In ovo injection of AZD6244 suppresses feather follicle development by the inhibition of ERK and Wnt/β-catenin pathways in goose embryos (Anser cygnoides)

Abstract

ABSTRACT 1. Feathers are an important product from poultry, and the state of feather growth and development plays an important role in their economic value. 2. In total, 120 eggs were selected for immunoblotting and immunolocalisation experiments of ERK and β-catenin proteins in different developmental stages of goose embryos. The ERK protein was highly expressed in the early stage of goose embryo development, while β-catenin protein was highly expressed in the middle stage of embryo development. 3. The 120 eggs were divided into four treatment groups, including an uninjected group (BLANK), a group injected with 100 µl of cosolvent (CK), a group injected with 100 µl of AZD6244 containing cosolvent in a dose of 5 mg/kg AZD6244 containing cosolvent (AZD5) and a group injected with 100 µl of AZD6244 containing cosolvent in a dose of 15 mg/kg AZD6244 containing cosolvent (AZD15). The eggs were injected on the ninth day of embryonic development (E9). Samples were collected at E21.5 to observe feather width, feather follicle diameter, ERK and Wnt/β-catenin pathway protein expression. 4. The AZD5 and AZD15 doses were within the embryonic safety range compared to the BLANK and CK groups and had no significant effect on the survival rate and weight at the inflection point, but significantly reduced the feather width and feather follicle diameter (p < 0.05). The AZD6244 treatment inhibited ERK protein phosphorylation levels and blocked the Wnt/β-catenin pathway, which in turn significantly down-regulated the expression levels of FZD4, β-catenin, TCF4 and LEF1 (p < 0.05), with an inhibitory effect in the AZD15 group being more significant. The immunohistochemical results of β-catenin and p-ERK were consistent with Western blot results. 5. The small molecule inhibitor AZD6244 regulated the growth and development of feather follicles in goose embryos by the ERK and Wnt/β-catenin pathways.