Subfertility of Offspring and Transgenerational Transmission of Splenic Transcriptome Induced by Lipopolysaccharide in Hens
Authors: Liu Lei , Wang Di , Li Xingzheng , Adetula Adeyinka Abiola , Khan Adnan , Xu Guiyun , Yang Ning , Yu Ying 
Published: May 2021

Abstract
Background: Transgenerational phenotypes are linked to genetic background, environmental factors, and diseases. Maternal stimulation is one of the suspected contributing factors to transgenerational phenotypes, and recent evidence from human studies suggests that maternal immune stimulation affects the phenotype and gene expression of the offspring, but few similar studies were reported in poultry. Here, we used laying hens as a model organism to investigate the effects of maternal stimulation on hens' immunity and reproductive performance, as well as the regulatory role of the splenic transcriptome in hens induced by lipopolysaccharide (LPS).Methods: To investigate the effect of maternal stimulation on egg-laying rate of hens and their offspring, laying hens were first intravenously injected with LPS. RNA-seq from the spleens of F0, F1, and F2 generations of hens was used to identify differentially expressed genes (DEGs) between the LPS group and controls. We investigated the effect of LPS maternal stimulation on the transcriptome of laying hens and its multi-generational transmission effect by analyzing shared genes, enrichment pathways, and protein-protein interaction networks across generations. We concentrated on immune and reproduction-related pathways and genes that existed across generations. Subsequently, we performed a correlation analysis between the DEGs associated with the multi-generational transmission effect and egg-laying rate of hens.Results: We found that the LPS maternal stimulation could reduce the egg-laying rate of hens and their offspring, especially during the early and late laying stages. The transcriptome study of the spleen in F0, F1 and F2 generations showed that the maternal stimulation of the LPS affects the patterns of gene expression in laying hens, and this change has the effect of transgenerational transmission. Further analysis of DEGs and their enrichment pathways found that the LPS maternal stimulation mainly affects the reproduction and immunity of laying hens and their offspring. The DEGs such as AVD, HPS5 CATHL2, S100A12, EXFABP, RSFR, LY86, PKD4, XCL1, FOS, TREM2 and MST1 may play an essential role in the regulation of the immunity and egg-laying rate of the hens. Furthermore, the MMR1L3, C3, F13A1, LY86 and GDPD2 genes with transgenerational transmission effects are highly correlated with the egg-laying rate. The DEGs mentioned above have an important reference value for research on the multi-generational transmission of maternal immune stimulation.Conclusions: In the current study, we discovered that maternal stimulation could reduce the immunity of laying hens and their offspring, resulting in a decrease in egg-laying rate. These effects could be regulated by the transcriptome's transgenerational transmission. Overall, our study is an important reference for future research into the multi-generational transmission of maternal stimulation, and the selected marker genes are of great significance to the breeding of laying hens.