FOXL2 regulates the expression of the <i>Col4a1</i> collagen gene in chicken granulosa cells

Du, Hongting; Guo, Yan; Wu, Xiaohui; Gong, Yanzhang

doi:10.1002/mrd.23554

Cited by 10 publications

(6 citation statements)

References 21 publications

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“…In cattle, COL4A1 is a gene that stays in relatively low abundance throughout the follicle stages, with a decrease in P and an increase only observed in atretic follicles (28). COL4A1 is directly regulated by forkhead box L2 (FOXL2), a transcription factor speci cally expressed in granulosa cells (45). High levels of FOXL2 decrease COL4A1 (45), which is required for the proliferation of GC, allowing follicle remodeling, and should remain high during folliculogenesis to allow for proliferation and differentiation (46).…”

Section: Discussionmentioning

confidence: 99%

Granulosa cells provide transcriptomic information on ovarian follicle dynamics in the southern white rhinoceros

Klohonatz,

Durrant,

Sirard

et al. 2024

Preprint

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Much remains unknown about the reproductive physiology of southern white rhinoceros (SWR) and the effect of ovarian stimulation prior to ovum pickup (OPU) have not been fully elucidated. Granulosa cells (GC) provide valuable insight into follicle growth and oocyte maturation status. The goals of this study were to evaluate transcriptomic changes in GC from three stages of follicle development and to identify biomarkers possibly associated with follicular growth and maturation because of ovarian stimulation. GC collected from SWRs following OPU were assigned stages based upon follicle size. Total RNA was isolated, and cDNA libraries were prepared and sequenced on a NovaSeq 6000. All bioinformatics analyses were performed utilizing the Galaxy web platform. Reads were aligned to CerSimCot1.0, and the manual curation was performed with EquCab3.0. Overall, 39,455 transcripts (21,612 genes) were identified across follicle stages, and manual curation yielded a 61% increase in gene identification from the original annotation. Granulosa cells from preovulatory follicles expressed the highest number of unique transcripts. The following seven biomarkers were determined based upon cluster analysis and patterns of expression: COL1A1, JMY, FBXW11, NRG1, TMPO, MACIR and COL4A1. These data can be used to potentially evaluate the effects of different ovarian stimulation protocols on follicle dynamics, improve OPU results, and support conservation efforts in this species.

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Section: Discussionmentioning

confidence: 99%

Granulosa cells provide transcriptomic information on ovarian follicle dynamics in the southern white rhinoceros

Klohonatz,

Durrant,

Sirard

et al. 2024

Preprint

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“…FOXL2 was considered as a marker of the granulosa cell in mammals and poultry (Du et al, 2022; Luo et al, 2020). In this study, all the granulosa cells isolated and cultured were FOXL2 positive.…”

Section: Discussionmentioning

confidence: 99%

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

Hou,

Hu,

et al. 2023

Journal Cellular Physiology

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Follicle selection in hens refers to a biological process that only one small yellow follicle (SYF) is selected daily or near‐daily for following hierarchical development (from F5/F6 to F1) until ovulation. MFN2 is a kind of GTPases located on the mitochondrial outer membrane, which plays a crucial role in mitochondrial fusion. This study aimed to elucidate the role of MFN2 in proliferation and progesterone biosynthesis of granulosa cells (GCs) during follicle selection in hens. The results showed that GCs began to produce progesterone (P4) after follicle selection, accompanied with changes from multi‐layer with flat cells to single layer with cubic cells. MFN2 was detected in GCs of follicles from SYF to F1. After follicle selection, the expression level of MFN2 in GCs upregulated significantly, accompanied with increases in P4 biosynthesis, ATP production, mitochondrial DNA (mtDNA) copy numbers of granulosa cells. FSH (80 ng/mL) facilitated the effects of P4 biosynthesis and secretion, ATP production, mtDNA copy numbers, cell proliferation and the MFN2 transcription of granulosa cells from F5 (F5G) in vitro. However, FSH treatment did not promote P4 secretion in granulosa cells from SYF (SYFG) in vitro. Meanwhile, we observed that change fold of MFN2 transcription, ATP production, mtDNA copy numbers and cell proliferation rate in F5G after treatment with FSH were greater than those in SYFG. Furthermore, expression levels of MFN2 protein and messenger RNA in F5G were significantly higher than those in SYFG after treatment with FSH. P4 biosynthesis, ATP production, mtDNA copy numbers as well as cell proliferation reduced significantly in F5G with MFN2 knockdown. Oppositely, P4 biosynthesis, ATP production, mtDNA copy numbers and cell proliferation increased significantly in SYFG after the overexpression of MFN2. Our results suggest that the upregulation of MFN2 may be involved in the initiation of P4 biosynthesis, and promotion of GCs proliferation during follicle selection.

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“…It is widely believed that the expression levels of FSHR and cAMP in granulosa cells are important biological markers for follicle selection in laying hens [ 14 ], and the expression level of FSHR mRNA is the highest in 6–8 mm small yellow follicles compared to immature follicles [ 15 ]. Sexual maturation and ovary development are not only regulated by several growth factors, such as growth differentiation factor 9 [ 16 ], Wilms tumor gene 1 [ 17 ], gremlin 1 gene [ 18 ], and forkhead box L2 [ 19 ]. They are also regulated by insulin-like growth factor [ 20 ], epidermal growth factor [ 21 ], and vasoactive intestinal peptide [ 22 ].…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2023

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Sexual maturation provides economically important traits in poultry production. Research on the initiation mechanism of sexual maturity is of great significance for breeding high-yield laying hens. However, the underlying mechanisms are not fully clear. Here, one hundred and fifty Chahua No. 2 laying hens (the CH2 group, which has precocious puberty) and one hundred and fifty Wu Liang Shan black-bone laying hens (the WLS group, a late-maturing chicken breed) with similar weights and ages were randomly selected. ELISA was used to determine the secretion levels of luteinizing hormone (LH), estradiol (E2), and progesterone (P4) in 150-day-old serum and small yellow follicle (SYF) tissues. A histology examination, immunohistochemistry, and quantitative real-time PCR (qPCR) were used to explore the molecular mechanism of how some genes related to oxidative stress affect sexual maturation. The results showed that the secretion levels of LH, E2, and P4 in the CH2 group serum and SYF were higher than those in the WLS group. The results of the real-time PCR of all genes showed that the expression levels of cytochrome P450 family 11 subfamily A member 1, steroidogenic acute regulatory protein, follicle-stimulating hormone receptor, and cytochrome P450 family 19 subfamily A member 1 in the CH2 group were significantly higher than those in the WLS groups (p < 0.001). Untargeted metabolomics combined with multivariate statistical analysis was used to identify biomarkers of SYF tissues in the CH2 and WLS groups. A trajectory analysis of the principal component analysis (PCA) results showed that the samples within the group were clustered and that the samples were dispersed between the CH2 and the WLS groups, indicating that the results of the measured data were reliable and could be used for further research. Further analysis showed that a total of 319 metabolites in small yellow follicles of the CH2 and WLS groups were identified, among which 54 downregulated differential metabolites were identified. These 54 metabolites were found as potential CH2 biomarkers compared with WLS at 150 days, and the different expressions of L-arginine, L-prolinamide, (R)-4-hydroxymandelate, glutathione, and homovanillic acid were more significant. Twenty metabolic pathways were found when significantly differential metabolites were queried in the KEGG database. According to the impact values of the metabolic pathways, eighteen differential metabolites belonged to the mTOR signaling pathway, glutathione metabolism, ABC transporters, the cell ferroptosis pathway, and D-arginine and D-ornithine metabolism. Interestingly, we identified that the cell ferroptosis pathway played an important role in chicken follicle selection for the first time. The histology and immunohistochemistry of SYF showed that the number of granulosa cells increased in the CH2 groups and the expression levels of glutathione peroxidase 4, tumor protein p53, ribosomal protein S6 kinase, and sterol regulatory element binding protein 1 in the granulosa cell layer were upregulated in the CH2 group at the time of sexual maturation. Furthermore, we also speculated that the antioxidant system may play an indispensable role in regulating sexual maturity in chickens. Overall, our findings suggest differentially expressed metabolites and metabolic pathways between CH2 and WLS chickens, providing new insights into the initiation mechanism of sexual maturation.

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FOXL2 regulates the expression of the Col4a1 collagen gene in chicken granulosa cells

Cited by 10 publications

References 21 publications

Granulosa cells provide transcriptomic information on ovarian follicle dynamics in the southern white rhinoceros

Granulosa cells provide transcriptomic information on ovarian follicle dynamics in the southern white rhinoceros

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

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

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