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Regulation and regulatory role of WNT signaling in potentiating FSH action during bovine dominant follicle selection.

Gupta PS, Folger JK, Rajput SK, Lv L, Yao J, Ireland JJ, Smith GW - PLoS ONE (2014)

Bottom Line: FSH treatment increased CYP19A1, CCND2, CTNNB1, AXIN2 and FZD6 mRNAs and the stimulatory effect on CYP19A1 mRNA was reduced by IWR-1.In contrast, FSH reduced CARTPT mRNA and IWR-1 partially reversed the inhibitory effect of FSH.Results support temporal and hormonal regulation and a potential role for WNT signaling in potentiating FSH action during dominant follicle selection.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Mammalian Reproductive Biology and Genomics, Michigan State University, East Lansing, Michigan, United States of America; Department of Animal Science, Michigan State University, East Lansing, Michigan, United States of America.

ABSTRACT
Follicular development occurs in wave like patterns in monotocous species such as cattle and humans and is regulated by a complex interaction of gonadotropins with local intrafollicular regulatory molecules. To further elucidate potential mechanisms controlling dominant follicle selection, granulosa cell RNA harvested from F1 (largest) and F2 (second largest) follicles isolated at predeviation (PD) and onset of diameter deviation (OD) stages of the first follicular wave was subjected to preliminary RNA transcriptome analysis. Expression of numerous WNT system components was observed. Hence experiments were performed to test the hypothesis that WNT signaling modulates FSH action on granulosa cells during follicular waves. Abundance of mRNA for WNT pathway members was evaluated in granulosa cells harvested from follicles at emergence (EM), PD, OD and early dominance (ED) stages of the first follicular wave. In F1 follicles, abundance of CTNNB1 and DVL1 mRNAs was higher and AXIN2 mRNA was lower at ED versus EM stages and DVL1 and FZD6 mRNAs were higher and AXIN2 mRNA was lower in F1 versus F2 follicle at the ED stage. Bovine granulosa cells were treated in vitro with increasing doses of the WNT inhibitor IWR-1+/- maximal stimulatory dose of FSH. IWR-1 treatment blocked the FSH-induced increase in granulosa cell numbers and reduced the FSH-induced increase in estradiol. Granulosa cells were also cultured in the presence or absence of FSH +/- IWR-1 and hormonal regulation of mRNA for WNT pathway members and known FSH targets determined. FSH treatment increased CYP19A1, CCND2, CTNNB1, AXIN2 and FZD6 mRNAs and the stimulatory effect on CYP19A1 mRNA was reduced by IWR-1. In contrast, FSH reduced CARTPT mRNA and IWR-1 partially reversed the inhibitory effect of FSH. Results support temporal and hormonal regulation and a potential role for WNT signaling in potentiating FSH action during dominant follicle selection.

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Effect of FSH and WNT signaling inhibitor (IWR-1) treatment on FZD6, AXIN2, CTNNB1 and DVL1 mRNA abundance in bovine granulosa cells.Quantitative RT-PCR analysis was performed to determine the effects of treatment with 0 or 0.5 ng/ml FSH in the presence or absence of 1.0 µM IWR-1 on FZD6 (A), CCND2 (B) and CARTPT (C) mRNA expression in cultured granulosa cells. Expression of mRNA for genes of interest was normalized relative to abundance of mRNA for ACTIN as internal control. Bars represent mean +/− SEM for four experiments. A,B P<0.05.
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pone-0100201-g005: Effect of FSH and WNT signaling inhibitor (IWR-1) treatment on FZD6, AXIN2, CTNNB1 and DVL1 mRNA abundance in bovine granulosa cells.Quantitative RT-PCR analysis was performed to determine the effects of treatment with 0 or 0.5 ng/ml FSH in the presence or absence of 1.0 µM IWR-1 on FZD6 (A), CCND2 (B) and CARTPT (C) mRNA expression in cultured granulosa cells. Expression of mRNA for genes of interest was normalized relative to abundance of mRNA for ACTIN as internal control. Bars represent mean +/− SEM for four experiments. A,B P<0.05.

Mentions: Above results support a prominent role for WNT signaling in potentiating specific components of FSH action in bovine granulosa cells. To further understand the potential functional role in follicular selection, the hormonal regulation of expression of select WNT system components in bovine granulosa cells was investigated. Granulosa cells cultured in the presence of FSH exhibited increased expression of AXIN2, CTNNB1 and FZD6 mRNA (Figure 5A-C) compared to untreated controls (P<0.05), but no effect of inhibition of WNT signaling via IWR-1 treatment on basal and FSH-induced mRNA expression for AXIN2, CTNNB1 and FZD6 was observed. In contrast mRNA abundance for DVL1 was not impacted by FSH or IWR-1 treatment (Figure 5D). Results support WNT-CTNNB1 independent FSH-induced regulation of expression of specific WNT system components in bovine granulosa cells.


Regulation and regulatory role of WNT signaling in potentiating FSH action during bovine dominant follicle selection.

Gupta PS, Folger JK, Rajput SK, Lv L, Yao J, Ireland JJ, Smith GW - PLoS ONE (2014)

Effect of FSH and WNT signaling inhibitor (IWR-1) treatment on FZD6, AXIN2, CTNNB1 and DVL1 mRNA abundance in bovine granulosa cells.Quantitative RT-PCR analysis was performed to determine the effects of treatment with 0 or 0.5 ng/ml FSH in the presence or absence of 1.0 µM IWR-1 on FZD6 (A), CCND2 (B) and CARTPT (C) mRNA expression in cultured granulosa cells. Expression of mRNA for genes of interest was normalized relative to abundance of mRNA for ACTIN as internal control. Bars represent mean +/− SEM for four experiments. A,B P<0.05.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4061082&req=5

pone-0100201-g005: Effect of FSH and WNT signaling inhibitor (IWR-1) treatment on FZD6, AXIN2, CTNNB1 and DVL1 mRNA abundance in bovine granulosa cells.Quantitative RT-PCR analysis was performed to determine the effects of treatment with 0 or 0.5 ng/ml FSH in the presence or absence of 1.0 µM IWR-1 on FZD6 (A), CCND2 (B) and CARTPT (C) mRNA expression in cultured granulosa cells. Expression of mRNA for genes of interest was normalized relative to abundance of mRNA for ACTIN as internal control. Bars represent mean +/− SEM for four experiments. A,B P<0.05.
Mentions: Above results support a prominent role for WNT signaling in potentiating specific components of FSH action in bovine granulosa cells. To further understand the potential functional role in follicular selection, the hormonal regulation of expression of select WNT system components in bovine granulosa cells was investigated. Granulosa cells cultured in the presence of FSH exhibited increased expression of AXIN2, CTNNB1 and FZD6 mRNA (Figure 5A-C) compared to untreated controls (P<0.05), but no effect of inhibition of WNT signaling via IWR-1 treatment on basal and FSH-induced mRNA expression for AXIN2, CTNNB1 and FZD6 was observed. In contrast mRNA abundance for DVL1 was not impacted by FSH or IWR-1 treatment (Figure 5D). Results support WNT-CTNNB1 independent FSH-induced regulation of expression of specific WNT system components in bovine granulosa cells.

Bottom Line: FSH treatment increased CYP19A1, CCND2, CTNNB1, AXIN2 and FZD6 mRNAs and the stimulatory effect on CYP19A1 mRNA was reduced by IWR-1.In contrast, FSH reduced CARTPT mRNA and IWR-1 partially reversed the inhibitory effect of FSH.Results support temporal and hormonal regulation and a potential role for WNT signaling in potentiating FSH action during dominant follicle selection.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Mammalian Reproductive Biology and Genomics, Michigan State University, East Lansing, Michigan, United States of America; Department of Animal Science, Michigan State University, East Lansing, Michigan, United States of America.

ABSTRACT
Follicular development occurs in wave like patterns in monotocous species such as cattle and humans and is regulated by a complex interaction of gonadotropins with local intrafollicular regulatory molecules. To further elucidate potential mechanisms controlling dominant follicle selection, granulosa cell RNA harvested from F1 (largest) and F2 (second largest) follicles isolated at predeviation (PD) and onset of diameter deviation (OD) stages of the first follicular wave was subjected to preliminary RNA transcriptome analysis. Expression of numerous WNT system components was observed. Hence experiments were performed to test the hypothesis that WNT signaling modulates FSH action on granulosa cells during follicular waves. Abundance of mRNA for WNT pathway members was evaluated in granulosa cells harvested from follicles at emergence (EM), PD, OD and early dominance (ED) stages of the first follicular wave. In F1 follicles, abundance of CTNNB1 and DVL1 mRNAs was higher and AXIN2 mRNA was lower at ED versus EM stages and DVL1 and FZD6 mRNAs were higher and AXIN2 mRNA was lower in F1 versus F2 follicle at the ED stage. Bovine granulosa cells were treated in vitro with increasing doses of the WNT inhibitor IWR-1+/- maximal stimulatory dose of FSH. IWR-1 treatment blocked the FSH-induced increase in granulosa cell numbers and reduced the FSH-induced increase in estradiol. Granulosa cells were also cultured in the presence or absence of FSH +/- IWR-1 and hormonal regulation of mRNA for WNT pathway members and known FSH targets determined. FSH treatment increased CYP19A1, CCND2, CTNNB1, AXIN2 and FZD6 mRNAs and the stimulatory effect on CYP19A1 mRNA was reduced by IWR-1. In contrast, FSH reduced CARTPT mRNA and IWR-1 partially reversed the inhibitory effect of FSH. Results support temporal and hormonal regulation and a potential role for WNT signaling in potentiating FSH action during dominant follicle selection.

Show MeSH
Related in: MedlinePlus