Histochemistry and Cell Biology

, Volume 149, Issue 5, pp 491–501 | Cite as

Comparative immunohistochemical characterization of interstitial cells in the urinary bladder of human, guinea pig and pig

  • Clara Steiner
  • Thomas Gevaert
  • Roman Ganzer
  • Dirk De Ridder
  • Jochen NeuhausEmail author
Original Paper


Interstitial cells (ICs) are thought to play a functional role in urinary bladder. Animal models are commonly used to elucidate bladder physiology and pathophysiology. However, inter-species comparative studies on ICs are rare. We therefore analyzed ICs and their distribution in the upper lamina propria (ULP), the deeper lamina propria (DLP) and the detrusor muscular layer (DET) of human, guinea pig (GP) and pig. Paraffin slices were examined by immunohistochemistry and 3D confocal immunofluorescence of the mesenchymal intermediate filament vimentin (VIM), alpha-smooth muscle actin (αSMA), platelet-derived growth factor receptor alpha (PDGFRα) and transient receptor potential cation channel A1 (TRPA1). Image stacks were processed for analysis using Huygens software; quantitative analysis was performed with Fiji macros. ICs were identified by immunoreactivity for VIM (excluding blood vessels). In all species ≥ 75% of ULP ICs were VIM+/PDGFRα+ and ≥ 90% were VIM+/TRPA1+. In human and pig ≥ 74% of ULP ICs were VIM+/αSMA+, while in GP the percentage differed significantly with only 37% VIM+/αSMA+ ICs. Additionally, over 90% of αSMA+ ICs were also TRPA1+ and PDGFRα+ in human, GP and pig. In all three species, TRPA1+ and PDGFRα+ ICs point to an active role for these cells in bladder physiology, regarding afferent signaling processes and signal modification. We hypothesize that decline in αSMA-positivity in GP reflects adaptation of bladder histology to smaller bladder size. In our experiments, pig bladder proved to be highly comparable to human urinary bladder and seems to provide safer interpretation of experimental findings than GP.


3D confocal laser scanning microscopy Quantitative analysis PDGFR-alpha TRPA1 Alpha-smooth muscle actin Vimentin 



The authors thank Mrs. Annett Weimann, Mrs. Mandy Bernd-Paetz (Leipzig) and Mrs. Nathalie Volders (Leuven) for the excellent technical assistance. We thank Dr. Thomas Pannicke (Paul-Flechsig Intitute of Brain Research, University of Leipzig) for providing guinea-pig tissue.


This study was in part funded by the Dr. Siegfried Krüger Stiftung Leipzig.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval/standards

All procedures were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. Human studies were approved by the Ethical Committee of the University of Leipzig (UKL8/2004) and informed consent was obtained from all patients included in this study.

Supplementary material

418_2018_1655_MOESM1_ESM.tif (3.5 mb)
Fig. S1. IF in GP supporting TRPA1-antibody validation data using a second antibody against TRPA1; upper panels (a - c) show main antibody from alomone (ACC-037), lower panels (d - f) show second antibody from Santa Cruz (sc-32353) in bladder and gut. Asterisks show TRPA1-IR in enteric ganglia. Bladder stains with blocking peptides show no IR for TRPA1. Channels: TRPA1 in red, VIM in green; nuclei in blue (DAPI); U = Urothelium readily discernible by characteristic arrangement of nuclei. Scale bar indicates 20 µm. (TIF 3615 KB)
418_2018_1655_MOESM2_ESM.tif (480 kb)
Fig. S2. Validation of alomone anti-TRPA1 antibody in WB; banding was closely resembling the WB reported in the datasheet. First four bands show housekeeping genes β-actin and GAPDH. Each antibody was used in two different GPs. (TIF 479 KB)
418_2018_1655_MOESM3_ESM.tif (415 kb)
Fig. S3. Parts-of-a-whole diagrams show analysis of αSMA/PDGFRα and αSMA/TRPA1 in human, GP and pig. Amount of PDGFRα- and TRPA1- ICs with positive αSMA-IR is <10% in all species. GP shows less αSMA+ cells than human and pig. (TIF 414 KB)
418_2018_1655_MOESM4_ESM.avi (684 kb)
Video S4. 360° Projection of a 45 optical slices confocal stack of human ICs (see figure 2a); Channels: PDGFRα in red, Vimentin in green and DAPI in blue. (AVI 684 KB)
418_2018_1655_MOESM5_ESM.avi (918 kb)
Video S5. 360° Projection of a 45 optical slices confocal stack of GP ICs (see figure 2b); Channels: PDGFRα in red, Vimentin in green and DAPI in blue. (AVI 917 KB)
418_2018_1655_MOESM6_ESM.avi (907 kb)
Video S6. 360° Projection of a 45 optical slices confocal stack of pig ICs (see figure 2c); Channels: PDGFRα in red, Vimentin in green and DAPI in blue. (AVI 906 KB)


  1. Andersson K-E, McCloskey KD (2014) Lamina propria: the functional center of the bladder? Neurourol Urodyn 33(1):9–16. CrossRefPubMedGoogle Scholar
  2. Andersson K-E, Gratzke C, Hedlund P (2010) The role of the transient receptor potential (TRP) superfamily of cation-selective channels in the management of the overactive bladder. BJU Int 106(8):1114–1127. CrossRefPubMedGoogle Scholar
  3. Baker SA, Hennig GW, Salter AK, Kurahashi M, Ward SM, Sanders KM (2013) Distribution and Ca(2+) signalling of fibroblast-like (PDGFR(+)) cells in the murine gastric fundus. J Physiol (Lond) 591(24):6193–6208. CrossRefGoogle Scholar
  4. Bastiani M, Parton RG (2010) Caveolae at a glance. J Cell Sci 123(Pt 22):3831–3836. CrossRefPubMedGoogle Scholar
  5. Blyweert W, Aa F, OST D, Stagnaro M, Ridder D (2004) Interstitial cells of the bladder: the missing link? BJOG 111(s1):57–60. CrossRefPubMedGoogle Scholar
  6. Cunningham RMJ, Larkin P, McCloskey KD (2011) Ultrastructural properties of interstitial cells of Cajal in the Guinea pig bladder. J Urol 185(3):1123–1131. CrossRefPubMedGoogle Scholar
  7. Davidson RA, McCloskey KD (2005) Morphology and localization of interstitial cells in the guinea pig bladder: structural relationships with smooth muscle and neurons. J Urol 173(4):1385–1390. CrossRefPubMedGoogle Scholar
  8. Demoulin J-B, Essaghir A (2014) PDGF receptor signaling networks in normal and cancer cells. Cytokine Growth Factor Rev 25(3):273–283. CrossRefPubMedGoogle Scholar
  9. Du S, Araki I, Yoshiyama M, Nomura T, Takeda M (2007) Transient receptor potential channel A1 involved in sensory transduction of rat urinary bladder through C-fiber pathway. Urology 70(4):826–831. CrossRefPubMedGoogle Scholar
  10. Franken J, Uvin P, Ridder D de, Voets T (2014) TRP channels in lower urinary tract dysfunction. Br J Pharmacol 171(10):2537–2551. CrossRefPubMedPubMedCentralGoogle Scholar
  11. Fry CH, Sui G-P, Kanai AJ, Wu C (2007) The function of suburothelial myofibroblasts in the bladder. Neurourol Urodyn 26(6 Suppl):914–919. CrossRefPubMedGoogle Scholar
  12. Gevaert T, Vos R de, Everaerts W, Libbrecht L, Van Der Aa F, van den Oord J et al (2011) Characterization of upper lamina propria interstitial cells in bladders from patients with neurogenic detrusor overactivity and bladder pain syndrome. J Cell Mol Med 15(12):2586–2593. CrossRefPubMedPubMedCentralGoogle Scholar
  13. Gevaert T, Vos R de, Van Der Aa F, Joniau S, van den Oord J, Roskams T et al (2012) Identification of telocytes in the upper lamina propria of the human urinary tract. J Cell Mol Med 16(9):2085–2093. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Gevaert T, Hutchings G, Everaerts W, Prenen H, Roskams T, Nilius B et al (2014a) Administration of imatinib mesylate in rats impairs the neonatal development of intramuscular interstitial cells in bladder and results in altered contractile properties. Neurourol Urodyn 33(4):461–468. CrossRefPubMedGoogle Scholar
  15. Gevaert T, Vanstreels E, Daelemans D, Franken J, Van Der Aa F, Roskams T et al (2014b) Identification of different phenotypes of interstitial cells in the upper and deep lamina propria of the human bladder dome. J Urol 192(5):1555–1563. CrossRefPubMedGoogle Scholar
  16. Gevaert T, Moles Lopez X, Sagaert X, Libbrecht L, Roskams T, Rorive S et al (2015) Morphometric and quantitative immunohistochemical analysis of disease-related changes in the upper (suburothelial) lamina propria of the human bladder dome. PLoS One 10(5):e0127020. CrossRefPubMedPubMedCentralGoogle Scholar
  17. Gevaert T, Neuhaus J, Vanstreels E, Daelemans D, Everaerts W, Van Der Aa F et al (2017a) Comparative study of the organisation and phenotypes of bladder interstitial cells in human, mouse and rat. Cell Tissue Res 370(3):403–416. CrossRefPubMedGoogle Scholar
  18. Gevaert T, Ridder D de, Vanstreels E, Daelemans D, Everaerts W, Van Der Aa F et al (2017b) The stem cell growth factor receptor KIT is not expressed on interstitial cells in bladder. J Cell Mol Med 21(6):1206–1216. CrossRefPubMedGoogle Scholar
  19. Gratzke C, Weinhold P, Reich O, Seitz M, Schlenker B, Stief CG et al (2010) Transient receptor potential A1 and cannabinoid receptor activity in human normal and hyperplastic prostate. Relation to nerves and interstitial cells. Eur Urol 57(5):902–910. CrossRefPubMedGoogle Scholar
  20. Grol S, Essers PBM, van Koeveringe GA, Martinez-Martinez P, Vente J de, Gillespie JI (2009) M(3) muscarinic receptor expression on suburothelial interstitial cells. BJU Int 104(3):398–405. CrossRefPubMedGoogle Scholar
  21. Grol S, Nile CJ, Martinez-Martinez P, van Koeveringe G, Wachter S de, Vente J de et al (2011) M3 muscarinic receptor-like immunoreactivity in sham operated and obstructed guinea pig bladders. J Urol 185(5):1959–1966. CrossRefPubMedGoogle Scholar
  22. Hashitani H, Yanai Y, Suzuki H (2004) Role of interstitial cells and gap junctions in the transmission of spontaneous Ca2 + signals in detrusor smooth muscles of the guinea-pig urinary bladder. J Physiol (Lond) 559(Pt 2):567–581. CrossRefGoogle Scholar
  23. Heldin CH, Lennartsson J, Westermark B (2018) Involvement of platelet-derived growth factor ligands and receptors in tumorigenesis. J Intern Med 283(1):16–44. CrossRefPubMedGoogle Scholar
  24. Johnston L, Woolsey S, Cunningham RMJ, O’Kane H, Duggan B, Keane P et al (2010) Morphological expression of KIT positive interstitial cells of Cajal in human bladder. J Urol 184(1):370–377. CrossRefPubMedPubMedCentralGoogle Scholar
  25. Koh BH, Roy R, Hollywood MA, Thornbury KD, McHale NG, Sergeant GP et al (2012) Platelet-derived growth factor receptor-α cells in mouse urinary bladder: a new class of interstitial cells. J Cell Mol Med 16(4):691–700. CrossRefPubMedPubMedCentralGoogle Scholar
  26. Koh SD, Lee H, Ward SM, Sanders KM (2018) The mystery of the interstitial cells in the urinary bladder. Annu Rev Pharmacol Toxicol 58:603–623. CrossRefPubMedGoogle Scholar
  27. Lee H, Koh BH, Peri LE, Sanders KM, Koh SD (2014) Purinergic inhibitory regulation of murine detrusor muscles mediated by PDGFRalpha + interstitial cells. J Physiol (Lond) 592(6):1283–1293. CrossRefGoogle Scholar
  28. Liu F, Takahashi N, Yamaguchi O (2009) Expression of P2 × 3 purinoceptors in suburothelial myofibroblasts of the normal human urinary bladder. Int J Urol 16(6):570–575. CrossRefPubMedGoogle Scholar
  29. McCloskey KD, Gurney AM (2002) Kit positive cells in the guinea pig bladder. J Urol 168(2):832–836CrossRefPubMedGoogle Scholar
  30. McHale NG, Hollywood MA, Sergeant GP, Shafei M, Thornbury KT, Ward SM (2006) Organization and function of ICC in the urinary tract. J Physiol (Lond) 576(Pt 3):689–694. CrossRefGoogle Scholar
  31. Merryman WD, Youn I, Lukoff HD, Krueger PM, Guilak F, Hopkins RA et al (2006) Correlation between heart valve interstitial cell stiffness and transvalvular pressure. Implications for collagen biosynthesis. Am J Physiol Heart Circ Physiol 290(1):H224-31. CrossRefPubMedGoogle Scholar
  32. Monaghan KP, Johnston L, McCloskey KD (2012) Identification of PDGFRα positive populations of interstitial cells in human and guinea pig bladders. J Urol 188(2):639–647. CrossRefPubMedGoogle Scholar
  33. Nagata K, Duggan A, Kumar G, García-Añoveros J (2005) Nociceptor and hair cell transducer properties of TRPA1, a channel for pain and hearing. J Neurosci 25(16):4052–4061. CrossRefPubMedGoogle Scholar
  34. Neuhaus J, Heinrich M, Schlichting N, Oberbach A, Fitzl G, Schwalenberg T et al (2007) Struktur und Funktion der suburothelialen Myofibroblasten in der humanen Harnblase unter normalen und pathologischen Bedingungen (Structure and function of suburothelial myofibroblasts in the human urinary bladder under normal and pathological conditions). Urologe A 46(9):1197–1202. CrossRefPubMedGoogle Scholar
  35. Neuhaus J, Schröppel B, Dass M, Zimmermann H, Wolburg H, Fallier-Becker P et al (2018) 3D-electron microscopic characterization of interstitial cells in the human bladder upper lamina propria. Neurourol Urodyn 37:89–98. CrossRefPubMedGoogle Scholar
  36. Oyama S, Dogishi K, Kodera M, Kakae M, Nagayasu K, Shirakawa H et al (2017) Pathophysiological role of transient receptor potential ankyrin 1 in a mouse long-lasting cystitis model induced by an intravesical injection of hydrogen peroxide. Front Physiol 8:877. CrossRefPubMedPubMedCentralGoogle Scholar
  37. Piaseczna Piotrowska A, Rolle U, Solari V, Puri P (2004) Interstitial cells of Cajal in the human normal urinary bladder and in the bladder of patients with megacystis-microcolon intestinal hypoperistalsis syndrome. BJU Int 94(1):143–146. CrossRefPubMedGoogle Scholar
  38. Rusu MC, Folescu R, Mănoiu VS, Didilescu AC (2014) Suburothelial interstitial cells. Cells Tissues Organs 199(1):59–72. CrossRefPubMedGoogle Scholar
  39. Sadananda P, Chess-Williams R, Burcher E (2008) Contractile properties of the pig bladder mucosa in response to neurokinin A: a role for myofibroblasts? Br J Pharmacol 153(7):1465–1473. CrossRefPubMedPubMedCentralGoogle Scholar
  40. Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T et al (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9(7):676–682. CrossRefPubMedGoogle Scholar
  41. Shafik A, El-Sibai O, Shafik AA, Shafik I (2004) Identification of interstitial cells of Cajal in human urinary bladder: concept of vesical pacemaker. Urology 64(4):809–813. CrossRefPubMedGoogle Scholar
  42. Story GM, Peier AM, Reeve AJ, Eid SR, Mosbacher J, Hricik TR et al (2003) ANKTM1, a TRP-like channel expressed in nociceptive neurons, is activated by cold temperatures. Cell 112(6):819–829CrossRefPubMedGoogle Scholar
  43. Streng T, Axelsson HE, Hedlund P, Andersson DA, Jordt S-E, Bevan S et al (2008) Distribution and function of the hydrogen sulfide-sensitive TRPA1 ion channel in rat urinary bladder. Eur Urol 53(2):391–399. CrossRefPubMedGoogle Scholar
  44. Sui GP, Wu C, Fry CH (2004) Electrical characteristics of suburothelial cells isolated from the human bladder. J Urol 171(2 Pt 1):938–943. CrossRefPubMedGoogle Scholar
  45. Vannucchi M-G, Traini C, Guasti D, Del Popolo G, Faussone-Pellegrini M-S (2014) Telocytes subtypes in human urinary bladder. J Cell Mol Med 18(10):2000–2008. CrossRefPubMedPubMedCentralGoogle Scholar
  46. Weinhold P, Hennenberg M, Strittmatter F, Stief CG, Gratzke C, Hedlund P (2017) Transient receptor potential a1 (TRPA1) agonists inhibit contractions of the isolated human ureter. Neurourol Urodyn. PubMedCrossRefGoogle Scholar
  47. Wiseman OJ, Fowler CJ, Landon DN (2003) The role of the human bladder lamina propria myofibroblast. BJU Int 91(1):89–93CrossRefPubMedGoogle Scholar
  48. Yu W, Zeidel ML, Hill WG (2012) Cellular expression profile for interstitial cells of cajal in bladder—a cell often misidentified as myocyte or myofibroblast. PloS one 7(11):e48897. CrossRefPubMedPubMedCentralGoogle Scholar
  49. Zheng Y, Zhu T, Lin M, Wu D, Wang X (2012) Telocytes in the urinary system. J Transl Med 10:188. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Urology, Research LaboratoriesUniversity of LeipzigLeipzigGermany
  2. 2.Organ Systems, Laboratory of Experimental UrologyKU LeuvenLeuvenBelgium
  3. 3.Department of UrologyUZ LeuvenLeuvenBelgium
  4. 4.Department of UrologyUniversity Hospital Leipzig AöRLeipzigGermany
  5. 5.Department of UrologyAsklepios Hospital Bad TölzBad TölzGermany

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