Dynamic epigenetic and transcriptomic reprogramming during embryonic skin development in goose (Anser anser domesticus).

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Feather follicles are specialized skin appendages that are essential for thermoregulation, protection, and down production in birds, forming through complex genetic and epigenetic interactions during embryogenesis. In this study, we examined skin and follicle development in Hungarian white goose embryos, focusing on dynamic epigenetic-transcriptomic changes. Histology showed smooth epidermis at E10, feather buds at E13, and columnar follicles with medullary tissue and secondary follicles at E18. Transcriptomics revealed 1327 and 1847 DEGs enriched in epidermal development, differentiation, and adhesion. Primordial initiation at E10-E13 featured Wnt, TGF-β, and melanogenesis, whereas follicle formation at E13-E18 involved lipid metabolism and VEGF signaling. Keratinization genes were continuously upregulated, and Wnt, Shh, and muscle pathways were activated late. Key regulators included LEF1, MSX2, and FOXN1. ATAC-seq showed dynamic chromatin accessibility, stage-specific promoter openness, and motifs for YY1, KLF5, and KLF4. Differentially accessible regions enriched genes in Wnt and TGF-β signaling, cell adhesion, and mitophagy, shifting from proliferation and basic metabolism at E10-E13 to differentiation, lipid metabolism, and homeostasis at E13-E18. Integrated analyses linked fatty acid metabolism, MAPK, and FoxO signaling to differentiation, while downregulation of redox and migration pathways preserved homeostasis. Some fatty acid metabolism and cell polarity genes increased expression despite reduced accessibility at E13-E18, indicating epigenetic pre-programming and post-transcriptional interplay. This work delineates coordinated epigenetic-transcriptional regulation of goose embryonic skin and feather follicle morphogenesis, offering insights for avian and vertebrate skin appendage studies.

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1. The Forkhead box O3 (FOXO3) transcription factor is a crucial regulator in controlling cell metabolism, proliferation, apoptosis, migration and response to oxidative stress. However, FOXO3 has not previously been studied much in the embryonic skin follicles of geese. 2. This study used Zhedong white geese (Anser cygnoides), Jilin white geese (Anser cygnoides) and Hungarian white geese (Anser anser). The feather follicle structure in the dorsal skin during embryonic stages was examined with haematoxylin and eosin (HE) and Pollak staining. The FOXO3 protein content in the embryonic dorsal skin from feather follicles was detected using western blotting and quantitative real-time PCR. 3. The mRNA expression level of FOXO3 in the dorsal skin of Jilin white geese was highly expressed on embryonic day 23 (E23; P < 0.01), while mRNA expression of FOXO3 was highly expressed in the feather follicle of Hungarian white geese at E28 (P < 0.01). The expression of FOXO3 protein mainly concentrated in the early embryonic phase among these goose breeds (P < 0.05). This suggested that FOXO3 plays a crucial role in the development and growth of embryonic dorsal skin of feather follicles. The location of the FOXO3 protein was determined using the IHC technique, which further verified the effect of FOXO3 in the dorsal skin for feather follicles during embryogenesis. 4. The study demonstrated the differential expression and localisation of the FOXO3 gene among different goose species. It was speculated that the gene could potentially improve goose feather follicle development and feather-related traits and provide a basis for further understanding of FOXO3 function in the dorsal tissue of goose embryos.

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