A Novel Single-Nucleotide Polymorphism in WNT4 Promoter Affects Its Transcription and Response to FSH in Chicken Follicles

Abstract

The signaling pathway of the wingless-type mouse mammary tumor virus integration site (Wnt) plays an important role in ovarian and follicular development. In our previous study, WNT4 was shown to be involved in the selection and development of chicken follicles by upregulating the expression of follicle-stimulating hormone receptors (FSHR), stimulating the proliferation of follicular granulosa cells, and increasing the secretion of steroidal hormones. FSH also stimulates the expression of WNT4. To further explore the molecular mechanism by which FSH upregulates WNT4 and characterize the cis-elements regulating WNT4 transcription, in this study, we determined the critical regulatory regions affecting chicken WNT4 transcription. We then identified a single-nucleotide polymorphism (SNP) in this region, and finally analyzed the associations of the SNP with chicken production traits. The results showed that the 5′ regulatory region from −3354 to −2689 of WNT4 had the strongest activity and greatest response to FSH stimulation, and we identified one SNP site in this segment, −3015 (G > C), as affecting the binding of NFAT5 (nuclear factor of activated T cells 5) and respones to FSH stimulation. When G was replaced with C at this site, it eliminated the NFAT5 binding. The mRNA level of WNT4 in small yellow follicles of chickens with genotype GG was significantly higher than that of the other two genotypes. Moreover, this locus was found to be significantly associated with comb length in hens. Individuals with the genotype CC had longer combs. Collectively, these data suggested that SNP−3015 (G > C) is involved in the regulation of WNT4 gene expression by responding FSH and affecting the binding of NFAT5 and that it is associated with chicken comb length. The current results provide a reference for further revealing the response mechanism between WNT and FSH.

Untargeted Metabolomics Revealed Potential Biomarkers of Small Yellow Follicles of Chickens during Sexual Maturation

“…Ferroptosis is a newly discovered type of cell death that mainly depends on iron-mediated oxidative damage and subsequent cell membrane damage [ 56 ] and mainly manifests as glutathione (GSH) depletion and glutathione peroxidase 4 (GPX4) inactivation [ 57 , 58 ]. Previous research has shown that the TGF signaling pathway [ 59 ], Wnt signaling pathway [ 60 ], and cAMP pathway [ 61 ] are involved in the regulation of the follicle selection process in domestic chickens. Some studies have focused on ferroptosis induced by oxidative stress in the aquaculture industry and found that heat shock protein (HSP) , activating transcription factor 4 (ATF4) , solute carrier family 7 member 11 (SLC7A11), and GPX4 were associated with ferroptosis and the mitochondrial pathway [ 62 , 63 , 64 ].…”

MFN2 regulates progesterone biosynthesis and proliferation of granulosa cells during follicle selection in hens

“…In sexually mature hens, one of the SYFs are selected to develop into hierarchical follicles daily or near‐daily, which initiates hierarchical development under the regulation of FSH (Wang et al, 2022; Zhong et al, 2022). Generally, there are 5–6 hierarchical follicles of increased sizes, i.e., F6/F5 to F1, and 8–13 SYFs in commercial hens (Álvarez & Hocking, 2012; Wang et al, 2021).…”

“…Studies on mammals demonstrated that MFN2 sustained the proliferation and differentiation of the granulosa cell by regulating its ATP production and metabolism (Feng et al, 2022;Liu et al, 2014). Herein, this study aimed to investigate whether Zhong et al, 2022). Generally, there are 5-6 hierarchical follicles of increased sizes, i.e., F6/F5 to F1, and 8-13 SYFs in commercial hens (Álvarez & Hocking, 2012;Wang et al, 2021).…”