Pflügers Archiv - European Journal of Physiology

, Volume 469, Issue 9, pp 1189–1202 | Cite as

Properties of synchronous spontaneous Ca2+ transients in the mural cells of rat rectal arterioles

  • Retsu MitsuiEmail author
  • Hikaru Hashitani
Signaling and cell physiology
Part of the following topical collections:
  1. Signaling and cell physiology


Synchrony of spontaneous Ca2+ transients among venular mural cells (smooth muscle cells and pericytes) in visceral organs relies on the intercellular spread of L-type voltage-dependent Ca2+ channel (LVDCC)-dependent depolarisations. However, the mechanisms underlying the synchrony of spontaneous Ca2+ transients between arteriolar mural cells are less understood. The spontaneous intracellular Ca2+ dynamics of arteriolar mural cells in the rat rectal submucosa were visualised by Cal-520 Ca2+ imaging to analyse their synchrony. The mural cells in fine arterioles that had a rounded cell body with several extended processes developed spontaneous ‘synchronous’ Ca2+ transients arising from Ca2+ released from sarcoendoplasmic reticulum Ca2+ stores. Gap junction blockers (3 μM carbenoxolone, 10 μM 18β-glycyrrhetinic acid), a Ca2+-activated Cl channel (CaCC) blocker (100 μM 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid) or lowering extracellular Cl concentration (from 134.4 to 12.4 mM) disrupted the synchrony of Ca2+ transients between arteriolar mural cells. Blockers of T-type voltage-dependent Ca2+ channels (TVDCCs, 1 μM mibefradil or ML218) or LVDCCs (1 μM nifedipine) reduced the Ca2+ transient frequency or their area under curve (AUC), respectively. However, neither TVDCC nor LVDCC blockers disrupted the synchrony of Ca2+ transients among arteriolar mural cells. This is in contrast with rectal venules in which nifedipine disrupted the synchrony of spontaneous Ca2+ transients. Thus, spontaneous transient depolarisations arising from the opening of CaCCs may effectively spread to neighbouring arteriolar mural cells via gap junctions to maintain the Ca2+ transient synchrony. Activation of TVDCCs appears to accelerate spontaneous Ca2+ transients, while LVDCCs predominantly contribute to the duration of Ca2+ transients.


Blood vessel Microvasculature Smooth muscles Ca2+ signalling Intestine 



α-Smooth muscle actin


Area under curve


Ca2+-activated Cl channel


L-type voltage-dependent Ca2+ channel




NG2 chondroitin sulphate proteoglycan


Physiological salt solution


T-type voltage-dependent Ca2+ channel



The authors wish to thank Dr. Richard Lang (Monash University) for critical reading of the manuscript. The present study was partly supported by Grant-in-Aid for Young Scientists (B) (No. 26860521 and No. 16K19361) from Japan Society for Promotion of the Science (JSPS) to R.M., Grant-in-Aid for Challenging Exploratory Research (No. 26670705) from JSPS to H.H. and Grant-in-Aid from The Hori Sciences and Arts Foundation to R.M.

Compliance with ethical standards


The experimental protocols in the present study were approved by the animal experimentation ethics committee at Nagoya City University Graduate School of Medical Sciences.

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Aickin CC, Brading AF (1990) The effect of loop diuretics on Cl transport in smooth muscle of the guinea-pig vas deferens and taenia from the caecum. J Physiol 421:33–53CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Best L, Bolton TB (1986) Depolarisation of guinea-pig visceral smooth muscle causes hydrolysis of inositol phospholipids. Naunyn Schmiedeberg’s Arch Pharmacol 333:78–82CrossRefGoogle Scholar
  3. 3.
    Boedtkjer DM, Matchkov VV, Boedtkjer E, Nilsson H, Aalkjaer C (2008) Vasomotion has chloride-dependency in rat mesenteric small arteries. Pflugers Arch 457:389–404CrossRefPubMedGoogle Scholar
  4. 4.
    Boley SJ, Agrawal GP, Warren AR, Veith FJ, Levowitz BS, Treiber W, Dougherty J, Schwartz SS, Gliedman ML (1969) Pathophysiologic effects of bowel distention on intestinal blood flow. Am J Surg 117:228–234CrossRefPubMedGoogle Scholar
  5. 5.
    Borysova L, Wray S, Eisner DA, Burdyga T (2013) How calcium signals in myocytes and pericytes are integrated across in situ microvascular networks and control microvascular tone. Cell Calcium 54:163–174CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Bulley S, Jaggar JH (2014) Cl channels in smooth muscle cells. Pflugers Arch 466:861–872CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Chen RY, Li DS, Guth PH (1992) Role of calcitonin gene-related peptide in capsaicin-induced gastric submucosal arteriolar dilation. Am J Phys 262:H1350–H1355Google Scholar
  8. 8.
    Dora KA, Gallagher NT, McNeish A, Garland CJ (2008) Modulation of endothelial cell KCa3.1 channels during endothelium-derived hyperpolarizing factor signaling in mesenteric resistance arteries. Circ Res 102:1247–1255CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Edlich RF, Borner JW, Kuphal J, Wangensteen OH (1970) Gastric blood flow. I. Its distribution during gastric distention. Am J Surg 120:35–37CrossRefPubMedGoogle Scholar
  10. 10.
    Evans RJ, Surprenant A (1992) Vasoconstriction of guinea-pig submucosal arterioles following sympathetic nerve stimulation is mediated by the release of ATP. Br J Pharmacol 106:242–249CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Fujiwara T, Uehara Y (1984) The cytoarchitecture of the wall and the innervation pattern of the microvessels in the rat mammary gland: a scanning electron microscopic observation. Am J Anat 170:39–54CrossRefPubMedGoogle Scholar
  12. 12.
    Guth PH, Smith E (1975) Neural control of gastric mucosal blood flow in the rat. Gastroenterology 69:935–940PubMedGoogle Scholar
  13. 13.
    Haddock RE, Hill CE (2002) Differential activation of ion channels by inositol 1,4,5-trisphosphate (IP3)- and ryanodine-sensitive calcium stores in rat basilar artery vasomotion. J Physiol 545:615–627CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Hashitani H, Lang RJ (2016) Spontaneous activity in the microvasculature of visceral organs: role of pericytes and voltage-dependent Ca2+ channels. J Physiol 594:555–565CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Hashitani H, Takano H, Fujita K, Mitsui R, Suzuki H (2011) Functional properties of suburothelial microvessels in the rat bladder. J Urol 185:2382–2391CrossRefPubMedGoogle Scholar
  16. 16.
    Hashitani H, Mitsui R, Shimizu Y, Higashi R, Nakamura K (2012) Functional and morphological properties of pericytes in suburothelial venules of the mouse bladder. Br J Pharmacol 167:1723–1736CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Hashitani H, Mitsui R, Masaki S, van Helden DF (2015) Pacemaker role of pericytes in generating synchronised spontaneous Ca2+ transients in the myenteric microvasculature of the guinea-pig gastric antrum. Cell Calcium 58:442–456CrossRefPubMedGoogle Scholar
  18. 18.
    Hashitani H, Nguyen M, Noda H, Mitsui R, Higashi R, Ohta K, Nakamura K, Lang RJ (2017) Interstitial cell modulation of pyeloureteric peristalsis in the mouse renal pelvis using FIBSEM tomography and calcium indicators. Pflugers Archiv. doi: 10.1007/s00424-016-1930-6 Google Scholar
  19. 19.
    Hirano Y, Fozzard HA, January CT (1989) Characteristics of L- and T-type Ca2+ currents in canine cardiac Purkinje cells. Am J Phys 256:H1478–H1492Google Scholar
  20. 20.
    Hirst GD (1977) Neuromuscular transmission in arterioles of guinea-pig submucosa. J Physiol 273:263–275CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Kito Y, Suzuki H (2003) Properties of pacemaker potentials recorded from myenteric interstitial cells of Cajal distributed in the mouse small intestine. J Physiol 553:803–818CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Kito Y, Mitsui R, Ward SM, Sanders KM (2015) Characterization of slow waves generated by myenteric interstitial cells of Cajal of the rabbit small intestine. Am J Physiol Gastrointest Liver Physiol 308:G378–G388CrossRefPubMedGoogle Scholar
  23. 23.
    Krohg-Sørensen K, Lunde OC (1993) Perfusion of the human distal colon and rectum evaluated with endoscopic laser Doppler flowmetry. Methodologic aspects. Scand J Gastroenterol 28:104–108CrossRefPubMedGoogle Scholar
  24. 24.
    Lee S, Roizes S, von der Weid PY (2014) Distinct roles of L- and T-type voltage-dependent Ca2+ channels in regulation of lymphatic vessel contractile activity. J Physiol 592:5409–5427CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Little TL, Xia J, Duling BR (1995) Dye tracers define differential endothelial and smooth muscle coupling patterns within the arteriolar wall. Circ Res 76:498–504CrossRefPubMedGoogle Scholar
  26. 26.
    Mitsui R, Hashitani H (2013) Immunohistochemical characteristics of suburothelial microvasculature in the mouse bladder. Histochem Cell Biol 140:189–200CrossRefPubMedGoogle Scholar
  27. 27.
    Mitsui R, Hashitani H (2015) Functional properties of submucosal venules in the rat stomach. Pflugers Archiv 467:1327–1342CrossRefPubMedGoogle Scholar
  28. 28.
    Mitsui R, Hashitani H (2016) Mechanisms underlying spontaneous constrictions of postcapillary venules in the rat stomach. Pflugers Archiv 468:279–291CrossRefPubMedGoogle Scholar
  29. 29.
    Mitsui R, Miyamoto S, Takano H, Hashitani H (2013) Properties of submucosal venules in the rat distal colon. Br J Pharmacol 170:968–977CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Morgan KG (1983) Comparison of membrane electrical activity of cat gastric submucosal arterioles and venules. J Physiol 345:135–147CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Murfee WL, Skalak TC, Peirce SM (2005) Differential arterial/venous expression of NG2 proteoglycan in perivascular cells along microvessels: identifying a venule-specific phenotype. Microcirculation 12:151–160CrossRefPubMedGoogle Scholar
  32. 32.
    Neild TO, Shen KZ, Surprenant A (1990) Vasodilatation of arterioles by acetylcholine released from single neurones in the guinea-pig submucosal plexus. J Physiol 420:247–265CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Park F, Mattson DL, Roberts LA, Cowley AW Jr (1997) Evidence for the presence of smooth muscle alpha-actin within pericytes of the renal medulla. Am J Phys 273:R1742–R1748Google Scholar
  34. 34.
    Shimizu Y, Mochizuki S, Mitsui R, Hashitani H (2014) Neurohumoral regulation of spontaneous constrictions in suburothelial venules of the rat urinary bladder. Vasc Pharmacol 60:84–94CrossRefGoogle Scholar
  35. 35.
    Sims DE (1986) The pericyte—a review. Tissue Cell 18:153–174CrossRefPubMedGoogle Scholar
  36. 36.
    Skinner SA, O’Brien PE (1996) The microvascular structure of the normal colon in rats and humans. J Surg Res 61:482–490CrossRefPubMedGoogle Scholar
  37. 37.
    Uehara Y, Fujiwara T, Kaidoh T (1990) Morphology of vascular smooth muscle fibers and pericytes: scanning electron microscopic studies. In: Motta PM (ed) Ultrastructure of smooth muscle. Kluwer Academic Publishers, Dordrecht, pp 237–251CrossRefGoogle Scholar
  38. 38.
    Vanner S (1994) Corelease of neuropeptides from capsaicin-sensitive afferents dilates submucosal arterioles in guinea pig ileum. Am J Phys 267:G650–G655Google Scholar
  39. 39.
    Vanner S, Jiang MM, Surprenant A (1993) Mucosal stimulation evokes vasodilation in submucosal arterioles by neuronal and nonneuronal mechanisms. Am J Phys 264:G202–G212Google Scholar
  40. 40.
    Yamamoto Y, Klemm MF, Edwards FR, Suzuki H (2001) Intercellular electrical communication among smooth muscle and endothelial cells in guinea-pig mesenteric arterioles. J Physiol 535:181–195CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Zhang Q, Cao C, Zhang Z, Wier WG, Edwards A, Pallone TL (2008) Membrane current oscillations in descending vasa recta pericytes. Am J Physiol Renal Physiol 294:F656–F666CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Zhang Z, Payne K, Pallone TL (2016) Descending vasa recta endothelial membrane potential response requires pericyte communication. PLoS One 11:e0154948CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  1. 1.Department of Cell PhysiologyNagoya City University Graduate School of Medical SciencesNagoyaJapan

Personalised recommendations