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ABSTRACT: Spermatogenesis is a crucial indicator of geese reproduction performance and production. The testis is the main organ responsible for sperm production, and the egg-laying cycle in geese is a complex physiological process that demands precise orchestration of hormonal cues and cellular events within the testes, however, the seasonal changes in the transcriptomic and proteomic profiles of goose testicles remain unclear. To explore various aspects of the mechanisms of the seasonal cyclicity of testicles in different goose breeds, in this study, we used an integrative transcriptomic and proteomic approach to screen the key genes and proteins in the testes of 2 goose males, the Hungarian white goose and the Wanxi white goose, at 3 different periods of the laying cycle: beginning of laying cycle (BLC), peak of laying cycle (PLC), and end of laying cycle (ELC). The results showed that a total of 9,273 differentially expressed genes and 4,543 differentially expressed proteins were identified in the geese testicles among the comparison groups. The Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis suggested that the DEGs, in the comparison groups, were mainly enrichment in metabolic pathways, neuroactive ligand-receptor interaction, cyctokine-cyctokine receptor interaction, calcium signaling pathway, apelin signaling pathway, ether lipid metabolism, cysteine, and methionine metabolism. While the DEPs, in the 3 comparison groups, were mainly involved in the ribosome, metabolic pathways, carbon metabolism, proteasome, endocytosis, lysosome, regulation of actin cytoskeleton, oxidative phosphorylation, nucleocytoplasmic transport, and tight junction. The protein-protein interaction network analysis (PPI) indicated that selected DEPs, such as CHD1L, RAB18, FANCM, TAF5, TSC1/2, PHLDB2, DNAJA2, NCOA5, DEPTOR, TJP1, and RAPGEF2, were highly associated with male reproductive regulation. Further, the expression trends of 4 identified DEGs were validated by qRT-PCR. In conclusion, this work offers a new perspective on comprehending the molecular mechanisms and pathways involved in the seasonal cyclicity of testicles in the Hungarian white goose and the Wanxi white goose, as well as contributing to improving goose reproductive performance.

ABSTRACT: The Hungarian White Goose (Anser anser domesticus) is an excellent European goose breed, with high feather and meat production. Despite its importance in the poultry industry, no available genome assembly information has been published. This study aimed to present Chromosome-level and functional genome sequencing of the Hungarian White Goose. The results showed that the genome assembly has a total length of 1115.82 Mb, 39 pairs of chromosomes, 92.98% of the BUSCO index, and contig N50 and scaffold N50 were up to 2.32 Mb and 60.69 Mb, respectively. Annotation of the genome assembly revealed 19550 genes, 286 miRNAs, etc. We identified 235 expanded and 1,167 contracted gene families in this breed compared with the other 16 species. We performed a positive selection analysis between this breed and four species of Anatidae to uncover the genetic information underlying feather follicle development. Further, we detected the function of miR-199-x, miR-143-y, and miR-23-z on goose embryonic skin fibroblast. In summary, we have successfully generated a highly complete genome sequence of the Hungarian white goose, which will provide a great resource to improve our understanding of gene functions and enhance the studies on feather follicle development at the genomic level.

ABSTRACT: Goose down feather has become one of the most important economical products in the goose breeding industry and it provides several essential physiological roles in birds. Therefore, understanding and regulating the development of skin and feather follicles during embryogenesis is critical for avian biology and the poultry industry. MicroRNAs are known to play an important role in controlling gene expression during skin and feather follicle development. In this study, bioinformatics analysis was conducted to select miR-140-y as a potential miRNA involved in skin and feather follicle development and to predict TCF4 as its target gene. This gene was expressed at significant levels during embryonic feather follicle development, as identified by qPCR and Western blot. The targeting relationship was confirmed by a dual-luciferase assay in 293T cells. Then, the miR-140-y/TCF4 function in dermal fibroblast cells was explored. The results showed that miR-140-y could suppress the proliferation of goose embryonic dermal fibroblast cells (GEDFs) by suppressing the activity of some Wingless-types (Wnt) pathway related genes and proliferation marker genes, while miR-140-y inhibition led to the opposite effect. Similarly, the inhibition of the TCF4 gene results in blocking the proliferation of GEDFs by reducing the activity of some Wnt pathway-related genes. Finally, the co-transfection of miR-140-y inhibitor and siRNA-TCF4 results in a rescue of the TCF4 function and an increase of the Wnt signaling pathway and GEDFs proliferation. In conclusion, these results demonstrated that the miR-140-y-TCF4 axis influences the activity of the Wnt signaling pathway and works as a dynamic regulator during skin and feather follicle development.

ABSTRACT: Feather is an important economic trait of poultry, and growth and development state of feathers plays an important role in the economic value of poultry. Dermal fibroblasts are required for structural integrity of the skin and for feather follicle development. How FOXO3 affects feather follicle development as skin tissues change during goose embryo (Anser cygnoides) development and growth is not well understood. Here, we demonstrate that in vitro culture of single feathers and skin tissue results in changes in feather morphological structure by adding drugs to the culture medium that affect FOXO3 expression. We used feather follicles to show that during growth, the root location of feathers, the dermis layer, affects cell proliferation and apoptosis and regulates the expression of major genes in the Wingless-types/beta-catenin (Wnt/β-catenin) signaling pathway through the activity of FOXO3 in dermal fibroblasts. Feathers and dorsal skin tissues develop the correct structure, but feather length and width and feather follicle diameter change significantly (p < 0.05) without significant changes in feather follicle density (p > 0.05). Transfected dermal fibroblasts also showed that FOXO3 affected the formation and development of feather follicles in the embryonic stage by regulating the Wnt/β-catenin signaling pathway. Therefore, this study reveals the critical role of dermal fibroblast-FOXO3-induced Wnt/β-catenin signaling in promoting the formation and development of embryonic feather follicles.