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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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The expression pattern of uroplakins, P2X3, P2X5, TRPV1, and TRPV4 in normal human urothelium and in low and high grade papillary urothelial carcinomas as determined by Western blotting. In the protein samples of normal urothelium, UPIIIa, P2X3, P2X5, TRPV1, and TRPV4 are expressed. In low grade carcinoma, there is no expression of uroplakins. P2X3 and TRPV4 are expressed as in normal urothelium, while P2X5 is greatly diminished. TRPV1 expression is decreased in comparison to normal urothelium. In the protein samples of high grade carcinoma with lamina propria invasion (pT1) and those with muscularis propria invasion (pT2), the expression patterns were similar and therefore three examples of each are presented here. The expression of uroplakins is negative and expressions of P2X3 and TRPV1 are substantially decreased compared to normal urothelium. The expressions of P2X5 and TRPV4 are the same as in normal urothelium. Western blots were done in duplicate. Molecular weights are shown in kilodaltons (kDa).
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fig1: The expression pattern of uroplakins, P2X3, P2X5, TRPV1, and TRPV4 in normal human urothelium and in low and high grade papillary urothelial carcinomas as determined by Western blotting. In the protein samples of normal urothelium, UPIIIa, P2X3, P2X5, TRPV1, and TRPV4 are expressed. In low grade carcinoma, there is no expression of uroplakins. P2X3 and TRPV4 are expressed as in normal urothelium, while P2X5 is greatly diminished. TRPV1 expression is decreased in comparison to normal urothelium. In the protein samples of high grade carcinoma with lamina propria invasion (pT1) and those with muscularis propria invasion (pT2), the expression patterns were similar and therefore three examples of each are presented here. The expression of uroplakins is negative and expressions of P2X3 and TRPV1 are substantially decreased compared to normal urothelium. The expressions of P2X5 and TRPV4 are the same as in normal urothelium. Western blots were done in duplicate. Molecular weights are shown in kilodaltons (kDa).

Mentions: Uroplakins, differentiation dependent and urothelium-specific transmembrane proteins, were detected by polyclonal anti-uroplakin antibody, which reacts strongly with UPIIIa (47 kDa). The expression of UPIIIa in all samples of normal human urothelium was positive (Figure 1). To localize uroplakins within urothelium we used immunofluorescence. Normal urothelium showed strong uroplakin labelling of the superficial cells (Figure 2(a)). Scanning electron microscopy revealed large, polygonal cells covering urothelial surface (Figure 2(b)). They had scalloped appearance with microridges, demonstrating the presence of urothelial plaques. All features indicated above provided evidence that umbrella cells formed the superficial cell layer of normal urothelium, and they were therefore considered to be terminally differentiated. Confirming this, we further analysed the expression and distribution of four nonselective ion channels.


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)

The expression pattern of uroplakins, P2X3, P2X5, TRPV1, and TRPV4 in normal human urothelium and in low and high grade papillary urothelial carcinomas as determined by Western blotting. In the protein samples of normal urothelium, UPIIIa, P2X3, P2X5, TRPV1, and TRPV4 are expressed. In low grade carcinoma, there is no expression of uroplakins. P2X3 and TRPV4 are expressed as in normal urothelium, while P2X5 is greatly diminished. TRPV1 expression is decreased in comparison to normal urothelium. In the protein samples of high grade carcinoma with lamina propria invasion (pT1) and those with muscularis propria invasion (pT2), the expression patterns were similar and therefore three examples of each are presented here. The expression of uroplakins is negative and expressions of P2X3 and TRPV1 are substantially decreased compared to normal urothelium. The expressions of P2X5 and TRPV4 are the same as in normal urothelium. Western blots were done in duplicate. Molecular weights are shown in kilodaltons (kDa).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4020497&req=5

fig1: The expression pattern of uroplakins, P2X3, P2X5, TRPV1, and TRPV4 in normal human urothelium and in low and high grade papillary urothelial carcinomas as determined by Western blotting. In the protein samples of normal urothelium, UPIIIa, P2X3, P2X5, TRPV1, and TRPV4 are expressed. In low grade carcinoma, there is no expression of uroplakins. P2X3 and TRPV4 are expressed as in normal urothelium, while P2X5 is greatly diminished. TRPV1 expression is decreased in comparison to normal urothelium. In the protein samples of high grade carcinoma with lamina propria invasion (pT1) and those with muscularis propria invasion (pT2), the expression patterns were similar and therefore three examples of each are presented here. The expression of uroplakins is negative and expressions of P2X3 and TRPV1 are substantially decreased compared to normal urothelium. The expressions of P2X5 and TRPV4 are the same as in normal urothelium. Western blots were done in duplicate. Molecular weights are shown in kilodaltons (kDa).
Mentions: Uroplakins, differentiation dependent and urothelium-specific transmembrane proteins, were detected by polyclonal anti-uroplakin antibody, which reacts strongly with UPIIIa (47 kDa). The expression of UPIIIa in all samples of normal human urothelium was positive (Figure 1). To localize uroplakins within urothelium we used immunofluorescence. Normal urothelium showed strong uroplakin labelling of the superficial cells (Figure 2(a)). Scanning electron microscopy revealed large, polygonal cells covering urothelial surface (Figure 2(b)). They had scalloped appearance with microridges, demonstrating the presence of urothelial plaques. All features indicated above provided evidence that umbrella cells formed the superficial cell layer of normal urothelium, and they were therefore considered to be terminally differentiated. Confirming this, we further analysed the expression and distribution of four nonselective ion channels.

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