Proteomic markers of low and high fertility bovine spermatozoa separated by Percoll gradient

Abstract

In the context of artificial insemination, male fertility is defined as the ability to produce functional spermatozoa able to withstand cryopreservation. We hypothesized that interindividual variations in fertility depend on the proportion of the fully functional sperm population contained in the insemination dose. The objective of this study was to identify protein markers of the fully functional sperm subpopulation. Insemination doses from four high‐fertility (HF) and four low‐fertility (LF) bulls with comparable post‐thaw quality parameters were selected for proteomic analysis using iTRAQ technology. Thawed semen was centrifuged through a Percoll gradient to segregate the motile (high density [HD]) from the immotile (low density [LD]) sperm populations. Sperm proteins were extracted with sodium deoxycholate and four groups were compared: LD and HD spermatozoa from LF and HF bulls. A total of 498 unique proteins were identified and quantified. Comparison of HD spermatozoa from HF and LF bulls revealed that five proteins were significantly more abundant in the HF group (AK8, TPI1, TSPAN8, OAT, and DBIL5) whereas five proteins were more abundant in the LF group (RGS22, ATP5J, CLU, LOC616319, and CCT5). Comparison of LD spermatozoa from HF and LF bulls revealed that four proteins were significantly more abundant in the HF group (IL4I1, CYLC2, OAT, and ARMC3) whereas 15 proteins were significantly more abundant in the LF group (HADHA, HSP90AA1, DNASE1L3, SLC25A20, GPX5, TCP1, HIP1, CLU, G5E622, LOC616319, HSPA2, NUP155, DPY19L2, SPERT, and SERPINE2). DBIL5, TSPAN8, and TPI1 showed potential as putative markers of the fully functional sperm subpopulation.

Proteostasis regulated by testis-specific ribosomal protein RPL39L maintains mouse spermatogenesis

“…Disruption of spermatogenesis at any stage due to either genetic or environmental interventions may render male sterility and, in humans, accounts for 50% of infertility ( Krausz and Riera-Escamilla, 2018 ; Visser and Repping, 2010 ). Although it is conceivable that synthesis and quality of sperm proteome are relevant to sperm quality and male fertility, little is known about how dynamic proteostasis is achieved and maintained during spermatogenesis ( D'Amours et al., 2019 ; Netherton et al., 2018 ; Schafer et al., 1995 ).…”

Supplementation of cryoprotectant withPinus massonianabark extract improves human sperm vitality and fertility potential

“…Proteomics is an effective tool for sperm analysis and has been used widely to explore the differential proteins between populations with different fertility levels 43,44 or spermatozoa with different motility 45 . Recent proteomics studies of fresh spermatozoa and resuscitated spermatozoa after freezing have suggested that some sperm proteins are associated with sperm quality, and the loss of these proteins may be responsible for the decrease in fertility in cryopreserved spermatozoa 46‐48 .…”

ARMC Subfamily: Structures, Functions, Evolutions, Interactions, and Diseases

“…This finding suggests that the structure between poly-Ser and poly-Glu may play a special role in evolution. Moreover, it has been reported that the expression of ARMC3 is significantly increased in highly active sperm ( D’Amours et al, 2019 ), whereas the deletion of exon 11 of the ARMC3 gene may alter the reading frame, leading to premature termination of translation, which causes sperm defects and thus male infertility ( Pausch et al, 2016 ). Interestingly, ARMC3 exon 11 encodes amino acids 476–521 of ARMC3, which are located across the end of the last armadillo repeat sequence ( Figures 1 , 2 ).…”