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Ornithine Decarboxylase Activity Is Required for Prostatic Budding in the Developing Mouse Prostate.

Gamat M, Malinowski RL, Parkhurst LJ, Steinke LM, Marker PC - PLoS ONE (2015)

Bottom Line: Inhibiting ornithine decarboxylase using DFMO in UGS organ culture blocked the induction of prostatic buds by androgens, and significantly decreased expression of key prostate transcription factor, Nkx3.1, by androgens.DFMO also significantly decreased the expression of developmental regulatory gene Notch1.Together these results indicate that Odc1 and polyamines are required for androgens to exert their effect in mediating prostatic bud induction, and are required for the expression of a subset of prostatic developmental regulatory genes including Notch1 and Nkx3.1.

View Article: PubMed Central - PubMed

Affiliation: School of Pharmacy, University of Wisconsin-Madison, 777 Highland Avenue, Madison, WI, United States of America.

ABSTRACT
The prostate is a male accessory sex gland that produces secretions in seminal fluid to facilitate fertilization. Prostate secretory function is dependent on androgens, although the mechanism by which androgens exert their effects is still unclear. Polyamines are small cationic molecules that play pivotal roles in DNA transcription, translation and gene regulation. The rate-limiting enzyme in polyamine biosynthesis is ornithine decarboxylase, which is encoded by the gene Odc1. Ornithine decarboxylase mRNA decreases in the prostate upon castration and increases upon administration of androgens. Furthermore, testosterone administered to castrated male mice restores prostate secretory activity, whereas administering testosterone and the ornithine decarboxylase inhibitor D,L-α-difluromethylornithine (DFMO) to castrated males does not restore prostate secretory activity, suggesting that polyamines are required for androgens to exert their effects. To date, no one has examined polyamines in prostate development, which is also androgen dependent. In this study, we showed that ornithine decarboxylase protein was expressed in the epithelium of the ventral, dorsolateral and anterior lobes of the adult mouse prostate. Ornithine decarboxylase protein was also expressed in the urogenital sinus (UGS) epithelium of the male and female embryo prior to prostate development, and expression continued in prostatic epithelial buds as they emerged from the UGS. Inhibiting ornithine decarboxylase using DFMO in UGS organ culture blocked the induction of prostatic buds by androgens, and significantly decreased expression of key prostate transcription factor, Nkx3.1, by androgens. DFMO also significantly decreased the expression of developmental regulatory gene Notch1. Other genes implicated in prostatic development including Sox9, Wif1 and Srd5a2 were unaffected by DFMO. Together these results indicate that Odc1 and polyamines are required for androgens to exert their effect in mediating prostatic bud induction, and are required for the expression of a subset of prostatic developmental regulatory genes including Notch1 and Nkx3.1.

No MeSH data available.


Related in: MedlinePlus

Markers of tissue compartments in the cultured UGS.Cytokeratin staining was confined to cytoplasm of the urogenital epithelial cells in the NS control (A), T treated UGS (B) and in the T+DFMO treated UGS (C). p63 staining was present in the nuclei of epithelial cells in the NS control (D), T treated UGS (E) and in the T+DFMO treated UGS (F). SMA staining was present in the cytoplasm of peri-ductal mesenchymal cells in the NS control (G), T treated UGS (H) and T+DFMO treated UGS (I). Positive immunostaining is brown. The dotted line demarcates the boundary between epithelium and mesenchyme. Abbreviations: B prostatic bud, cyto cytokeratin, DFMO difluoromethylornithine, E epithelium, M mesenchyme, NS no steroid control, SMA smooth muscle actin, T testosterone.
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pone.0139522.g006: Markers of tissue compartments in the cultured UGS.Cytokeratin staining was confined to cytoplasm of the urogenital epithelial cells in the NS control (A), T treated UGS (B) and in the T+DFMO treated UGS (C). p63 staining was present in the nuclei of epithelial cells in the NS control (D), T treated UGS (E) and in the T+DFMO treated UGS (F). SMA staining was present in the cytoplasm of peri-ductal mesenchymal cells in the NS control (G), T treated UGS (H) and T+DFMO treated UGS (I). Positive immunostaining is brown. The dotted line demarcates the boundary between epithelium and mesenchyme. Abbreviations: B prostatic bud, cyto cytokeratin, DFMO difluoromethylornithine, E epithelium, M mesenchyme, NS no steroid control, SMA smooth muscle actin, T testosterone.

Mentions: Using immunohistochemistry, we showed that the urogenital epithelium expressed cytokeratins. In the NS treated UGS, cytokeratins were expressed in the urogenital epithelium, which remained smooth after six days in culture media (Fig 6A). In the testosterone treated UGS, cytokeratins were expressed in all the cells of the urogenital epithelium, which underwent extensive bud formation (Fig 6B). In the T+DFMO treated UGS, cytokeratins were still expressed in the epithelium, which did not undergo prostatic budding (Fig 6C). Under all conditions, cytokeratins were always expressed in the epithelium, suggesting that polyamines did not alter cytokeratin expression in epithelial cells.


Ornithine Decarboxylase Activity Is Required for Prostatic Budding in the Developing Mouse Prostate.

Gamat M, Malinowski RL, Parkhurst LJ, Steinke LM, Marker PC - PLoS ONE (2015)

Markers of tissue compartments in the cultured UGS.Cytokeratin staining was confined to cytoplasm of the urogenital epithelial cells in the NS control (A), T treated UGS (B) and in the T+DFMO treated UGS (C). p63 staining was present in the nuclei of epithelial cells in the NS control (D), T treated UGS (E) and in the T+DFMO treated UGS (F). SMA staining was present in the cytoplasm of peri-ductal mesenchymal cells in the NS control (G), T treated UGS (H) and T+DFMO treated UGS (I). Positive immunostaining is brown. The dotted line demarcates the boundary between epithelium and mesenchyme. Abbreviations: B prostatic bud, cyto cytokeratin, DFMO difluoromethylornithine, E epithelium, M mesenchyme, NS no steroid control, SMA smooth muscle actin, T testosterone.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591331&req=5

pone.0139522.g006: Markers of tissue compartments in the cultured UGS.Cytokeratin staining was confined to cytoplasm of the urogenital epithelial cells in the NS control (A), T treated UGS (B) and in the T+DFMO treated UGS (C). p63 staining was present in the nuclei of epithelial cells in the NS control (D), T treated UGS (E) and in the T+DFMO treated UGS (F). SMA staining was present in the cytoplasm of peri-ductal mesenchymal cells in the NS control (G), T treated UGS (H) and T+DFMO treated UGS (I). Positive immunostaining is brown. The dotted line demarcates the boundary between epithelium and mesenchyme. Abbreviations: B prostatic bud, cyto cytokeratin, DFMO difluoromethylornithine, E epithelium, M mesenchyme, NS no steroid control, SMA smooth muscle actin, T testosterone.
Mentions: Using immunohistochemistry, we showed that the urogenital epithelium expressed cytokeratins. In the NS treated UGS, cytokeratins were expressed in the urogenital epithelium, which remained smooth after six days in culture media (Fig 6A). In the testosterone treated UGS, cytokeratins were expressed in all the cells of the urogenital epithelium, which underwent extensive bud formation (Fig 6B). In the T+DFMO treated UGS, cytokeratins were still expressed in the epithelium, which did not undergo prostatic budding (Fig 6C). Under all conditions, cytokeratins were always expressed in the epithelium, suggesting that polyamines did not alter cytokeratin expression in epithelial cells.

Bottom Line: Inhibiting ornithine decarboxylase using DFMO in UGS organ culture blocked the induction of prostatic buds by androgens, and significantly decreased expression of key prostate transcription factor, Nkx3.1, by androgens.DFMO also significantly decreased the expression of developmental regulatory gene Notch1.Together these results indicate that Odc1 and polyamines are required for androgens to exert their effect in mediating prostatic bud induction, and are required for the expression of a subset of prostatic developmental regulatory genes including Notch1 and Nkx3.1.

View Article: PubMed Central - PubMed

Affiliation: School of Pharmacy, University of Wisconsin-Madison, 777 Highland Avenue, Madison, WI, United States of America.

ABSTRACT
The prostate is a male accessory sex gland that produces secretions in seminal fluid to facilitate fertilization. Prostate secretory function is dependent on androgens, although the mechanism by which androgens exert their effects is still unclear. Polyamines are small cationic molecules that play pivotal roles in DNA transcription, translation and gene regulation. The rate-limiting enzyme in polyamine biosynthesis is ornithine decarboxylase, which is encoded by the gene Odc1. Ornithine decarboxylase mRNA decreases in the prostate upon castration and increases upon administration of androgens. Furthermore, testosterone administered to castrated male mice restores prostate secretory activity, whereas administering testosterone and the ornithine decarboxylase inhibitor D,L-α-difluromethylornithine (DFMO) to castrated males does not restore prostate secretory activity, suggesting that polyamines are required for androgens to exert their effects. To date, no one has examined polyamines in prostate development, which is also androgen dependent. In this study, we showed that ornithine decarboxylase protein was expressed in the epithelium of the ventral, dorsolateral and anterior lobes of the adult mouse prostate. Ornithine decarboxylase protein was also expressed in the urogenital sinus (UGS) epithelium of the male and female embryo prior to prostate development, and expression continued in prostatic epithelial buds as they emerged from the UGS. Inhibiting ornithine decarboxylase using DFMO in UGS organ culture blocked the induction of prostatic buds by androgens, and significantly decreased expression of key prostate transcription factor, Nkx3.1, by androgens. DFMO also significantly decreased the expression of developmental regulatory gene Notch1. Other genes implicated in prostatic development including Sox9, Wif1 and Srd5a2 were unaffected by DFMO. Together these results indicate that Odc1 and polyamines are required for androgens to exert their effect in mediating prostatic bud induction, and are required for the expression of a subset of prostatic developmental regulatory genes including Notch1 and Nkx3.1.

No MeSH data available.


Related in: MedlinePlus