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Correlation between urothelial differentiation and sensory proteins P2X3, P2X5, TRPV1, and TRPV4 in normal urothelium and papillary carcinoma of human bladder.

Sterle I, Zupančič D, Romih R - Biomed Res Int (2014)

Bottom Line: Terminal differentiation of urothelium is a prerequisite for blood-urine barrier formation and enables normal sensory function of the urinary bladder.In this study, urothelial differentiation of normal human urothelium and of low and high grade papillary urothelial carcinomas was correlated with the expression and localization of purinergic receptors (P2X3, and P2X5) and transient receptor potential vanilloid channels (TRPV1, and TRPV4).Our results suggest that sensory proteins P2X3 and TRPV1 are in correlation with urothelial differentiation, while P2X5 and TRPV4 have unique expression patterns.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, University Medical Centre Ljubljana, Zaloška cesta 2, 1000 Ljubljana, Slovenia.

ABSTRACT
Terminal differentiation of urothelium is a prerequisite for blood-urine barrier formation and enables normal sensory function of the urinary bladder. In this study, urothelial differentiation of normal human urothelium and of low and high grade papillary urothelial carcinomas was correlated with the expression and localization of purinergic receptors (P2X3, and P2X5) and transient receptor potential vanilloid channels (TRPV1, and TRPV4). Western blotting and immunofluorescence of uroplakins together with scanning electron microscopy of urothelial apical surface demonstrated terminal differentiation of normal urothelium, partial differentiation of low grade carcinoma, and poor differentiation of high grade carcinoma. P2X3 was expressed in normal urothelium as well as in low grade carcinoma and in both cases immunolabeling was stronger in the superficial cells. P2X3 expression decreased in high grade carcinoma. P2X5 expression was detected in normal urothelium and in high grade carcinoma, while in low grade carcinoma its expression was diminished. The expression of TRPV1 decreased in low grade and even more in high grade carcinoma when compared with normal urothelium, while TRPV4 expression was unchanged in all samples. Our results suggest that sensory proteins P2X3 and TRPV1 are in correlation with urothelial differentiation, while P2X5 and TRPV4 have unique expression patterns.

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High grade papillary urothelial carcinomas (pT1 or pT2). (a) Uroplakin (UP) labelling (red) is negative. (b) Scanning electron microscopy (SEM) shows that superficial urothelial cells are small and polymorphic. They have microvilli on their apical surface. (c) Antibody against P2X3 weakly labels (green) urothelial cells. (d) P2X5 labelling (red) is present in all urothelial cells. Nuclei of some cells are also labelled. (e) TRPV1 labelling is negative. (f) Weak labelling of TRPV4 in urothelial cells (U) is seen, but strong TRPV4 labelling (arrows) is seen in the compartments of the lamina propria (LP). In images (a) and (c)–(f), nuclei are labelled blue with DAPI. L = lumen. Scale bars = 50 μm.
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fig4: High grade papillary urothelial carcinomas (pT1 or pT2). (a) Uroplakin (UP) labelling (red) is negative. (b) Scanning electron microscopy (SEM) shows that superficial urothelial cells are small and polymorphic. They have microvilli on their apical surface. (c) Antibody against P2X3 weakly labels (green) urothelial cells. (d) P2X5 labelling (red) is present in all urothelial cells. Nuclei of some cells are also labelled. (e) TRPV1 labelling is negative. (f) Weak labelling of TRPV4 in urothelial cells (U) is seen, but strong TRPV4 labelling (arrows) is seen in the compartments of the lamina propria (LP). In images (a) and (c)–(f), nuclei are labelled blue with DAPI. L = lumen. Scale bars = 50 μm.

Mentions: We did not observe any difference between pT1 and pT2 high grade papillary urothelial carcinoma with respect to the protein expression and localization studied here. Western blotting showed that there was no uroplakin expression in these samples (Figure 1) and uroplakin immunolabeling was also negative in all samples of pT1 and pT2 (Figure 4(a)). Scanning electron microscopy revealed superficial cells of different sizes, but prevailing ones were smaller than in normal urothelium (Figure 4(b)). They were covered with microvilli (Figure 4(b)), which are found only on poorly differentiated superficial urothelial cells [6].


Correlation between urothelial differentiation and sensory proteins P2X3, P2X5, TRPV1, and TRPV4 in normal urothelium and papillary carcinoma of human bladder.

Sterle I, Zupančič D, Romih R - Biomed Res Int (2014)

High grade papillary urothelial carcinomas (pT1 or pT2). (a) Uroplakin (UP) labelling (red) is negative. (b) Scanning electron microscopy (SEM) shows that superficial urothelial cells are small and polymorphic. They have microvilli on their apical surface. (c) Antibody against P2X3 weakly labels (green) urothelial cells. (d) P2X5 labelling (red) is present in all urothelial cells. Nuclei of some cells are also labelled. (e) TRPV1 labelling is negative. (f) Weak labelling of TRPV4 in urothelial cells (U) is seen, but strong TRPV4 labelling (arrows) is seen in the compartments of the lamina propria (LP). In images (a) and (c)–(f), nuclei are labelled blue with DAPI. L = lumen. Scale bars = 50 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig4: High grade papillary urothelial carcinomas (pT1 or pT2). (a) Uroplakin (UP) labelling (red) is negative. (b) Scanning electron microscopy (SEM) shows that superficial urothelial cells are small and polymorphic. They have microvilli on their apical surface. (c) Antibody against P2X3 weakly labels (green) urothelial cells. (d) P2X5 labelling (red) is present in all urothelial cells. Nuclei of some cells are also labelled. (e) TRPV1 labelling is negative. (f) Weak labelling of TRPV4 in urothelial cells (U) is seen, but strong TRPV4 labelling (arrows) is seen in the compartments of the lamina propria (LP). In images (a) and (c)–(f), nuclei are labelled blue with DAPI. L = lumen. Scale bars = 50 μm.
Mentions: We did not observe any difference between pT1 and pT2 high grade papillary urothelial carcinoma with respect to the protein expression and localization studied here. Western blotting showed that there was no uroplakin expression in these samples (Figure 1) and uroplakin immunolabeling was also negative in all samples of pT1 and pT2 (Figure 4(a)). Scanning electron microscopy revealed superficial cells of different sizes, but prevailing ones were smaller than in normal urothelium (Figure 4(b)). They were covered with microvilli (Figure 4(b)), which are found only on poorly differentiated superficial urothelial cells [6].

Bottom Line: Terminal differentiation of urothelium is a prerequisite for blood-urine barrier formation and enables normal sensory function of the urinary bladder.In this study, urothelial differentiation of normal human urothelium and of low and high grade papillary urothelial carcinomas was correlated with the expression and localization of purinergic receptors (P2X3, and P2X5) and transient receptor potential vanilloid channels (TRPV1, and TRPV4).Our results suggest that sensory proteins P2X3 and TRPV1 are in correlation with urothelial differentiation, while P2X5 and TRPV4 have unique expression patterns.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, University Medical Centre Ljubljana, Zaloška cesta 2, 1000 Ljubljana, Slovenia.

ABSTRACT
Terminal differentiation of urothelium is a prerequisite for blood-urine barrier formation and enables normal sensory function of the urinary bladder. In this study, urothelial differentiation of normal human urothelium and of low and high grade papillary urothelial carcinomas was correlated with the expression and localization of purinergic receptors (P2X3, and P2X5) and transient receptor potential vanilloid channels (TRPV1, and TRPV4). Western blotting and immunofluorescence of uroplakins together with scanning electron microscopy of urothelial apical surface demonstrated terminal differentiation of normal urothelium, partial differentiation of low grade carcinoma, and poor differentiation of high grade carcinoma. P2X3 was expressed in normal urothelium as well as in low grade carcinoma and in both cases immunolabeling was stronger in the superficial cells. P2X3 expression decreased in high grade carcinoma. P2X5 expression was detected in normal urothelium and in high grade carcinoma, while in low grade carcinoma its expression was diminished. The expression of TRPV1 decreased in low grade and even more in high grade carcinoma when compared with normal urothelium, while TRPV4 expression was unchanged in all samples. Our results suggest that sensory proteins P2X3 and TRPV1 are in correlation with urothelial differentiation, while P2X5 and TRPV4 have unique expression patterns.

Show MeSH
Related in: MedlinePlus