College of Animal Science and Technology, Hebei Normal University of Science & Technology, Hebei Key Laboratory of Special Animal Germplasm Resources Mining and Innovation, Qinhuangdao, Hebei 066004, China.
College of Animal Science and Technology, Hebei Normal University of Science & Technology, Hebei Key Laboratory of Special Animal Germplasm Resources Mining and Innovation, Qinhuangdao, Hebei 066004, China.
Gene. 2025 Jan 20;934:149045. doi: 10.1016/j.gene.2024.149045. Epub 2024 Oct 24.
Melanocytes, which are mainly found in the epidermis, are responsible for the melanin of skin and hair, and thereby contribute to the appearance of skin and provide protection from damage by ultraviolet radiation. Our previous study revealed that the Wnt5a, one of the many genes that affect melanin production, might be involved in the coat color seasonal change of the Arctic fox by influencing skin melanogenesis. Although the role of Wnt5a in melanocyte lines and melanoma cells has been extensively studied, its role in primary epidermal melanocytes has not been explored. This study aimed to investigate the role and mechanism of the Wnt5a in influencing melanogenesis in Arctic fox primary epidermal melanocytes. We constructed the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) knockout plasmid targeting exons of the Wnt5a and transfected it into primary epidermal melanocytes. The results of the amplification knockout region assay, RT-qPCR assay, and western blot assay showed the success of Wnt5a knockout. RT-qPCR assay and melanin content assay showed that melanin production in melanocytes was significantly increased after Wnt5a knockout, and melanin-related key genes, such as microphthalmia-associated transcription factor, tyrosinase and tyrosinase-related protein 1, were significantly elevated. In addition, we also found that the expression of the β-catenin gene of the Wnt canonical pathway was significantly elevated after Wnt5a knockout. In conclusion, our results indicate that the Wnt5a plays a negative regulatory role in melanogenesis in primary epidermal melanocytes, and is presumably involved in antagonizing or inhibiting canonical Wnt signaling.
黑素细胞主要存在于表皮中，负责皮肤和毛发的黑色素生成，从而影响皮肤外观，并提供抵御紫外线辐射的保护。我们之前的研究表明，Wnt5a 是影响黑色素生成的众多基因之一，可能通过影响皮肤黑色素生成而参与北极狐的毛色季节性变化。虽然 Wnt5a 在黑素细胞系和黑色素瘤细胞中的作用已得到广泛研究，但它在原代表皮黑素细胞中的作用尚未得到探索。本研究旨在探讨 Wnt5a 影响北极狐原代表皮黑素细胞黑色素生成的作用和机制。我们构建了靶向 Wnt5a 外显子的 CRISPR 敲除质粒，并将其转染到原代表皮黑素细胞中。扩增敲除区域检测、RT-qPCR 检测和 Western blot 检测结果表明 Wnt5a 敲除成功。RT-qPCR 检测和黑色素含量检测结果表明，Wnt5a 敲除后黑素细胞中黑色素生成显著增加，微phthalmia 相关转录因子、酪氨酸酶和酪氨酸酶相关蛋白 1 等黑色素相关关键基因显著升高。此外，我们还发现 Wnt 经典途径的 β-catenin 基因的表达在 Wnt5a 敲除后显著升高。总之，我们的结果表明 Wnt5a 在原代表皮黑素细胞的黑色素生成中起负调控作用，可能参与拮抗或抑制经典 Wnt 信号。
