Regulation of Conjunctival Goblet Cell Secretion by Ca2+and Protein Kinase C

Dartt, Darlene A.; Rios, Jose R; Kanno, Hiroaki; Rawe, Ian; Zieske, James D.; Ralda, Nereida; Hodges, Robin R.; Zoukhri, Driss

doi:10.1006/exer.2000.0915

Cited by 36 publications

(28 citation statements)

References 29 publications

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“…We also showed that cholinergic agonists activated PKC (Dartt, et al, 2000). However, we were unable to determine a direct role of PKC in mucin secretion as the inhibitors themselves increased secretion.…”

Section: Effect Of Pkc Inhibitors On Cholinergic Agonist-stimulated Pmentioning

confidence: 65%

“…As we were previously unable to determine the role of PKC in HMWGC secretion due to an increase in basal secretion by PKC inhibitors themselves (Dartt, et al, 2000), we determined the effects of PKC inhibitors on p42/p44 MAPK activation. The PKC inhibitors calphostin C and chelethyrine chloride did not affect basal MAPK activity but inhibited cholinergic-agonist stimulated p42/p44 MAPK.…”

Section: Discussionmentioning

confidence: 99%

“…We previously showed that the phorbol esters PMA and PdBu increased secretion of HMWGC from conjunctival pieces (Dartt, et al, 2000). In addition, we have shown that activation of p42/p44 MAPK also leads to HMWGC secretion (Kanno, et al, 2003).…”

Section: Effect Of Time and Concentration Of Pma On P42/p44 Mapk Actimentioning

confidence: 92%

“…We previously showed that increasing intracellular Ca 2+ with the calcium ionophore ionomycin stimulated mucin secretion from conjunctival pieces in a time-and concentrationdependent manner (Dartt, et al, 2000). To determine if Ca 2+ -stimulated secretion was dependent upon activation of the EGF receptor, conjunctival pieces were pretreated with AG1478 (10 −7 M) for 10 min before incubation with ionomycin (10 −6 M) for 10 min.…”

Section: Effect Of Inhibition Of the Egf Receptor On Ca 2+ -Stimulatementioning

confidence: 99%

“…Ectodomain shedding of EGF has been shown to occur in the lacrimal gland (Chen, et al, 2006) while shedding of TGFα and HB-EGF occurs in many other tissues (Prenzel, et al, 2000). Determining if ectodomain shedding occurs in conjunctival goblet cells in response to cholinergic stimulation and the identification of the specific growth factor shed is under investigation.As we were previously unable to determine the role of PKC in HMWGC secretion due to an increase in basal secretion by PKC inhibitors themselves (Dartt, et al, 2000), we determined the effects of PKC inhibitors on p42/p44 MAPK activation. The PKC inhibitors calphostin C and chelethyrine chloride did not affect basal MAPK activity but inhibited cholinergic-agonist stimulated p42/p44 MAPK.…”

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confidence: 99%

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Effect of protein kinase C and Ca2+ on p42/p44 MAPK, Pyk2, and Src activation in rat conjunctival goblet cells

Hodges

Horikawa

Ríos

et al. 2007

Experimental Eye Research

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Conjunctival goblet cells synthesize and secrete mucins onto the ocular surface to lubricate it and protect it from bacterial infections. Mucin secretion is under neural control, and cholinergic agonists released from parasympathetic nerves are major stimuli of this secretion. The signal transduction pathways these agonists use to stimulate secretion involve activating protein kinase C (PKC) and increasing intracellular [Ca 2+ ] to activate the non-receptor kinases Pyk2 and p60Src (Src) to transactivate the EGF receptor. Transactivation of the EGF receptor activates a kinase cascade culminating in the activation of p42/p44 MAPK (MAPK) and ultimately that leads to secretion of high molecular weight glycocongujates (HMWGC), including mucins. To further examine the roles of PKC and Ca 2+ in the activation of MAPK, Pyk2, and Src in mucin secretion, rat conjunctival pieces and cultured goblet cells were incubated with the PKC activator phorbol myristate acid (PMA), the cholinergic agonist carbachol, or the calcium ionophore, ionomycin for varying times. Conjunctival pieces were preincubated with PKC inhibitors 10 mins prior to addition of carbachol (10 −4 M) for 10 min. The amount of phosphorylated (activated) MAPK, Pyk2 and Src was determined by western blotting techniques using antibodies specific to the phosphorylated forms of each kinase. PMA significantly increased the activation of MAPK, Pyk2, and Src in a time and concentrationdependent manner. PMA-stimulated MAPK activity was completely inhibited by the EGF receptor inhibitor AG1478 (10 −7 M). Carbachol-stimulated MAPK activity was inhibited by three PKC inhibitors, calphostin C, chelethyrine, and staurosporine. Ionomycin (10 −6 M)-stimulated MAPK activity was inhibited 66% by AG1478 (10 −7 M). Ionomycin also significantly increased Pyk2 and Src in time dependent manner. PKC and ionomycin also activated p42/p44 MAPL, Pyk2, and Src in cultured conjunctival goblet cells. We conclude that PKC and intracellular Ca 2+ activate Pyk2 and Src and phosphorylated the EGF receptor leading to stimulation of MAPK in conjunctival goblet cells. Keywords goblet cells; signal transduction; MAPK; mucin secretionGoblet cells of the conjunctiva are responsible for synthesis, storage, and secretion of mucins, which make up the mucous layer of the tear film (Dartt, 2004, Gipson andArgueso, 2003). Mucins serve to lubricate the ocular surface, protect from bacterial infections and provide for a smooth refractive surface. These cells are highly specialized epithelial cells that are * author to whom correspondence should be addressed: 20 Staniford Street, Boston, MA 02114. Tel: 617-912-7424; FAX: 617-912-0104; email: robin.hodges@schepens.harvard.edu. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form...

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Section: Effect Of Pkc Inhibitors On Cholinergic Agonist-stimulated Pmentioning

confidence: 65%

Section: Discussionmentioning

confidence: 99%

Section: Effect Of Time and Concentration Of Pma On P42/p44 Mapk Actimentioning

confidence: 92%

Section: Effect Of Inhibition Of the Egf Receptor On Ca 2+ -Stimulatementioning

confidence: 99%

mentioning

confidence: 99%

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Effect of protein kinase C and Ca2+ on p42/p44 MAPK, Pyk2, and Src activation in rat conjunctival goblet cells

Hodges

Horikawa

Ríos

et al. 2007

Experimental Eye Research

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Maresin 1, a specialized proresolving mediator, stimulates intracellular [Ca²⁺] and secretion in conjunctival goblet cells

Olsen

Lyngstadaas

Bair

et al. 2020

Journal Cellular Physiology

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Calcium Signalling in Ocular Tissues: Functional Activity of G-protein and Tyrosine–Kinase Coupled Receptors

Duncan¹,

Collison²

2002

Experimental Eye Research

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Regulation of Conjunctival Goblet Cell Secretion by Ca2+and Protein Kinase C

Cited by 36 publications

References 29 publications

Effect of protein kinase C and Ca2+ on p42/p44 MAPK, Pyk2, and Src activation in rat conjunctival goblet cells

Effect of protein kinase C and Ca2+ on p42/p44 MAPK, Pyk2, and Src activation in rat conjunctival goblet cells

Maresin 1, a specialized proresolving mediator, stimulates intracellular [Ca²⁺] and secretion in conjunctival goblet cells

Calcium Signalling in Ocular Tissues: Functional Activity of G-protein and Tyrosine–Kinase Coupled Receptors

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Regulation of Conjunctival Goblet Cell Secretion by Ca2+and Protein Kinase C

Cited by 36 publications

References 29 publications

Effect of protein kinase C and Ca2+ on p42/p44 MAPK, Pyk2, and Src activation in rat conjunctival goblet cells

Effect of protein kinase C and Ca2+ on p42/p44 MAPK, Pyk2, and Src activation in rat conjunctival goblet cells

Maresin 1, a specialized proresolving mediator, stimulates intracellular [Ca2+] and secretion in conjunctival goblet cells

Calcium Signalling in Ocular Tissues: Functional Activity of G-protein and Tyrosine–Kinase Coupled Receptors

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Product

Resources

About

Maresin 1, a specialized proresolving mediator, stimulates intracellular [Ca²⁺] and secretion in conjunctival goblet cells