Limits...
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

Ornithine decarboxylase protein was present in the urogenital epithelium in the developing urogenital sinus.In the male UGS before budding at E15 (A) and E16 (B), ornithine decarboxylase protein (green) was present in the epithelium. As the buds emerged from the epithelium at E17 (E) and E18 (G), ornithine decarboxylase protein was still expressed in the epithelium and buds. In the female UGS, ornithine decarboxylase was present in the epithelium at E15 (B) and E16 (D), the period before prostatic buds are initiated in the male UGS. At E17 (F) and E18 (H), ornithine decarboxylase protein continued to be expressed in the urogenital epithelium. Ornithine decarboxylase staining is in green, and vimentin staining is in red. There was some co-localization of ornithine decarboxylase and vimentin (yellow) in the mesenchyme. Arrowheads denote prostatic buds. Abbreviations: E epithelium, M mesenchyme.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4591331&req=5

pone.0139522.g003: Ornithine decarboxylase protein was present in the urogenital epithelium in the developing urogenital sinus.In the male UGS before budding at E15 (A) and E16 (B), ornithine decarboxylase protein (green) was present in the epithelium. As the buds emerged from the epithelium at E17 (E) and E18 (G), ornithine decarboxylase protein was still expressed in the epithelium and buds. In the female UGS, ornithine decarboxylase was present in the epithelium at E15 (B) and E16 (D), the period before prostatic buds are initiated in the male UGS. At E17 (F) and E18 (H), ornithine decarboxylase protein continued to be expressed in the urogenital epithelium. Ornithine decarboxylase staining is in green, and vimentin staining is in red. There was some co-localization of ornithine decarboxylase and vimentin (yellow) in the mesenchyme. Arrowheads denote prostatic buds. Abbreviations: E epithelium, M mesenchyme.

Mentions: We then assessed the expression of ornithine decarboxylase protein in the developing UGS using immunofluorescence, and compared male and female tissues at similar stages. We used vimentin to distinguish mesenchymal tissue (red) from the urogenital epithelium (Fig 3). Ornithine decarboxylase protein (green) was expressed in the UGS in both the male (Fig 3A, 3C, 3E and 3G) and the female (Fig 3B, 3D, 3F and 3H). In the male UGS, ornithine decarboxylase protein was present in the urogenital epithelium before budding was apparent at E15 (Fig 3A) and E16 (Fig 3C). By E17, prostatic epithelial buds were apparent, and projecting into the mesenchyme (Fig 3E, arrows), and ornithine decarboxylase staining was expressed in the urogenital epithlium as well as the buds. By E18, ornithine decarboxylase protein was still expressed in the urogenital and developing prostatic epithelium (Fig 3G, arrows). In the female UGS, the epithelium remained smooth with no apparent signs of prostatic budding (Fig 3B, 3D, 3F and 3G). Similar to the male UGS, ornithine decarboxylase protein was expressed in the urogenital epithelium (Fig 3B, 3D, 3F and 3G). In all stages, there was some ornithine decarboxylase staining co-localized with vimentin (yellow), indicating that there was some ornithine decarboxylase in the mesenchyme. The negative control (with primary antibody omitted) showed no staining (data not shown).


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)

Ornithine decarboxylase protein was present in the urogenital epithelium in the developing urogenital sinus.In the male UGS before budding at E15 (A) and E16 (B), ornithine decarboxylase protein (green) was present in the epithelium. As the buds emerged from the epithelium at E17 (E) and E18 (G), ornithine decarboxylase protein was still expressed in the epithelium and buds. In the female UGS, ornithine decarboxylase was present in the epithelium at E15 (B) and E16 (D), the period before prostatic buds are initiated in the male UGS. At E17 (F) and E18 (H), ornithine decarboxylase protein continued to be expressed in the urogenital epithelium. Ornithine decarboxylase staining is in green, and vimentin staining is in red. There was some co-localization of ornithine decarboxylase and vimentin (yellow) in the mesenchyme. Arrowheads denote prostatic buds. Abbreviations: E epithelium, M mesenchyme.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0139522.g003: Ornithine decarboxylase protein was present in the urogenital epithelium in the developing urogenital sinus.In the male UGS before budding at E15 (A) and E16 (B), ornithine decarboxylase protein (green) was present in the epithelium. As the buds emerged from the epithelium at E17 (E) and E18 (G), ornithine decarboxylase protein was still expressed in the epithelium and buds. In the female UGS, ornithine decarboxylase was present in the epithelium at E15 (B) and E16 (D), the period before prostatic buds are initiated in the male UGS. At E17 (F) and E18 (H), ornithine decarboxylase protein continued to be expressed in the urogenital epithelium. Ornithine decarboxylase staining is in green, and vimentin staining is in red. There was some co-localization of ornithine decarboxylase and vimentin (yellow) in the mesenchyme. Arrowheads denote prostatic buds. Abbreviations: E epithelium, M mesenchyme.
Mentions: We then assessed the expression of ornithine decarboxylase protein in the developing UGS using immunofluorescence, and compared male and female tissues at similar stages. We used vimentin to distinguish mesenchymal tissue (red) from the urogenital epithelium (Fig 3). Ornithine decarboxylase protein (green) was expressed in the UGS in both the male (Fig 3A, 3C, 3E and 3G) and the female (Fig 3B, 3D, 3F and 3H). In the male UGS, ornithine decarboxylase protein was present in the urogenital epithelium before budding was apparent at E15 (Fig 3A) and E16 (Fig 3C). By E17, prostatic epithelial buds were apparent, and projecting into the mesenchyme (Fig 3E, arrows), and ornithine decarboxylase staining was expressed in the urogenital epithlium as well as the buds. By E18, ornithine decarboxylase protein was still expressed in the urogenital and developing prostatic epithelium (Fig 3G, arrows). In the female UGS, the epithelium remained smooth with no apparent signs of prostatic budding (Fig 3B, 3D, 3F and 3G). Similar to the male UGS, ornithine decarboxylase protein was expressed in the urogenital epithelium (Fig 3B, 3D, 3F and 3G). In all stages, there was some ornithine decarboxylase staining co-localized with vimentin (yellow), indicating that there was some ornithine decarboxylase in the mesenchyme. The negative control (with primary antibody omitted) showed no staining (data not shown).

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