Abstract
Previous reports describing Cajal‐like interstitial cells in human uterus are contradictory in terms of c‐kit immunoreactivity: either negative (but vimentin‐positive) in pregnant myometrium, or positive, presumably in the endometrium. The aim of this study was to verify the existence of human myometrial Cajal‐like interstitial cells (m‐CLIC). Six different, complementary approaches were used: 1) methylene‐blue supravital staining of tissue samples (cryosections), 2) methylene blue and Janus green B vital staining (m‐CLIC and mitochondrial markers, respectively), and 3) extracellular single‐unit electrophysiological recordings in cell cultures, 4) non‐conventional light microscopy on glutaraldehyde/osmium fixed, Epon‐embedded semi‐thin sections (less than 1μm) stained with toluidine blue (TSM), 5) transmission electron microscopy (TEM), and 6) immunofluorescence (IF). We found m‐CLIC in myometrial cryosections and in cell cultures. In vitro, m‐CLIC represented ∼7% of the total cell number. m‐CLIC had 2–3 characteristic processes which were very long (∼ 60 μm), very thin (±0.5μm) and moniliform. The dilated portions of processes usually accomodated mitochondria. In vitro, m‐CLIC exhibited spontaneous electrical activity (62.4 ± 7.22 mV field potentials, short duration: 1.197 ± 0.04ms). Moreover, m‐CLIC fulfilled the usual TEM criteria, the so‐called ‘gold’ or ‘platinum’ standards (e.g. the presence of discontinuos basal lamina, caveolae, endoplasmic reticulum, and close contacts between each other, with myocytes, nerve fibers and/or capillaries etc.). IF showed that m‐CLIC express CD117/c‐kit, sometimes associated with CD34 and with vimentin along their processes.
In conclusion, we describe myometrial Cajal‐like interstitial cells that have affinity for methylene blue and Janus green B vital dyes, fulfill (all) TEM criteria, express CD117/c‐kit and have spontaneous electric activity.
References
- 1. Thuneberg L, One hundred years of interstitial cells of Cajal. Microsc Res Tech. 1999; 47: 223–38. [DOI] [PubMed] [Google Scholar]
- 2. Faussone‐Pellegrini MS, Pantalone D, Cortesini C. Smooth muscle cells, interstitial cells of Cajal and myenteric plexus interrelationships in the human colon. Acta Anat (Basel). 1990; 139: 31–44. [DOI] [PubMed] [Google Scholar]
- 3. Rumessen JJ, Peters S, Thuneberg L. Light‐ and electron microscopical studies of interstitial cells of Cajal (ICC) and muscle cells at the submucosal border of human colon, Lab Invest. 1993; 68: 481–95. [PubMed] [Google Scholar]
- 4. Huizinga JD, Thuneberg L, Vanderwinden JM, Rumessen JJ. Interstitial cells of Cajal as targets for pharmacological intervention in gastrointestinal motor disorders. Trends Pharmacol Sci. 1997; 18: 393–403. [DOI] [PubMed] [Google Scholar]
- 5. Ward SM, Sanders KM. Physiology and pathophysiology of the interstitial cell of Cajal: From Bench to Bedside I. Functional development and plasticity of interstitial cells of Cajal networks. Am J Physiol Gastrointest Liver Physiol. 2001; 281: G602–G611. [DOI] [PubMed] [Google Scholar]
- 6. Hirst GD, Ward SM. Interstitial cells: involvement in rhythmicity and neural control of gut smooth muscle. J Physiol. 2003; 550: 337–46. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 7. Pezzone MA, Watkins SC, Alber SM, King WE, de Groat CW, Chancellor MB, Fraser MO. Identification of c‐kit‐positive cells in the mouse ureter: the interstitial cells of Cajal of the urinary tract. Am J Physiol Renal Physiol. 2003; 284: 925–9. [DOI] [PubMed] [Google Scholar]
- 8. Metzger R, Schuster T, Till H, Franke FE, Dietz HG. Cajal‐like cells in the upper urinary tract: comparative study in various species. Pediatr Surg Int. 2005; 21: 169–74. [DOI] [PubMed] [Google Scholar]
- 9. McCloskey KD, Gurney AM. Kit positive cells in the guinea‐pig bladder. J Urol. 2002; 168: 832–6. [PubMed] [Google Scholar]
- 10. Blyweert W, Aa F, Ost D, Stagnaro M, Ridder D. Interstitial cells of the bladder: the missing link BJOG. 2004; 111(s1): 57–60. [DOI] [PubMed] [Google Scholar]
- 11. Davidson RA, McCloskey KD. Morphology and localization of interstitial cells in the guinea pig bladder: structural relationships with smooth muscle and neurons. J Urol. 2005; 173: 1385–90. [DOI] [PubMed] [Google Scholar]
- 12. Sergeant GP, Hollywood MA, McCloskey KD, Thornbury KD, McHale NG. Specialized pacemaking cells in the rabbit urethra. J Physiol. 2000; 526: 359–66. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 13. Burton LD, Housley GD, Salih SG, Jaeenwood D. P2X2receptor expression by interstitial cells of Cajal in vas deferens implicated in semen emission. Auton Neurosci Basic Clin. 2000; 84: 147–61. [DOI] [PubMed] [Google Scholar]
- 14. Popescu LM, Ciontea MS, Cretoiu D, Hinescu ME, Radu E, Ionescu N, Ceausu M, Gherghiceanu M, Braga RI, Vasilescu F, Zagrean L, Ardeleanu C. Novel type of interstitial cell (Cajal‐like) in human fallopian tube. J Cell Mol Med. 2005; 9: 479–523. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 15. Harhun MI, Gordienko DV, Povstyan OV, Moss RF, Bolton TB. Function of interstitial cells of Cajal in the rabbit portal vein. Circ Res. 2004; 95: 619–26. [DOI] [PubMed] [Google Scholar]
- 16. Harhun MI, Pucovsky V, Gordienko DV, Povstyan OV, Bolton TB. Interstitial cells in the vasculature. J Cell Mol Med. 2005; 9: 232–43. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 17. McCloskey KD, Hollywood MA, Thornbury KD, Ward SM, McHale NG. Kit‐like immunopositive cells in sheap mesenteric lymphatic vessels. Cell Tissue Res. 2002; 310: 77–84. [DOI] [PubMed] [Google Scholar]
- 18. Popescu LM, Hinescu ME, Ionescu N, Ciontea MS, Cretoiu D, Ardeleanu C. Interstitial cells of Cajal in pancreas, J Cell Mol Med. 2005; 9: 169–90. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 19. Exintaris B, Klemm MF, Lang RJ. Spontaneous slow wave and contractile activity of the guinea pig prostate. J Urol. 2002; 168: 315–22. [PubMed] [Google Scholar]
- 20. van den Aa F, Roskams T, Blyweert W, de Ridder D, Interstitial cells in the human prostate: a new therapeutic target Prostate. 2003; 56: 250–5. [DOI] [PubMed] [Google Scholar]
- 21. Popescu LM, Andrei F, Hinescu ME. Snapshots of mammary gland interstitial cells: methylene blue vital staining and c‐kit immunopositivity. J Cell Mol Med. 2005; 9: 476–7. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 22. Duquette RA, Shmygol A, Vaillant C, Mobasheri A, Pope M, Burdyga T, Wray S, Vimentin‐positive, c‐kit‐negative interstitial cells in human and rat uterus: a role in pacemaking Biol Reprod. 2005; 72: 276–83. [DOI] [PubMed] [Google Scholar]
- 23. Shafik A, El‐Sibai O, Shafik I. Identification of c‐kit‐positive cells in the uterus. Int J Gynaecol Obstet. 2004; 87: 254–5. [DOI] [PubMed] [Google Scholar]
- 24. Morimoto T, Head JR, MacDonald PC, Casey ML. Thrombospondin‐1 expression in human myometrium before and during pregnancy, before and during labor, and in human myometrial cells in culture. Biol Reprod. 1998; 59: 862–70. [DOI] [PubMed] [Google Scholar]
- 25. Li CX, Liu BH, Tong WD, Zhang LY, Jiang YP. Dissociation, culture and morphologic changes of interstitial cells of Cajal in vitro. World J Gastroenterol. 2005; 11: 2838–40. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 26. Niculescu I. An atlas concerning morphological aspects of visceral nerve endings. 1st ed Bucharest : Editura Medicala; 1958. [Google Scholar]
- 27. Mora R, Bonilha VL, Marmorstein A, Scherer PE, Brown D, Lisanti MP, Rodriguez‐Boulan E. Caveolin‐2 localizes to the golgi complex but redistributes to plasma membrane, caveolae, and rafts when co‐expressed with caveolin‐1. J Biol Chem. 1999; 274: 25708–17. [DOI] [PubMed] [Google Scholar]
- 28. Cajal SR. Les nouvelles idees sur la structure du systeme nerveux chez l'homme et chez les vertebres. 2eme ed Paris : C. Reinwald & Cie, Libraires‐Editeurs, 1895. [Google Scholar]
- 29. Novikoff AB. Mitochondria in: The Cell ed. ByBrachet J., Mirsky A., Academic Press Inc., New York , 1961, 299–421. [Google Scholar]
- 30. Farrugia G. Ionic conductances in gastrointestinal smooth muscles and interstitial cells of Cajal. Annu Rev Physiol. 1999; 61: 45–84. [DOI] [PubMed] [Google Scholar]
- 31. Moore ED, Voigt T, Kobayashi YM, Isenberg G, Fay FS, Gallitelli MF, Franzini‐Armstrong C. Organization of Ca2+ release units in excitable smooth muscle of the guinea‐pig urinary bladder. Biophys J. 2004; 87: 1836–47. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 32. Erratum in: Biophys J. 2004; 87: 2914. [Google Scholar]
- 32. Wu JJ, Rothman TP, Gershon MD, Development of the interstitial cell of cajal: origin, kit dependence and nonneuronal sources of kit ligand. J Neurosci Res. 2000; 59: 384–401. [DOI] [PubMed] [Google Scholar]
- 33. Torihashi S, Ward SM, Nishikawa S, Nishi K, Kobayashi S, Sanders KM, C‐kit‐dependent development of interstitial cells and electrical activity in the murine gastrointestinal tract. Cell Tissue Res. 1995; 280: 97. [DOI] [PubMed] [Google Scholar]
- 34. Horiguchi K, Keef KD, Ward SM. Distribution of interstitial cells of Cajal in tunica muscularis of the canine rectoanal region. Am J Physiol Gastrointest Liver Physiol. 2003; 284: 756–67. [DOI] [PubMed] [Google Scholar]
- 35. Wang XY, Berezin I, Mikkelsen HB, Der T, Bercik P, Collins SM, Huizinga JD. Pathology of interstitial cells of Cajal in relation to inflammation revealed by ultrastructure but not immunohistochemistry. Am J Pathol. 2002; 160: 1529–40. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 36. Went PT, Dirnhofer S, Bundi M, Mirlacher M, Schraml P, Mangialaio S, Dimitrijevic S, Kononen J, Lugli A, Simon R, Sauter G. Prevalence of KIT expression in human tumors. J Clin Oncol. 2004; 22: 4514–22. [DOI] [PubMed] [Google Scholar]
- 37. Rushing RS, Shajahan S, Chendil D, Wilder JL, Pulliam J, Lee EY, Ueland FR, van Nagell JR, Ahmed MM, Lele SM. Uterine sarcomas express KIT protein but lack mutation(s) in exon 11 or 17 of c‐KIT. Gynecol Oncol. 2003; 91: 9–14. [DOI] [PubMed] [Google Scholar]
- 38. Sanders K., Ördög T, Ward SM. Physiology and pathophysiology of the interstitial cells of Cajal: from bench to bedside: IV. Genetic and animal models of GI motility disorders caused by loss of interstitial cells of Cajal. Am J Physiol Gastrointest Liver Physiol. 2002; 282: 747–56. [DOI] [PubMed] [Google Scholar]
- 39. Rustom A, Saffrich R, Markovic I, Walther P, Gerdes H‐H. Nanotubular highways for intercellular organelle transport. Science. 2004; 303: 1007–10. [DOI] [PubMed] [Google Scholar]
- 40. Vidulescu C, Clejan S, O'Connor KC. Vesicle traffic through intercellular bridges in DU 145 human prostate cancer cells. J Cell Mol Med. 2004; 8: 388–96. [DOI] [PMC free article] [PubMed] [Google Scholar]