Volume 53, Issue 2, Pages 391-400 (February 2008) Distribution and Function of the Hydrogen Sulfide–Sensitive TRPA1 Ion Channel in Rat Urinary Bladder  Tomi Streng, Helena E. Axelsson, Petter Hedlund, David A. Andersson, Sven-Eric Jordt, Stuart Bevan, Karl-Erik Andersson, Edward D. Högestätt, Peter M. Zygmunt  European Urology  Volume 53, Issue 2, Pages 391-400 (February 2008) DOI: 10.1016/j.eururo.2007.10.024 Copyright © 2007 Terms and Conditions

Fig. 1 Immunohistochemical localisation of TRPA1 in the bladder wall. TRPA1-IR nerve fibres colocalise with TRPV1 (A and B), CGRP (C and D), and substance P (E and F). The TRPV1 primary antibody was obtained from Santa Cruz Biotechnologies. Scale bars represent 25μm in A and B, and 100μm in all other images. European Urology 2008 53, 391-400DOI: (10.1016/j.eururo.2007.10.024) Copyright © 2007 Terms and Conditions

Fig. 2 Immunohistochemical localisation of TRPA1 in the urothelium. TRPA1-IR nerve fibres are present in the suburothelial region and within the urothelium. Urothelial cells also display TRPA1 immunoreactivity. TRPV1 immunoreactivity is found in nerve fibres in the suburothelial region and within the urothelium as well as in urothelial cells with the strongest staining of the superficial cell layer. The TRPV1 primary antibody was obtained from Calbiochem. Insets show control experiments with secondary antibodies used for TRPA1 (Alexa 488 chicken antirabbit) and TRPV1 (Alexa 488 goat antirabbit) immunohistochemistry. Scale bars represent 100μm in all images. L, lumen; su, suburothelium; u, urothelium. European Urology 2008 53, 391-400DOI: (10.1016/j.eururo.2007.10.024) Copyright © 2007 Terms and Conditions

Fig. 3 Cystometrograms from three rats showing bladder pressure (upper panel) and voided volume (lower panel). (A) Effect of intravesical infusion of allyl isothiocyanate at 10μmol/l and 100μmol/l. (B) Effect of intravesical infusion of cinnamaldehyde at 500μmol/l. (C) Effect of intravesical infusion of NaHS at 1mmol/l. In B and C, the bladder was pretreated with protamine sulfate to increase drug penetration. European Urology 2008 53, 391-400DOI: (10.1016/j.eururo.2007.10.024) Copyright © 2007 Terms and Conditions

Fig. 4 Time course of NaHS (10mmol/l) evoked increases in [Ca2+]i in CHO cells expressing mouse TRPA1 (mTRPA1) or human TRPA1 (hTRPA1). (A) NaHS (50mmol/l) produced no [Ca2+]i responses in untransfected control cells (CHO). Traces shown are the mean response from triplicate wells. (B) Concentration-response curve for NaHS in mTRPA1 CHO cells (five separate experiments). European Urology 2008 53, 391-400DOI: (10.1016/j.eururo.2007.10.024) Copyright © 2007 Terms and Conditions