The annotation of these peaks was performed by computational confrontation against a homemade database of chicken sperm proteoforms and peptidoforms identified by Top-Down mass spectrometry (MS) (Soler et al, 2016a)
The molecular basis of male fertility remains unclear, especially in chickens, where decades of genetic selection induced an important increase in male fertility variability as a side effect (Soler et al, 2016a)
Summary
Male fertility depends on the optimal functioning of all processes needed for sperm to fertilize the oocyte. Proteomic analysis of sperm cells provides two types of knowledge: on the one hand, global proteome analysis informs on the molecular pathways essential for sperm biology, and on the other hand, it helps identify potential male fertility biomarkers. These biomarkers are largely needed to predict the individual fertility capacity in human medicine (Jodar et al, 2017), animal reproduction (Druart et al, 2019), and genetic conservation programs of endangered breeds and species (Soler et al, 2020). The Intact Cell MALDI-TOF Mass Spectrometry (ICM-MS) approach is a high-throughput methodology allowing the characterization of endogenous peptidoforms and proteoforms (
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