Functional properties of submucosal venules in the rat stomach

  • Retsu MitsuiEmail author
  • Hikaru Hashitani
Muscle physiology


Venules in the stomach may have intrinsic properties for maintaining active microcirculation drainage even during gastric filling. Properties of spontaneous and nerve-mediated activity of submucosal venules in the rat stomach were investigated. Changes in vasodiameter and intracellular Ca2+ in venular smooth muscle cells (SMCs) were monitored by video tracking and Fluo-8 Ca2+ imaging, respectively. Venular SMCs developed synchronous spontaneous Ca2+ transients and corresponding rhythmic constrictions of the venules. Nominally Ca2+-free solution or an L-type Ca2+ channel blocker (1 μM nifedipine) disrupted the Ca2+ transient synchrony and abolished spontaneous constrictions. Spontaneous constrictions were also prevented by inhibitors of sarcoplasmic reticulum Ca2+-ATPase (10 μM cyclopiazonic acid (CPA)), IP3 receptors (100 μM 2-APB) or Ca2+-activated Cl channels (100 μM niflumic acid). Transmural nerve stimulation (TNS) induced a long-lasting venular constriction that was abolished by α-adrenoceptor antagonist (1 μM phentolamine), while TNS evoked a sympathetic transient constriction of arterioles that was abolished by a combination of phentolamine and a P2 purinoceptor antagonist (10 μM pyridoxal-phosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS)). Consistently, P2X1 purinoceptor immunoreactivity was detected in arteriolar but not venular SMCs. Primary afferent nerve stimulation (300 nM capsaicin) caused a venular dilatation by releasing calcitonin gene-related peptide. Thus, Ca2+ release from the sarcoplasmic reticulum may play a fundamental role in the generation of spontaneous Ca2+ transients, while electrical coupling amongst venular SMCs via L-type Ca2+ channel activation appears to be critical for Ca2+ transient synchrony as well as spontaneous contractions. Sympathetic venular constrictions appear to be exclusively mediated by noradrenaline due to the lack of P2X1 receptor in venular SMCs.


Microvasculature Smooth muscle Vasomotion Autonomic nerves Digestive tract 



The authors wish to thank Dr. Richard Lang (Monash University) for critical reading of the manuscript. This study was supported by Grant-in-Aid for Research in Nagoya City University to R.M., a research grant from Ichihara International Scholarship Foundation to R.M., Grant-in-Aid for Scientific Research (B) (No. 22390304) from Japan Society for Promotion of the Science (JSPS) to H.H., Grant-in-Aid for Challenging Exploratory Research (No. 21659377) from JSPS to H.H. and Grant-in-Aid for Young Scientists (B) (No. 26860521) from JSPS to R.M..


The experimental protocols in the present study were approved by the Nagoya City University Medical School experimental animal committee.

Conflict of interest

The authors have no conflict of interest.


  1. 1.
    Andersson PO (1984) Vascular control in the colon and rectum. Scand J Gastroenterol Suppl 93:65–78PubMedGoogle Scholar
  2. 2.
    Bultynck G, Sienaert I, Parys JB, Callewaert G, De Smedt H, Boens N, Dehaen W, Missiaen L (2003) Pharmacology of inositol trisphosphate receptors. Pflugers Arch 445:629–642PubMedGoogle Scholar
  3. 3.
    Chen RY, Li DS, Guth PH (1992) Role of calcitonin gene-related peptide in capsaicin-induced gastric submucosal arteriolar dilation. Am J Physiol 262:H1350–H1355PubMedGoogle Scholar
  4. 4.
    De Fontgalland D, Wattchow DA, Costa M, Brookes SJ (2008) Immunohistochemical characterization of the innervation of human colonic mesenteric and submucosal blood vessels. Neurogastroenterol Motil 20:1212–1226CrossRefPubMedGoogle Scholar
  5. 5.
    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 106:242–249CrossRefPubMedCentralPubMedGoogle Scholar
  6. 6.
    Figini M, Emanueli C, Grady EF, Kirkwood K, Payan DG, Ansel J, Gerard C, Geppetti P, Bunnett N (1997) Substance P and bradykinin stimulate plasma extravasation in the mouse gastrointestinal tract and pancreas. Am J Physiol 272:G785–G793PubMedGoogle Scholar
  7. 7.
    Folkow B, Lewis DH, Lundgren O, Mellander S, Wallentin I (1964) The effect of graded vasoconstrictor fibre stimulation on the intestinal resistance and capacitance vessels. Acta Physiol Scand 61:445–457PubMedGoogle Scholar
  8. 8.
    Furness JB, Marshall JM (1974) Correlation of the directly observed responses of mesenteric vessels of the rat to nerve stimulation and noradrenaline with the distribution of adrenergic nerves. J Physiol 239:75–88CrossRefPubMedCentralPubMedGoogle Scholar
  9. 9.
    Galligan JJ, Herring A, Harpstead T (1995) Pharmacological characterization of purinoceptor-mediated constriction of submucosal arterioles in guinea pig ileum. J Pharmacol Exp Ther 274:1425–1430PubMedGoogle Scholar
  10. 10.
    Gomez-Pinilla PJ, Gibbons SJ, Bardsley MR, Lorincz A, Pozo MJ, Pasricha PJ, Van de Rijn M, West RB, Sarr MG, Kendrick ML, Cima RR, Dozois EJ, Larson DW, Ordog T, Farrugia G (2009) Ano1 is a selective marker of interstitial cells of Cajal in the human and mouse gastrointestinal tract. Am J Physiol Gastrointest Liver Physiol 296:G1370–G1381CrossRefPubMedCentralPubMedGoogle Scholar
  11. 11.
    Guth PH, Smith E (1975) Neural control of gastric mucosal blood flow in the rat. Gastroenterology 69:935–940PubMedGoogle Scholar
  12. 12.
    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 545:615–627CrossRefPubMedCentralPubMedGoogle Scholar
  13. 13.
    Hashitani H, Edwards FR (1999) Spontaneous and neurally activated depolarizations in smooth muscle cells of the guinea-pig urethra. J Physiol 514:459–470CrossRefPubMedCentralPubMedGoogle Scholar
  14. 14.
    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 167:1723–1736CrossRefPubMedCentralPubMedGoogle 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.
    Hirose K, Iino M, Endo M (1993) Caffeine inhibits Ca2+-mediated potentiation of inositol 1,4,5-trisphosphate-induced Ca2+ release in permeabilized vascular smooth muscle cells. Biochem Biophys Res Commun 194:726–732CrossRefPubMedGoogle Scholar
  17. 17.
    Hirst GD (1977) Neuromuscular transmission in arterioles of guinea-pig submucosa. J Physiol 273:263–275CrossRefPubMedCentralPubMedGoogle Scholar
  18. 18.
    Hirst GD, Jobling P (1989) The distribution of gamma-adrenoceptors and P2 purinoceptors in mesenteric arteries and veins of the guinea-pig. Br J Pharmacol 96:993–999CrossRefPubMedCentralPubMedGoogle Scholar
  19. 19.
    Holzer P, Livingston EH, Guth PH (1991) Sensory neurons signal for an increase in rat gastric mucosal blood flow in the face of pending acid injury. Gastroenterology 101:416–423PubMedGoogle Scholar
  20. 20.
    Iino M (1989) Calcium-induced calcium release mechanism in guinea pig taenia caeci. J Gen Physiol 94:363–383CrossRefPubMedGoogle Scholar
  21. 21.
    Johnstone S, Isakson B, Locke D (2009) Biological and biophysical properties of vascular connexin channels. Int Rev Cell Mol Biol 278:69–118CrossRefPubMedCentralPubMedGoogle Scholar
  22. 22.
    Kotecha N, Neild TO (1995) Actions of vasodilator nerves on arteriolar smooth muscle and neurotransmitter release from sympathetic nerves in the guinea-pig small intestine. J Physiol 489:849–855CrossRefPubMedCentralPubMedGoogle Scholar
  23. 23.
    Kunisawa Y, Komuro T (2008) Interstitial cells of Cajal associated with the submucosal plexus of the guinea-pig stomach. Neurosci Lett 434:273–276CrossRefPubMedGoogle Scholar
  24. 24.
    Lamont C, Vial C, Evans RJ, Wier WG (2006) P2X1 receptors mediate sympathetic postjunctional Ca2+ transients in mesenteric small arteries. Am J Physiol Heart Circ Physiol 291:H3106–H3113CrossRefPubMedGoogle Scholar
  25. 25.
    Mazzone A, Eisenman ST, Strege PR, Yao Z, Ordog T, Gibbons SJ, Farrugia G (2012) Inhibition of cell proliferation by a selective inhibitor of the Ca2+-activated Cl channel, Ano1. Biochem Biophys Res Commun 427:248–253CrossRefPubMedCentralPubMedGoogle 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, Miyamoto S, Takano H, Hashitani H (2013) Properties of submucosal venules in the rat distal colon. Br J Pharmacol 170:968–977CrossRefPubMedCentralPubMedGoogle Scholar
  28. 28.
    Morgan KG (1983) Comparison of membrane electrical activity of cat gastric submucosal arterioles and venules. J Physiol 345:135–147CrossRefPubMedCentralPubMedGoogle Scholar
  29. 29.
    Neild TO (1989) Measurement of arteriole diameter changes by analysis of television images. Blood Vessels 26:48–52PubMedGoogle Scholar
  30. 30.
    Neild TO, Shen KZ, Surprenant A (1990) Vasodilatation of arterioles by acetylcholine released from single neurones in the guinea-pig submucosal plexus. J Physiol 420:247–265CrossRefPubMedCentralPubMedGoogle Scholar
  31. 31.
    Nilsson H, Aalkjær C (2003) Vasomotion: mechanisms and physiological importance. Mol Interv 3:79–89CrossRefPubMedGoogle Scholar
  32. 32.
    Parker I, Ivorra I (1991) Caffeine inhibits inositol trisphosphate-mediated liberation of intracellular calcium in Xenopus oocytes. J Physiol 433:229–240CrossRefPubMedCentralPubMedGoogle Scholar
  33. 33.
    Peng H, Matchkov V, Ivarsen A, Aalkjær C, Nilsson H (2001) Hypothesis for the initiation of vasomotion. Circ Res 88:810–815CrossRefPubMedGoogle Scholar
  34. 34.
    Ralevic V (2009) Purines as neurotransmitters and neuromodulators in blood vessels. Curr Vasc Pharmacol 7:3–14CrossRefPubMedGoogle Scholar
  35. 35.
    Reed DE, Vanner SJ (2003) Long vasodilator reflexes projecting through the myenteric plexus in guinea-pig ileum. J Physiol 553:911–924CrossRefPubMedCentralPubMedGoogle Scholar
  36. 36.
    Sergeant GP, Hollywood MA, McCloskey KD, McHale NG, Thornbury KD (2001) Role of IP3 in modulation of spontaneous activity in pacemaker cells of rabbit urethra. Am J Physiol Cell Physiol 280:C1349–C1356PubMedGoogle Scholar
  37. 37.
    Shimizu Y, Mochizuki S, Mitsui R, Hashitani H (2014) Neurohumoral regulation of spontaneous constrictions in suburothelial venules of the rat urinary bladder. Vascul Pharmacol 60:84–94CrossRefPubMedGoogle Scholar
  38. 38.
    Sternini C, Reeve JR Jr, Brecha N (1987) Distribution and characterization of calcitonin gene-related peptide immunoreactivity in the digestive system of normal and capsaicin-treated rats. Gastroenterology 93:852–862PubMedGoogle Scholar
  39. 39.
    Su HC, Bishop AE, Power RF, Hamada Y, Polak JM (1987) Dual intrinsic and extrinsic origins of CGRP- and NPY-immunoreactive nerves of rat gut and pancreas. J Neurosci 7:2674–2687PubMedGoogle Scholar
  40. 40.
    Suzuki H (1981) Effects of endogenous and exogenous noradrenaline on the smooth muscle of guinea-pig mesenteric vein. J Physiol 321:495–512CrossRefPubMedCentralPubMedGoogle Scholar
  41. 41.
    Tamada H, Komuro T (2012) Ultrastructural characterization of interstitial cells of Cajal associated with the submucosal plexus in the proximal colon of the guinea pig. Cell Tissue Res 347:319–326CrossRefPubMedGoogle Scholar
  42. 42.
    Vanner S (1994) Corelease of neuropeptides from capsaicin-sensitive afferents dilates submucosal arterioles in guinea pig ileum. Am J Physiol 267:G650–G655PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

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

Personalised recommendations