title: Type 3 muscarinic acetylcholine receptor stimulation is a determinant of endothelial barrier function and adherens junctions integrity: role of protein-tyrosine phosphatase 1B.
abstract: The main purpose of this study was to investigate whether type 3 muscarinic acetylcholine receptor (M3R) dysfunction induced vascular hyperpermeability. Transwell system analysis showed that M3R inhibition by selective antagonist 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP) and small interfering RNA both increased endothelial permeability. Using coimmunoprecipitation and Western blot assay, we found that M3R inhibition increased VE-cadherin and β-catenin tyrosine phosphorylation without affecting their expression. Using PTP1B siRNA, we found that PTP1B was required for maintaining VE-cadherin and β-catenin protein dephosphorylation. In addition, 4-DAMP suppressed PTP1B activity by reducing cyclic adenosine monophosphate (cAMP), but not protein kinase Cα (PKCα). These data indicate that M3R preserves the endothelial barrier function through a mechanism potentially maintaining PTP1B activity, keeping the adherens junction proteins (AJPs) dephosphorylation.
authors.affiliation: Department of Cardiovascular Surgery, Xiehe Hospital, Huazhong University of Science and Technology, Wuhan 430022; Department of Cardiovascular Surgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
authors.forename: Zhou-Yang
authors.initials: ZY
authors.lastname: Jiao
authors.affiliation: Department of Pediatrics, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Wu
authors.affiliation: Department of Cardiovascular Surgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Liu
authors.affiliation: Department of Cardiovascular Surgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
authors.forename: Bing
authors.initials: B
authors.lastname: Wen
authors.affiliation: Department of Cardiovascular Surgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
authors.forename: Wen-Zeng
authors.initials: WZ
authors.lastname: Zhao
authors.affiliation: Department of Cardiovascular Surgery, Xiehe Hospital, Huazhong University of Science and Technology, Wuhan 430022, China.
authors.forename: Xin-Ling
authors.initials: XL
authors.lastname: Du
mesh_terms: D022005:Adherens Junctions / Q000378:metabolism*; D015703:Antigens, CD / Q000378:metabolism; D015820:Cadherins / Q000378:metabolism; D002463:Cell Membrane Permeability; D000242:Cyclic AMP / Q000378:metabolism; D004730:Endothelium, Vascular / Q000378:metabolism*; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism; D006801:Humans; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D051571:Protein Kinase C-alpha / Q000378:metabolism; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1 / Q000378:metabolism*; D043587:Receptor, Muscarinic M3; D011976:Receptors, Muscarinic / Q000378:metabolism*; D051176:beta Catenin / Q000378:metabolism; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015820:Cadherins; D000242:Cyclic AMP; D019000:Phosphotyrosine; D051571:Protein Kinase C-alpha; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; D043587:Receptor, Muscarinic M3; D011976:Receptors, Muscarinic; D051176:beta Catenin; D000099152:Cadherin 5; C000595896:CHRM3 protein, human; C516607:PTPN1 protein, human