Pro-Resolving Mediator Annexin A1 Regulates Intracellular Ca2+ and Mucin Secretion in Cultured Goblet Cells Suggesting a New Use in Inflammatory Conjunctival Diseases
The amount of mucin secreted by conjunctival goblet cells is regulated to ensure the optimal level for protection of the ocular surface. Under physiological conditions lipid specialized pro-resolving mediators (SPM) are essential for maintaining tissue homeostasis including the conjunctiva. The prot...
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doaj-b96e0ffaf4194d2ba21cc43ae435c0cd2021-04-22T06:48:40ZengFrontiers Media S.A.Frontiers in Immunology1664-32242021-04-011210.3389/fimmu.2021.618653618653Pro-Resolving Mediator Annexin A1 Regulates Intracellular Ca2+ and Mucin Secretion in Cultured Goblet Cells Suggesting a New Use in Inflammatory Conjunctival DiseasesAnne V. Lyngstadaas0Anne V. Lyngstadaas1Anne V. Lyngstadaas2Markus V. Olsen3Markus V. Olsen4Markus V. Olsen5Jeffrey A. Bair6Robin R. Hodges7Tor P. Utheim8Tor P. Utheim9Tor P. Utheim10Charles N. Serhan11Darlene A. Dartt12Schepens Eye Research institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA, United StatesInstitute of Clinical Medicine, Faculty of Medicine, University of Oslo, Oslo, NorwayDepartment of Medical Biochemistry, Oslo University Hospital, Oslo, NorwaySchepens Eye Research institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA, United StatesInstitute of Clinical Medicine, Faculty of Medicine, University of Oslo, Oslo, NorwayDepartment of Medical Biochemistry, Oslo University Hospital, Oslo, NorwaySchepens Eye Research institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA, United StatesSchepens Eye Research institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA, United StatesSchepens Eye Research institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA, United StatesDepartment of Medical Biochemistry, Oslo University Hospital, Oslo, NorwayDepartment of Plastic and Reconstructive Surgery, University of Oslo, Oslo, NorwayCenter for Experimental Therapeutics and Reperfusion Injury, Department of Anesthesia, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA, United StatesSchepens Eye Research institute/Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, MA, United StatesThe amount of mucin secreted by conjunctival goblet cells is regulated to ensure the optimal level for protection of the ocular surface. Under physiological conditions lipid specialized pro-resolving mediators (SPM) are essential for maintaining tissue homeostasis including the conjunctiva. The protein Annexin A1 (AnxA1) can act as an SPM. We used cultured rat conjunctival goblet cells to determine if AnxA1 stimulates an increase in intracellular [Ca2+] ([Ca2+]i) and mucin secretion and to identify the signaling pathways. The increase in [Ca2+]i was determined using fura2/AM and mucin secretion was measured using an enzyme-linked lectin assay. AnxA1 stimulated an increase in [Ca2+]i and mucin secretion that was blocked by the cell-permeant Ca2+ chelator BAPTA/AM and the ALX/FPR2 receptor inhibitor BOC2. AnxA1 increased [Ca2+]i to a similar extent as the SPMs lipoxin A4 and Resolvin (Rv) D1 and histamine. The AnxA1 increase in [Ca2+]i and mucin secretion were inhibited by blocking the phospholipase C (PLC) pathway including PLC, the IP3 receptor, the Ca2+/ATPase that causes the intracellular Ca2+ stores to empty, and blockade of Ca2+ influx. Inhibition of protein kinase C (PKC) and Ca2+/calmodulin-dependent protein kinase also decreased the AnxA1-stimulated increase in [Ca2+]i and mucin secretion. In contrast inhibitors of ERK 1/2, phospholipase A2 (PLA2), and phospholipase D (PLD) did not alter AnxA1-stimulated increase in [Ca2+]i, but did inhibit mucin secretion. Activation of protein kinase A did not decrease either the AnxA1-stimulated rise in [Ca2+]i or secretion. We conclude that in health, AnxA1 contributes to the mucin layer of the tear film and ocular surface homeostasis by activating the PLC signaling pathway to increase [Ca2+]i and stimulate mucin secretion and ERK1/2, PLA2, and PLD to stimulate mucin secretion from conjunctival goblet cells.https://www.frontiersin.org/articles/10.3389/fimmu.2021.618653/fullocular surfaceannexin A1secretionspecialized pro resolving mediatorsmucin |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Anne V. Lyngstadaas Anne V. Lyngstadaas Anne V. Lyngstadaas Markus V. Olsen Markus V. Olsen Markus V. Olsen Jeffrey A. Bair Robin R. Hodges Tor P. Utheim Tor P. Utheim Tor P. Utheim Charles N. Serhan Darlene A. Dartt |
spellingShingle |
Anne V. Lyngstadaas Anne V. Lyngstadaas Anne V. Lyngstadaas Markus V. Olsen Markus V. Olsen Markus V. Olsen Jeffrey A. Bair Robin R. Hodges Tor P. Utheim Tor P. Utheim Tor P. Utheim Charles N. Serhan Darlene A. Dartt Pro-Resolving Mediator Annexin A1 Regulates Intracellular Ca2+ and Mucin Secretion in Cultured Goblet Cells Suggesting a New Use in Inflammatory Conjunctival Diseases Frontiers in Immunology ocular surface annexin A1 secretion specialized pro resolving mediators mucin |
author_facet |
Anne V. Lyngstadaas Anne V. Lyngstadaas Anne V. Lyngstadaas Markus V. Olsen Markus V. Olsen Markus V. Olsen Jeffrey A. Bair Robin R. Hodges Tor P. Utheim Tor P. Utheim Tor P. Utheim Charles N. Serhan Darlene A. Dartt |
author_sort |
Anne V. Lyngstadaas |
title |
Pro-Resolving Mediator Annexin A1 Regulates Intracellular Ca2+ and Mucin Secretion in Cultured Goblet Cells Suggesting a New Use in Inflammatory Conjunctival Diseases |
title_short |
Pro-Resolving Mediator Annexin A1 Regulates Intracellular Ca2+ and Mucin Secretion in Cultured Goblet Cells Suggesting a New Use in Inflammatory Conjunctival Diseases |
title_full |
Pro-Resolving Mediator Annexin A1 Regulates Intracellular Ca2+ and Mucin Secretion in Cultured Goblet Cells Suggesting a New Use in Inflammatory Conjunctival Diseases |
title_fullStr |
Pro-Resolving Mediator Annexin A1 Regulates Intracellular Ca2+ and Mucin Secretion in Cultured Goblet Cells Suggesting a New Use in Inflammatory Conjunctival Diseases |
title_full_unstemmed |
Pro-Resolving Mediator Annexin A1 Regulates Intracellular Ca2+ and Mucin Secretion in Cultured Goblet Cells Suggesting a New Use in Inflammatory Conjunctival Diseases |
title_sort |
pro-resolving mediator annexin a1 regulates intracellular ca2+ and mucin secretion in cultured goblet cells suggesting a new use in inflammatory conjunctival diseases |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Immunology |
issn |
1664-3224 |
publishDate |
2021-04-01 |
description |
The amount of mucin secreted by conjunctival goblet cells is regulated to ensure the optimal level for protection of the ocular surface. Under physiological conditions lipid specialized pro-resolving mediators (SPM) are essential for maintaining tissue homeostasis including the conjunctiva. The protein Annexin A1 (AnxA1) can act as an SPM. We used cultured rat conjunctival goblet cells to determine if AnxA1 stimulates an increase in intracellular [Ca2+] ([Ca2+]i) and mucin secretion and to identify the signaling pathways. The increase in [Ca2+]i was determined using fura2/AM and mucin secretion was measured using an enzyme-linked lectin assay. AnxA1 stimulated an increase in [Ca2+]i and mucin secretion that was blocked by the cell-permeant Ca2+ chelator BAPTA/AM and the ALX/FPR2 receptor inhibitor BOC2. AnxA1 increased [Ca2+]i to a similar extent as the SPMs lipoxin A4 and Resolvin (Rv) D1 and histamine. The AnxA1 increase in [Ca2+]i and mucin secretion were inhibited by blocking the phospholipase C (PLC) pathway including PLC, the IP3 receptor, the Ca2+/ATPase that causes the intracellular Ca2+ stores to empty, and blockade of Ca2+ influx. Inhibition of protein kinase C (PKC) and Ca2+/calmodulin-dependent protein kinase also decreased the AnxA1-stimulated increase in [Ca2+]i and mucin secretion. In contrast inhibitors of ERK 1/2, phospholipase A2 (PLA2), and phospholipase D (PLD) did not alter AnxA1-stimulated increase in [Ca2+]i, but did inhibit mucin secretion. Activation of protein kinase A did not decrease either the AnxA1-stimulated rise in [Ca2+]i or secretion. We conclude that in health, AnxA1 contributes to the mucin layer of the tear film and ocular surface homeostasis by activating the PLC signaling pathway to increase [Ca2+]i and stimulate mucin secretion and ERK1/2, PLA2, and PLD to stimulate mucin secretion from conjunctival goblet cells. |
topic |
ocular surface annexin A1 secretion specialized pro resolving mediators mucin |
url |
https://www.frontiersin.org/articles/10.3389/fimmu.2021.618653/full |
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