High-fat Diet Alters Male Seminal Plasma Composition to Impair Female Immune Adaptation for Pregnancy in Mice

Abstract

Paternal experiences and exposures prior to conception can influence fetal development and offspring phenotype. The composition of seminal plasma contributes to paternal programming effects, through modulating the female reproductive tract immune response after mating. To investigate whether paternal obesity affects seminal plasma immune-regulatory activity, C57Bl/6 male mice were fed an obesogenic high fat diet (HFD) or control diet (CD) for 14 weeks. While HFD consumption caused only minor changes to parameters of sperm quality, the volume of seminal vesicle fluid secretions was increased by 65%, and the concentrations and total content of immune-regulatory TGFB isoforms were decreased by 75-80% and 43-55% respectively. Mating with BALB/c females revealed differences in the strength and properties of the post-mating immune response elicited. Transcriptional analysis showed >300 inflammatory genes were similarly regulated in the uterine endometrium by mating independently of paternal diet, but 13 were dysregulated by HFD-fed compared to CD-fed males. Seminal vesicle fluid factors reduced in HFD-fed males, including TGFB1, IL10, and TNF, were amongst the predicted upstream regulators of differentially regulated genes. Additionally, the T cell response induced by mating with CD-fed males was blunted after mating with HFD-fed males, with 27% fewer CD4 + T cells, 26% fewer FOXP3 +CD4 + regulatory T cells (Treg) cells, and 19% fewer CTLA4 + Treg cells, particularly within the NRP1 + thymic Treg cell population. These findings demonstrate that an obesogenic high fat diet alters the composition of seminal vesicle fluid and impairs seminal plasma capacity to elicit a favorable pro-tolerogenic immune response in females at conception.

Elucidation of the protein composition of mouse seminal vesicle fluid

“…health [4][5][6][7]. Notably, SVF composition displays considerable plasticity in response to stressors as diverse as diet [8,9], social status [10,11], and cancer [12]. This sensitivity appears to provide a mechanism by which paternal environmental exposures and health status can act to influence female reproductive receptivity and offspring phenotype [6,13].…”

“…In this context, a variety of approaches have been utilized, including both gel-based [12,14] and gel-free [10,15] proteomics, but only a small number of SVF proteins have been identified to date [10,12,15]. Although this suggests low protein diversity in SVF, targeted analysis of specific proteins not detected in proteomics analyses, notably cytokines [6,9,16,17] and other immune-regulatory and antimicrobial proteins [13,18], indicates there is more complexity than is currently curated. Accordingly, the existing limits on proteomic depth likely reflect the fact that SVF has a unique, highly biased protein signature, characterized by a small subset of dominant proteins that mask detection of less abundant proteins.…”

“…To address this, we collected SVF from male outbred Swiss mice ( As previous proteomics studies identified intracellular proteins in SVF [9,11,21], microscopic analysis was used to exclude the presence of contaminating cells introduced during isolation (n = 3, Figure S1A).…”

“…This necessitated a vigorous solubilization step to allow comprehensive analysis of SVF composition. One approach that we and others have utilized is to solubilize SVF in the strong chaotropic agent guanidine hydrochloride [9,11,21,22]. However, guanidine hydrochloride is not commonly used in proteomics as it hampers trypsin activity, thereby limiting the efficiency of peptide digestion and consequently, the number of proteins identified [23].…”

“…Notably, lowly abundant proteins known to be carried by SVF, including cytokines, were not detected in our analysis. This is likely explained by the fact that cytokines are generally challenging to detect using MS due to their small size and low abundance, necessitating the need for alternative approaches to detect these molecules [9,17,41]. S3 We next used IPA to explore the functions of seminal vesicle secreted proteins.…”