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Journal of Peking University(Health Sciences) ›› 2019, Vol. 51 ›› Issue (3): 390-396. doi: 10.19723/j.issn.1671-167X.2019.03.003

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Role and mechanism of muscarinic acetylcholine receptor in the regulation of submandibular gland secretion

Xin CONG1,2,Sai-nan MIN3,Li-ling WU1,2,Zhi-gang CAI3,Guang-yan YU3△()   

  1. 1. Center for Salivary Gland Diseases, Peking University School and Hospital of Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Laboratory for Digital and Material Technology of Stomatology & Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China
    2. Department of Physiology and Pathophysiology, Peking University School of Basic Medical Sciences, Beijing 100191, China
    3. Department of Oral and Maxillofacial Surgery, Peking University School and Hospital of Stomatology, Beijing 100081, China
  • Received:2019-03-31 Online:2019-06-18 Published:2019-06-26
  • Supported by:
    Supported by the National Natural Science Foundation of China(81671005、81771093)

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Abstract:

SUMMARY Muscarinic acetylcholine receptors (mAChRs), including M1-M5 subtypes, are classic receptors in regulating water, ion, and solute transport in salivary gland. Our work focuses on the studies on the expression pattern and function of mAChR in the submandibular gland (SMG), and the under-lying mechanism involved in the mAChR-regulated secretion, together with the effect of parasympathectomy on the salivary secretion. Microvascular autotransplantation of SMG into the temporal fossa provides a continuous and endogenous source of fluids, and is currently an effective method for treating severe keratoconjunctivitis sicca. By using RT-PCR, Western blotting, and immunofluorescence, our data demonstrated that the expression of M1 and M3 subtypes were decreased in latent period in rabbit SMG autotransplantation model, whereas carbachol stimulation promoted the salivary secretion, as well as M1 and M3 expressions. By contrast, mAChRs were hypersensitive in epiphora SMGs, whereas atropine gel and botulinum toxin A application significantly inhibited the hypersecretion in both animal models and patients. Furthermore, the possible intracellular signal molecules involved in the mAChR-modulated saliva-ry secretion were explored. Activation of mAChR upregulated the expression of aquaporin 5 (AQP5), the main transporter that mediated water secretion through transcellular pathway, and led to AQP5 trafficking from lipid rafts to non-lipid microdomain. Extracellular signal-regulated kinase 1/2 (ERK1/2) was involved in the mAChR-regulated AQP5 content. mAChR activation also modulated the expression, distribution, and function of tight junction proteins, and increased paracellular permeability. ERK1/2/β-arrestin2/clathrin/ubiquitin signaling pathway was responsible for the mAChR-regulated downregulation of tight junction molecule claudin-4. Cytoskeleton filamentous actin (F-actin) was also involved in the distribution and barrier function of epithelial tight junctions. Besides, endothelial tight junctions were opened by mAChR agonist-evoked salivation in the mice. Furthermore, parasympathetic denervation increased resting salivary secretion in the long terminrats and minipigs. Taken together, our work demonstrated that mAChR regulated saliva secretion via transcellular and paracellular pathways in SMG epithe-lium as well as tight junction opening in SMG endothelium. Modulation of mAChR might be a promising strategy to ameliorate SMG dysfunction.

Key words: Muscarinic acetylcholine receptor, Submandibular gland, Secretion, Aquaporin, Tight junction

CLC Number: 

  • R781.7

Figure 1

The effect of mAChR activation on material transport in submandibular gland epithelium and endothelium A, the effect of mAChR activation on transcellular and paracellular transport in acinar epithelial cells of submandibular gland; B, the effect of mAChR activation on the opening of endothelial tight junctions in submandibular gland. mAChR, muscarinic acetylcholine receptor; Pilo, pilocarpine; Cch, carbachol; Cevi, cevimeline; AQP5, aquaporin 5; ERK1/2, extracellular signal-regulated kinase 1/2; βARR2, β-arrestin 2; MLC2, myosin light chain 2; F-actin, filamentous actin."

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