In Ovo Injection of CHIR-99021 Promotes Feather Follicle Development via Modulating the Wnt Signaling Pathway and Transcriptome in Goose Embryos (Anser cygnoides).

Feather performs important physiological functions in birds, and it is also one of the economic productions in goose farming. Understanding and modulating feather follicle development during embryogenesis are essential for bird biology and the poultry industry. CHIR-99021 is a potent Wnt/ß-catenin signaling pathway activator associated with feather follicle development. In this study, goose embryos (Anser cygnoides) received an in ovo injection of CHIR-9902, which was conducted at the beginning of feather follicle development (E9). The results showed that feather growth and feather follicle development were promoted. The Wnt signaling pathway was activated by the inhibition of GSK-3ß. Transcriptomic analyses showed that the transcription changes were related to translation, metabolism, energy transport, and stress in dorsal tissue of embryos that received CHIR-99021, which might be to adapt and coordinate the promoting effects of CHIR-99021 on feather follicle development. This study suggests that in ovo injection of CHIR-99021 is a potential strategy to improve feather follicle development and feather-related traits for goose farming and provides profiling of the Wnt signaling pathway and transcriptome in dorsal tissue of goose embryos for further understanding of feather follicle development.

In ovo injection of CHIR-99021 promotes feather follicles development via activating Wnt/ß-catenin signaling pathway during chick embryonic period.

The Wingless-types/beta-catenin (Wnt/ß-catenin) signaling pathway plays an important role in embryonic development and affects the physiological development processes of feather follicles. To investigate the role of Wnt/ß-catenin pathway in regulating feather follicles morphogenesis, in ovo injection of CHIR-99021, an activator of the Wnt/ß-catenin signaling pathway, was conducted in chick embryo model. Initially, a total of 40 embryos were used to assess feather follicles morphogenesis and the expression of ß-catenin (E9-E17). The histological results showed that feather follicle morphogenesis was mainly completed from E9 to E17. ß-catenin was involved in the processing of the appearance of dermal cell condensation (E9) and the completion of the feather follicles morphogenesis (E17). Next, a total of 160 fertilized eggs were randomly divided into 8 groups for in ovo injection at E9, including a Normal Saline injected group (CON) and the 500, 1,000, 2,000, 5,000, 10,000, 50,000, and 100,000 ng CHIR-99021 groups. Dorsal skin tissue samples were collected at E17 for investigating feather follicles morphology and expressions of ß-catenin and lymphoid enhancerbinding factor-1 (LEF1) at gene and protein levels. The results showed that feather follicle diameter in the injected groups were significantly (P < 0.05) increased with limit dose-independence compared to the CON group. CHIR-99021 significantly (P < 0.05) influenced the mRNA expressions of catenin beta-1 (CTNNB1) and downstream target LEF1. In ovo injection of CHIR-99021 caused that ß-catenin and LEF1 were significantly (P < 0.05) increased followed the increased doses as determined by western blotting. The immunochemical results showed that ß-catenin was detected in the dermal papilla of feather follicles. Given these results, this study suggests to developmental biology that in ovo injection of CHIR-99021 promoted feather follicles morphogenesis and development via activating Wnt/ß-catenin signaling pathway and upregulating downstream target LEF1 during embryonic period in chick embryo model. Moreover, CHIR-99021 may be a strong candidate to promote the animal feather/hair industry, especially as a reference for bird feather production.

The role of CTNNB1 and LEF1 in feather follicles development of Anser cygnoides and Anser anser.

BACKGROUND: Wingless-types/beta-catenin (Wnt/ß-catenin) signaling pathway is one of the most extensively studied transcriptional cascades involved in various types of organogenesis including embryonic and postnatal development. Downy feather quantity is primarily affected by follicular development and gene regulations. OBJECTIVE: This research was aimed to investigate the role of catenin beta-1(CTNNB1) and lymphoid enhancerbinding factor-1 (LEF1) on feather follicles development at different developmental stages. METHODS: Fluorescence quantitative PCR, Western-blot and immunohistochemical methods were used in Anser cygnoides and Anser anser embryos (E12, E13 E18, and E28) and after birth gosling stages (G18, G48, G88) for gene expression analysis. RESULTS: CTNNB1 and LEF1 genes were expressed in Anser cygnoides and Anser anser at different embryonic and after-birth gosling developmental stages and the expression levels were significantly different in different stages (p < 0.05). The mRNA expression of CTNNB1 and LEF1 genes reached the highest level at D88 in Anser cygnoides, while the highest expression levels were at D18 and D88 in Anser anser, and the expression levels of CTNNB1 genes at D88 in all embryonic stages were significantly lower than after-birth stages. CTNNB1 and LEF1 protein expression were the highest at E12 and E28 for Anser cygnoides feather follicles development. While at a similar stage for Anser anser, the expression of CTNNB1 and LEF1 protein was the highest at D48 and D18. Protein expression at embryonic stages was in the epidermis (E) and the hair basal plate (P), the expression site for after-birth stages was in the dermal papilla (DP). CONCLUSION: Our study illustrated that CTNNB1 and LEF1 has an impact on Anser cygnoides and Anser anser feather follicles growth and development.