Abstract
Submandibular gland (SMG) autotransplantation is an effective therapy for treating severe dry eye syndrome. However, epiphora occurs in more than 40% of patients 6 months after operation. We previously found that muscarinic acetylcholine receptor (mAChR) plays a crucial role in regulating SMG secretion partially through the modulation on tight junction (TJ)-based paracellular pathway. Therefore, the present study aimed to investigate the possible involvement of mAChR and TJ in a rabbit long-term model of SMG transplantation. We found that SMG secretion was significantly increased on postoperative days 90 and 180, which imitated epiphora observed in the patients with SMG transplantation. Although the mRNA expression and fluorescence intensity of M1 and M3 mAChR subtypes were reversed to control levels on postoperative days 30, 90, and 180, the content of β-arrestin2, but not β-arrestin1, was gradually decreased after transplantation, which suggests that mAChR may be hypersensitive in late phase of SMG transplantation. The width of acinar TJs was enlarged and fluorescence intensity of F-actin in peri-apicolateral membranes were remarkably increased on postoperative days 90 and 180. Topical treatment with atropine gel significantly reduced SMG secretion, TJ width, as well as F-actin intensity in peri-apicolateral membranes on postoperative days 180. Moreover, in a perfused rabbit SMG, carbachol increased salivary secretion, enlarged TJ width, and induced F-actin rearrangement, whereas these responses were inhibited by atropine pretreatment. Taken together, our findings suggest that the hypersensitive mAChR may contribute to epiphora in late phase of SMG transplantation through modulating TJ-based paracellular permeability.
Similar content being viewed by others
References
Abe A, Takano K, Kojima T, Nomura K, Kakuki T, Kaneko Y, Yamamoto M, Takahashi H, Himi T (2016) Interferon-gamma increased epithelial barrier function via upregulating claudin-7 expression in human submandibular gland duct epithelium. J Mol Histol 47:353–363. doi:10.1007/s10735-016-9667-2
Baum BJ (1993) Principles of saliva secretion. Ann N. Y. Acad Sci 694:17–23
Bruewer M, Hopkins AM, Hobert ME, Nusrat A, Madara JL (2004) RhoA, Rac1, and Cdc42 exert distinct effects on epithelial barrier via selective structural and biochemical modulation of junctional proteins and F-actin. Am J Physiol Cell Physiol 287:C327–C335. doi:10.1152/ajpcell.00087.2004
Chung NP, Mruk D, Mo MY, Lee WM, Cheng CY (2001) A 22-amino acid synthetic peptide corresponding to the second extracellular loop of rat occludin perturbs the blood-testis barrier and disrupts spermatogenesis reversibly in vivo. Biol Reprod 65:1340–1351
Cong X, Zhang Y, Shi L et al (2012) Activation of transient receptor potential vanilloid subtype 1 increases expression and permeability of tight junction in normal and hyposecretory submandibular gland. Lab Invest 92:753–768. doi:10.1038/labinvest.2012.12
Cong X, Zhang Y, Yang NY et al (2013) Occludin is required for TRPV1-modulated paracellular permeability in the submandibular gland. J Cell Sci 126:1109–1121. doi:10.1242/jcs.111781
Cong X, Zhang Y, Li J et al (2015) Claudin-4 is required for modulation of paracellular permeability by muscarinic acetylcholine receptor in epithelial cells. J Cell Sci 128:2271–2286. doi:10.1242/jcs.165878
Ding C, Cong X, Zhang Y, Yang NY, Li SL, Wu LL, Yu GY (2014) Hypersensitive mAChRs are involved in the epiphora of transplanted glands. J Dent Res 93:306–312. doi:10.1177/0022034513519107
Ewert P, Aguilera S, Alliende C et al (2010) Disruption of tight junction structure in salivary glands from Sjogren’s syndrome patients is linked to proinflammatory cytokine exposure. Arthritis Rheum 62:1280–1289. doi:10.1002/art.27362
Fleegal MA, Hom S, Borg LK, Davis TP (2005) Activation of PKC modulates blood-brain barrier endothelial cell permeability changes induced by hypoxia and posthypoxic reoxygenation. Am J Physiol Heart Circ Physiol 289:H2012–H2019. doi:10.1152/ajpheart.00495.2005
Furuse M, Hata M, Furuse K et al (2002) Claudin-based tight junctions are crucial for the mammalian epidermal barrier: a lesson from claudin-1-deficient mice. J Cell Biol 156:1099–1111. doi:10.1083/jcb.200110122
Gautam D, Heard TS, Cui Y, Miller G, Bloodworth L, Wess J (2004) Cholinergic stimulation of salivary secretion studied with M1 and M3 muscarinic receptor single- and double-knockout mice. Mol Pharmacol 66:260–267. doi:10.1124/mol.66.2.260
Geerling G, Sieg P, Bastian GO, Laqua H (1998) Transplantation of the autologous submandibular gland for most severe cases of keratoconjunctivitis sicca. Ophthalmology 105:327–335
Geerling G, Garrett JR, Paterson KL et al (2008) Innervation and secretory function of transplanted human submandibular salivary glands. Transplantation 85:135–140. doi:10.1097/01.tp.0000296060.39823.a4
Gonzalez-Mariscal L, Betanzos A, Nava P, Jaramillo BE (2003) Tight junction proteins. Prog Biophys Mol Biol 81:1–44
Iwamoto N, Higashi T, Furuse M (2014) Localization of angulin-1/LSR and tricellulin at tricellular contacts of brain and retinal endothelial cells in vivo. Cell Struct Funct 39:1–8
Kilkenny C, Browne WJ, Cuthill IC, Emerson M, Altman DG (2010) Improving bioscience research reporting: The ARRIVE guidelines for reporting animal research. J Pharmacol Pharmacother 1:94–99. doi:10.4103/0976-500X.72351
Krupnick JG, Benovic JL (1998) The role of receptor kinases and arrestins in G protein-coupled receptor regulation. Annu Rev Pharmacol Toxicol 38:289–319. doi:10.1146/annurev.pharmtox.38.1.289
Kuo TM, Luo SY, Chiang SL et al (2014) Arecoline induces TNF-alpha production and Zonula Occludens-1 redistribution in mouse Sertoli TM4 cells. J Biomed Sci 21:93. doi:10.1186/s12929-014-0093-z
Lopez-Jornet P, Camacho-Alonso F, Bermejo-Fenoll A (2006) A simple test for salivary gland hypofunction using oral Schirmer’s test. J Oral Pathol Med 35:244–248. doi:10.1111/j.1600-0714.2006.00411.x
Luo J, Busillo JM, Benovic JL (2008) M3 muscarinic acetylcholine receptor-mediated signaling is regulated by distinct mechanisms. Mol Pharmacol 74:338–347. doi:10.1124/mol.107.044750
Melvin JE, Yule D, Shuttleworth T, Begenisich T (2005) Regulation of fluid and electrolyte secretion in salivary gland acinar cells. Annu Rev Physiol 67:445–469. doi:10.1146/annurev.physiol.67.041703.084745
Moore CA, Milano SK, Benovic JL (2007) Regulation of receptor trafficking by GRKs and arrestins. Annu Rev Physiol 69:451–482. doi:10.1146/annurev.physiol.69.022405.154712
Murakami M, Shachar-Hill B, Steward MC, Hill AE (2001) The paracellular component of water flow in the rat submandibular salivary gland. J Physiol 537:899–906
Murube-del-Castillo J (1986) Transplantation of salivary gland to the lacrimal basin. Scand J Rheumatol Suppl 61:264–267
Muto S, Hata M, Taniguchi J et al (2010) Claudin-2-deficient mice are defective in the leaky and cation-selective paracellular permeability properties of renal proximal tubules. Proc Natl Acad Sci USA 107:8011–8016. doi:10.1073/pnas.0912901107
Nighot PK, Blikslager AT (2010) ClC-2 regulates mucosal barrier function associated with structural changes to the villus and epithelial tight junction. Am J Physiol Gastrointest Liver Physiol 299:G449–G456. doi:10.1152/ajpgi.00520.2009
Scully C (2003) Drug effects on salivary glands: dry mouth. Oral Dis 9:165–176
Shi L, Cong X, Zhang Y et al (2010) Carbachol improves secretion in the early phase after rabbit submandibular gland transplantation. Oral Dis 16:351–359. doi:10.1111/j.1601-0825.2009.01633.x
Sieg P, Geerling G, Kosmehl H, Lauer I, Warnecke K, von Domarus H (2000) Microvascular submandibular gland transfer for severe cases of keratoconjunctivitis sicca. Plast Reconstr Surg 106:554–560. (discussion 561–552)
Steward MC, Seo Y, Rawlings JM, Case RM (1990) Water permeability of acinar cell membranes in the isolated perfused rabbit mandibular salivary gland. J Physiol 431:571–583
Su JZ, Liu XJ, Zhang L, Yu GY (2013) Schirmer test in transplanted submandibular gland: influencing factors and a modified measurement method. Cornea 32:419–422. doi:10.1097/ICO.0b013e31825aba2c
Su JZ, Cai ZG, Yu GY (2015) Microvascular autologous submandibular gland transplantation in severe cases of keratoconjunctivitis sicca. Maxillofac Plast Reconstr Surg 37:5. doi:10.1186/s40902-015-0006-4
Sumida T, Tsuboi H, Iizuka M, Asashima H, Matsumoto I (2013) Anti-M3 muscarinic acetylcholine receptor antibodies in patients with Sjogren’s syndrome. Mod Rheumatol 23:841–845. doi:10.1007/s10165-012-0788-5
Tamura A, Hayashi H, Imasato M et al (2011) Loss of claudin-15, but not claudin-2, causes Na+ deficiency and glucose malabsorption in mouse small intestine. Gastroenterology 140:913–923. doi:10.1053/j.gastro.2010.08.006
Tsukita S, Furuse M, Itoh M (2001) Multifunctional strands in tight junctions. Nat Rev Mol Cell Biol 2:285–293. doi:10.1038/35067088
Wiseman LR, Faulds D (1995) Oral pilocarpine: a review of its pharmacological properties and clinical potential in xerostomia. Drugs 49:143–155
Yang NY, Shi L, Zhang Y et al (2013) Ischemic preconditioning reduces transplanted submandibular gland injury. J Surg Res 179:e265–e273. doi:10.1016/j.jss.2012.02.066
Yu GY, Zhu ZH, Mao C et al (2004) Microvascular autologous submandibular gland transfer in severe cases of keratoconjunctivitis sicca. Int J Oral Maxillofac Surg 33:235–239. doi:10.1006/ijom.2002.0438
Zhang Y, Cong X, Shi L et al (2010) Activation of transient receptor potential vanilloid subtype 1 increases secretion of the hypofunctional, transplanted submandibular gland. Am J Physiol Gastrointest Liver Physiol 299:G54–G62. doi:10.1152/ajpgi.00528.2009
Zhang GH, Wu LL, Yu GY (2013) Tight junctions and paracellular fluid and ion transport in salivary glands. Chin J Dent Res 16:13–46
Zhang LW, Cong X, Zhang Y et al (2016) Interleukin-17 impairs salivary tight junction integrity in Sjogren’s syndrome. J Dent Res 95:784–792. doi:10.1177/0022034516634647
Acknowledgements
This study was supported by the National Natural Science Foundation of China 81470756 (NYY), 81300893 (XC), and 81400526 (GYY), and Beijing Natural Science Foundation 7154204 (NYY).
Author contributions
NYY performed the major experiments and wrote the manuscript. CD, JL, YZ and RLX were responsible for the isolation and perfusion of rabbit submandibular gland. LLW and GYY participated in data interpretation and manuscript improvement. XC designed the study, analyzed the data, and wrote the manuscript. All authors read and approved the final manuscript.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no competing interests.
Rights and permissions
About this article
Cite this article
Yang, NY., Ding, C., Li, J. et al. Muscarinic acetylcholine receptor-mediated tight junction opening is involved in epiphora in late phase of submandibular gland transplantation. J Mol Hist 48, 99–111 (2017). https://doi.org/10.1007/s10735-016-9709-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10735-016-9709-9