Journal of Gastroenterology

, Volume 54, Issue 5, pp 407–418 | Cite as

Deletion of IP3R1 by Pdgfrb-Cre in mice results in intestinal pseudo-obstruction and lethality

  • Hong Wang
  • Ran Jing
  • Christa Trexler
  • Yali Li
  • Huayuan Tang
  • Zhixiang Pan
  • Siting Zhu
  • Beili Zhao
  • Xi Fang
  • Jie Liu
  • Ju ChenEmail author
  • Kunfu OuyangEmail author
Original Article—Alimentary Tract



Inositol 1,4,5-trisphosphate receptors (IP3Rs) are a family of intracellular Ca2+ release channels located on the membrane of endoplasmic reticulum, which have been shown to play critical roles in various cellular and physiological functions. However, their function in regulating gastrointestinal (GI) tract motility in vivo remains unknown. Here, we investigated the physiological function of IP3R1 in the GI tract using genetically engineered mouse models.


Pdgfrb-Cre mice were bred with homozygous Itpr1 floxed (Itpr1f/f) mice to generate conditional IP3R1 knockout (pcR1KO) mice. Cell lineage tracing was used to determine where Pdgfrb-Cre-mediated gene deletion occurred in the GI tract. Isometric tension recording was used to measure the effects of IP3R1 deletion on muscle contraction.


In the mouse GI tract, Itpr1 gene deletion by Pdgfrb-Cre occurred in smooth muscle cells, enteric neurons, and interstitial cells of Cajal. pcR1KO mice developed impaired GI motility, with prolonged whole-gut transit time and abdominal distention. pcR1KO mice also exhibited lethality as early as 8 weeks of age and 50% of pcR1KO mice were dead by 40 weeks after birth. The frequency of spontaneous contractions in colonic circular muscles was dramatically decreased and the amplitude of spontaneous contractions was increased in pcR1KO mice. Deletion of IP3R1 in the GI tract also reduced the contractile response to the muscarinic agonist, carbachol, as well as to electrical field stimulation. However, KCl-induced contraction and expression of smooth muscle-specific contractile genes were not significantly altered in pcR1KO mice.


Here, we provided a novel mouse model for impaired GI motility and demonstrated that IP3R1 plays a critical role in regulating physiological function of GI tract in vivo.


IP3 receptor Ca2+ release channel Gut motility Intestinal pseudo-obstruction 



The work was supported by the National Science Foundation of China (31370823, 81700289, 31800767), the Guangdong Province Basic Research Foundation (2018A030310012), the Shenzhen Basic Research Foundation (KCYJ20160428154108239, KQJSCX20170330155020267, JCYJ20170818090044949, KQTD2015032709315529), and the National Institutes of Health (J.C. and F.X.). J.C. is the American Heart Association (AHA) Endowed Chair in Cardiovascular Research.

Author contribution

HW, RJ, YL, HT, ZP, SZ, and BZ performed research; KO, XF, and JC designed the research; KO, CT, XF, JL, and JC wrote the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Sanders KM, Koh SD, Ward SM. Interstitial cells of cajal as pacemakers in the gastrointestinal tract. Annu Rev Physiol. 2006;68:307–43.CrossRefGoogle Scholar
  2. 2.
    Knowles CH, Lindberg G, Panza E, et al. New perspectives in the diagnosis and management of enteric neuropathies. Nat Rev Gastroenterol Hepatol. 2013;10:206–18.CrossRefGoogle Scholar
  3. 3.
    Thompson WG, Longstreth GF, Drossman DA, et al. Functional bowel disorders and functional abdominal pain. Gut. 1999;45(Suppl 2):II43–7.Google Scholar
  4. 4.
    Keller J, Bassotti G, Clarke J, et al. Expert consensus document: advances in the diagnosis and classification of gastric and intestinal motility disorders. Nat Rev Gastroenterol Hepatol. 2018;15:291–308.CrossRefGoogle Scholar
  5. 5.
    Foskett JK, White C, Cheung KH, et al. Inositol trisphosphate receptor Ca2 + release channels. Physiol Rev. 2007;87:593–658.CrossRefGoogle Scholar
  6. 6.
    Nakazawa M, Uchida K, Aramaki M, et al. Inositol 1,4,5-trisphosphate receptors are essential for the development of the second heart field. J Mol Cell Cardiol. 2011;51:58–66.CrossRefGoogle Scholar
  7. 7.
    Uchida K, Aramaki M, Nakazawa M, et al. Gene knock-outs of inositol 1,4,5-trisphosphate receptors types 1 and 2 result in perturbation of cardiogenesis. PLoS ONE. 2010;5:e12500. CrossRefGoogle Scholar
  8. 8.
    Matsumoto M, Nakagawa T, Inoue T, et al. Ataxia and epileptic seizures in mice lacking type 1 inositol 1,4,5-trisphosphate receptor. Nature. 1996;379:168–71.CrossRefGoogle Scholar
  9. 9.
    Futatsugi A, Nakamura T, Yamada MK, et al. IP3 receptor types 2 and 3 mediate exocrine secretion underlying energy metabolism. Science. 2005;309:2232–4.CrossRefGoogle Scholar
  10. 10.
    Lin Q, Zhao G, Fang X, et al. IP3 receptors regulate vascular smooth muscle contractility and hypertension. JCI Insight. 2016. Scholar
  11. 11.
    Ouyang K, Leandro Gomez-Amaro R, Stachura DL, et al. Loss of IP3R-dependent Ca2 + signalling in thymocytes leads to aberrant development and acute lymphoblastic leukemia. Nat Commun. 2014. Scholar
  12. 12.
    Tang H, Wang H, Lin Q, et al. Loss of IP3 receptor-mediated Ca(2 +) release in mouse B cells results in abnormal B cell development and function. J Immunol. 2017;199:570–80.CrossRefGoogle Scholar
  13. 13.
    Iino S, Horiguchi K. Interstitial cells of cajal are involved in neurotransmission in the gastrointestinal tract. Acta Histochem Cytochem. 2006;39:145–53.CrossRefGoogle Scholar
  14. 14.
    Makhlouf GM, Murthy KS. Signal transduction in gastrointestinal smooth muscle. Cell Signal. 1997;9:269–76.CrossRefGoogle Scholar
  15. 15.
    Kuemmerle JF, Murthy KS, Makhlouf GM. Longitudinal smooth muscle of the mammalian intestine. A model for Ca2 + signaling by cADPR. Cell Biochem Biophys. 1998;28:31–44.CrossRefGoogle Scholar
  16. 16.
    Ward SM, Baker SA, de Faoite A, et al. Propagation of slow waves requires IP3 receptors and mitochondrial Ca2 + uptake in canine colonic muscles. J Physiol. 2003;549:207–18.CrossRefGoogle Scholar
  17. 17.
    Zhu MH, Sung TS, O’Driscoll K, et al. Intracellular Ca(2 +) release from endoplasmic reticulum regulates slow wave currents and pacemaker activity of interstitial cells of Cajal. Am J Physiol Cell Physiol. 2015;308:C608–20.CrossRefGoogle Scholar
  18. 18.
    Rehn M, Bader S, Bell A, et al. Distribution of voltage-dependent and intracellular Ca2 + channels in submucosal neurons from rat distal colon. Cell Tissue Res. 2013;353:355–66.CrossRefGoogle Scholar
  19. 19.
    Li X, Zima AV, Sheikh F, et al. Endothelin-1-induced arrhythmogenic Ca2 + signaling is abolished in atrial myocytes of inositol-1,4,5-trisphosphate(IP3)-receptor type 2-deficient mice. Circ Res. 2005;96:1274–81.CrossRefGoogle Scholar
  20. 20.
    Cooley N, Ouyang K, McMullen JR, et al. No contribution of IP3-R(2) to disease phenotype in models of dilated cardiomyopathy or pressure overload hypertrophy. Circ Heart Fail. 2013;6:318–25.CrossRefGoogle Scholar
  21. 21.
    Sato-Miyaoka M, Hisatsune C, Ebisui E, et al. Regulation of hair shedding by the type 3 IP3 receptor. J Invest Dermatol. 2012;132:2137–47.CrossRefGoogle Scholar
  22. 22.
    Suzuki H, Takano H, Yamamoto Y, et al. Properties of gastric smooth muscles obtained from mice which lack inositol trisphosphate receptor. J Physiol. 2000;525(Pt 1):105–11.CrossRefGoogle Scholar
  23. 23.
    Takano H, Imaeda K, Yamamoto Y, et al. Mechanical responses evoked by nerve stimulation in gastric muscles of mouse lacking inositol trisphosphate receptor. Auton Neurosci. 2001;87:249–57.CrossRefGoogle Scholar
  24. 24.
    Foo SS, Turner CJ, Adams S, et al. Ephrin-B2 controls cell motility and adhesion during blood-vessel-wall assembly. Cell. 2006;124:161–73.CrossRefGoogle Scholar
  25. 25.
    Fang X, Stroud MJ, Ouyang K, et al. Adipocyte-specific loss of PPARgamma attenuates cardiac hypertrophy. JCI Insight. 2016. Scholar
  26. 26.
    Lange S, Ouyang K, Meyer G, et al. Obscurin determines the architecture of the longitudinal sarcoplasmic reticulum. J Cell Sci. 2009;122:2640–50.CrossRefGoogle Scholar
  27. 27.
    Tamada H, Kiyama H. Suppression of c-Kit signaling induces adult neurogenesis in the mouse intestine after myenteric plexus ablation with benzalkonium chloride. Sci Rep. 2016. Scholar
  28. 28.
    Friebe A, Mergia E, Dangel O, et al. Fatal gastrointestinal obstruction and hypertension in mice lacking nitric oxide-sensitive guanylyl cyclase. Proc Natl Acad Sci U S A. 2007;104:7699–704.CrossRefGoogle Scholar
  29. 29.
    Srinivas S, Watanabe T, Lin CS, et al. Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus. BMC Dev Biol. 2001. Scholar
  30. 30.
    Mericskay M, Blanc J, Tritsch E, et al. Inducible mouse model of chronic intestinal pseudo-obstruction by smooth muscle-specific inactivation of the SRF gene. Gastroenterology. 2007;133:1960–70.CrossRefGoogle Scholar
  31. 31.
    Angstenberger M, Wegener JW, Pichler BJ, et al. Severe intestinal obstruction on induced smooth muscle-specific ablation of the transcription factor SRF in adult mice. Gastroenterology. 2007;133:1948–59.CrossRefGoogle Scholar
  32. 32.
    Parish IA, Stamp LA, Lorenzo AM, et al. A novel mutation in nucleoporin 35 causes murine degenerative colonic smooth muscle myopathy. Am J Pathol. 2016;186:2254–61.CrossRefGoogle Scholar
  33. 33.
    Niessen P, Rensen S, van Deursen J, et al. Smoothelin-a is essential for functional intestinal smooth muscle contractility in mice. Gastroenterology. 2005;129:1592–601.CrossRefGoogle Scholar
  34. 34.
    Qin X, Liu S, Lu Q, et al. Heterotrimeric G stimulatory protein alpha subunit is required for intestinal smooth muscle contraction in mice. Gastroenterology. 2017;152(1114–25):e5.Google Scholar
  35. 35.
    Isozaki K, Hirota S, Miyagawa J, et al. Deficiency of c-kit + cells in patients with a myopathic form of chronic idiopathic intestinal pseudo-obstruction. Am J Gastroenterol. 1997;92:332–4.Google Scholar
  36. 36.
    Boeckxstaens GE, Rumessen JJ, de Wit L, et al. Abnormal distribution of the interstitial cells of cajal in an adult patient with pseudo-obstruction and megaduodenum. Am J Gastroenterol. 2002;97:2120–6.CrossRefGoogle Scholar
  37. 37.
    Feldstein AE, Miller SM, El-Youssef M, et al. Chronic intestinal pseudoobstruction associated with altered interstitial cells of cajal networks. J Pediatr Gastroenterol Nutr. 2003;36:492–7.CrossRefGoogle Scholar
  38. 38.
    Santer RM. Survival of the population of NADPH-diaphorase stained myenteric neurons in the small intestine of aged rats. J Auton Nerv Syst. 1994;49:115–21.CrossRefGoogle Scholar
  39. 39.
    Fu M, Landreville S, Agapova OA, et al. Retinoblastoma protein prevents enteric nervous system defects and intestinal pseudo-obstruction. J Clin Invest. 2013;123:5152–64.CrossRefGoogle Scholar
  40. 40.
    Stenzel D, Nye E, Nisancioglu M, et al. Peripheral mural cell recruitment requires cell-autonomous heparan sulfate. Blood. 2009;114:915–24.CrossRefGoogle Scholar
  41. 41.
    Henderson NC, Arnold TD, Katamura Y, et al. Targeting of alphav integrin identifies a core molecular pathway that regulates fibrosis in several organs. Nat Med. 2013;19:1617–24.CrossRefGoogle Scholar
  42. 42.
    Klotz L, Norman S, Vieira JM, et al. Cardiac lymphatics are heterogeneous in origin and respond to injury. Nature. 2015;522:62–7.CrossRefGoogle Scholar
  43. 43.
    Stanczuk L, Martinez-Corral I, Ulvmar MH, et al. cKit lineage hemogenic endothelium-derived cells contribute to mesenteric lymphatic vessels. Cell Rep. 2015. Scholar
  44. 44.
    Owens GK, Kumar MS, Wamhoff BR. Molecular regulation of vascular smooth muscle cell differentiation in development and disease. Physiol Rev. 2004;84:767–801.CrossRefGoogle Scholar
  45. 45.
    Murthy KS, Grider JR, Makhlouf GM. InsP3-dependent Ca2 + mobilization in circular but not longitudinal muscle cells of intestine. Am J Physiol. 1991;261:G937–44.Google Scholar
  46. 46.
    Grider JR, Makhlouf GM. Suppression of inhibitory neural input to colonic circular muscle by opioid peptides. J Pharmacol Exp Ther. 1987;243:205–10.Google Scholar
  47. 47.
    Siefjediers A, Hardt M, Prinz G, et al. Characterization of inositol 1,4,5-trisphosphate (IP3) receptor subtypes at rat colonic epithelium. Cell Calcium. 2007;41:303–15.CrossRefGoogle Scholar
  48. 48.
    Malysz J, Donnelly G, Huizinga JD. Regulation of slow wave frequency by IP(3)-sensitive calcium release in the murine small intestine. Am J Physiol Gastrointest Liver Physiol. 2001;280:G439–48.CrossRefGoogle Scholar
  49. 49.
    Ward SM, Ordog T, Koh SD, et al. Pacemaking in interstitial cells of Cajal depends upon calcium handling by endoplasmic reticulum and mitochondria. J Physiol. 2000;525(Pt 2):355–61.CrossRefGoogle Scholar
  50. 50.
    Dickens EJ, Edwards FR, Hirst GD. Vagal inhibition in the antral region of guinea pig stomach. Am J Physiol Gastrointest Liver Physiol. 2000;279:G388–99.CrossRefGoogle Scholar

Copyright information

© Japanese Society of Gastroenterology 2018

Authors and Affiliations

  1. 1.Drug Discovery Center, State Key Laboratory of Chemical Oncogenomics, School of Chemical Biology and BiotechnologyPeking University Shenzhen Graduate SchoolShenzhenChina
  2. 2.Xiangya HospitalCentral South UniversityChangshaChina
  3. 3.Department of MedicineUniversity of California-San DiegoLa JollaUSA
  4. 4.Department of Pathophysiology, School of MedicineShenzhen UniversityShenzhenChina

Personalised recommendations