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PROP1 triggers epithelial-mesenchymal transition-like process in pituitary stem cells

View Article: PubMed Central - PubMed

ABSTRACT

Mutations in PROP1 are the most common cause of hypopituitarism in humans; therefore, unraveling its mechanism of action is highly relevant from a therapeutic perspective. Our current understanding of the role of PROP1 in the pituitary gland is limited to the repression and activation of the pituitary transcription factor genes Hesx1 and Pou1f1, respectively. To elucidate the comprehensive PROP1-dependent gene regulatory network, we conducted genome-wide analysis of PROP1 DNA binding and effects on gene expression in mutant mice, mouse isolated stem cells and engineered mouse cell lines. We determined that PROP1 is essential for stimulating stem cells to undergo an epithelial to mesenchymal transition-like process necessary for cell migration and differentiation. Genomic profiling reveals that PROP1 binds to genes expressed in epithelial cells like Claudin 23, and to EMT inducer genes like Zeb2, Notch2 and Gli2. Zeb2 activation appears to be a key step in the EMT process. Our findings identify PROP1 as a central transcriptional component of pituitary stem cell differentiation.

Doi:: http://dx.doi.org/10.7554/eLife.14470.001

No MeSH data available.


Related in: MedlinePlus

Temporal expression patterns of Prop1, PROP1 targets and stem cell markers during pituitary development.Relative mRNA levels of Prop1, Notch2, Pit1, Sox2, Gfra2 and Sox9 in pituitaries of wild-type mice (n=7) were determined by semi-quantitative real-time PCR at different time points (e12.5, e14.5, e18.5 and postnatal day 3 and 7). ANOVA, post hoc Fisher analysis: letters and *show a statistically significant difference, p<0.05. Samples were normalized to GAPDH mRNA. Plots denote the mean ± SEM.DOI:http://dx.doi.org/10.7554/eLife.14470.004
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fig2s1: Temporal expression patterns of Prop1, PROP1 targets and stem cell markers during pituitary development.Relative mRNA levels of Prop1, Notch2, Pit1, Sox2, Gfra2 and Sox9 in pituitaries of wild-type mice (n=7) were determined by semi-quantitative real-time PCR at different time points (e12.5, e14.5, e18.5 and postnatal day 3 and 7). ANOVA, post hoc Fisher analysis: letters and *show a statistically significant difference, p<0.05. Samples were normalized to GAPDH mRNA. Plots denote the mean ± SEM.DOI:http://dx.doi.org/10.7554/eLife.14470.004

Mentions: The rodent pituitary gland undergoes two distinct waves of cell proliferation and differentiation, one occurring during embryogenesis and a second one during the first 3 weeks afterbirth in the mouse (Gremeaux et al., 2012; Zhu et al., 2007; Carbajo-Pérez and Watanabe, 1990). The known pattern of Prop1 expression correlates with the first wave of proliferation, which peaks at e12.5 and wanes at e14.5, but expression during the postnatal wave of cell proliferation has not been investigated. Using qRT-PCR, we discovered high Prop1 mRNA levels at postnatal days 3 and 7 (P3 and P7), that are similar to the peak levels at e12.5 and coincident with the second wave of cell proliferation (Figure 2—figure supplement 1). We also used qRT-PCR to assess the temporal expression patterns of Prop1-dependent genes (Pou1f1 and Notch2), and the stem cell markers Sox2, Gfra2 and Sox9 during these waves of pituitary growth (Figure 2—figure supplement 1). We found that Prop1 and all these genes are expressed during the postnatal wave of pituitary expansion, and their mRNA levels are at higher or similar levels to those found in embryonic pituitaries.


PROP1 triggers epithelial-mesenchymal transition-like process in pituitary stem cells
Temporal expression patterns of Prop1, PROP1 targets and stem cell markers during pituitary development.Relative mRNA levels of Prop1, Notch2, Pit1, Sox2, Gfra2 and Sox9 in pituitaries of wild-type mice (n=7) were determined by semi-quantitative real-time PCR at different time points (e12.5, e14.5, e18.5 and postnatal day 3 and 7). ANOVA, post hoc Fisher analysis: letters and *show a statistically significant difference, p<0.05. Samples were normalized to GAPDH mRNA. Plots denote the mean ± SEM.DOI:http://dx.doi.org/10.7554/eLife.14470.004
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4940164&req=5

fig2s1: Temporal expression patterns of Prop1, PROP1 targets and stem cell markers during pituitary development.Relative mRNA levels of Prop1, Notch2, Pit1, Sox2, Gfra2 and Sox9 in pituitaries of wild-type mice (n=7) were determined by semi-quantitative real-time PCR at different time points (e12.5, e14.5, e18.5 and postnatal day 3 and 7). ANOVA, post hoc Fisher analysis: letters and *show a statistically significant difference, p<0.05. Samples were normalized to GAPDH mRNA. Plots denote the mean ± SEM.DOI:http://dx.doi.org/10.7554/eLife.14470.004
Mentions: The rodent pituitary gland undergoes two distinct waves of cell proliferation and differentiation, one occurring during embryogenesis and a second one during the first 3 weeks afterbirth in the mouse (Gremeaux et al., 2012; Zhu et al., 2007; Carbajo-Pérez and Watanabe, 1990). The known pattern of Prop1 expression correlates with the first wave of proliferation, which peaks at e12.5 and wanes at e14.5, but expression during the postnatal wave of cell proliferation has not been investigated. Using qRT-PCR, we discovered high Prop1 mRNA levels at postnatal days 3 and 7 (P3 and P7), that are similar to the peak levels at e12.5 and coincident with the second wave of cell proliferation (Figure 2—figure supplement 1). We also used qRT-PCR to assess the temporal expression patterns of Prop1-dependent genes (Pou1f1 and Notch2), and the stem cell markers Sox2, Gfra2 and Sox9 during these waves of pituitary growth (Figure 2—figure supplement 1). We found that Prop1 and all these genes are expressed during the postnatal wave of pituitary expansion, and their mRNA levels are at higher or similar levels to those found in embryonic pituitaries.

View Article: PubMed Central - PubMed

ABSTRACT

Mutations in PROP1 are the most common cause of hypopituitarism in humans; therefore, unraveling its mechanism of action is highly relevant from a therapeutic perspective. Our current understanding of the role of PROP1 in the pituitary gland is limited to the repression and activation of the pituitary transcription factor genes Hesx1 and Pou1f1, respectively. To elucidate the comprehensive PROP1-dependent gene regulatory network, we conducted genome-wide analysis of PROP1 DNA binding and effects on gene expression in mutant mice, mouse isolated stem cells and engineered mouse cell lines. We determined that PROP1 is essential for stimulating stem cells to undergo an epithelial to mesenchymal transition-like process necessary for cell migration and differentiation. Genomic profiling reveals that PROP1 binds to genes expressed in epithelial cells like Claudin 23, and to EMT inducer genes like Zeb2, Notch2 and Gli2. Zeb2 activation appears to be a key step in the EMT process. Our findings identify PROP1 as a central transcriptional component of pituitary stem cell differentiation.

Doi:: http://dx.doi.org/10.7554/eLife.14470.001

No MeSH data available.


Related in: MedlinePlus