Serine proteases decrease intestinal epithelial ion permeability by activation of protein kinase Cζ

Swystun, Veronica A.; Renaux, Bernard; Wen, Shoubin; Peplowski, Michael A.; Hollenberg, Morley D.; MacNaughton, Wallace K.

doi:10.1152/ajpgi.00096.2009

Cited by 29 publications

(37 citation statements)

References 61 publications

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“…Furthermore, this result was not due to a lack of sensitivity of the Ussing chambers because changes in permeability could be detected when present, such as after infection or EGTA addition (10,34). Recent data point to differences in epithelial barrier modulation by the PAR-2-activating peptide and endogenous agonists with enzymatic activity (39). Therefore, we tested whether trypsin, as an endogenous activator of PAR-2, was capable of modulating intestinal permeability.…”

Section: Discussionmentioning

confidence: 96%

PAR-2-mediated control of barrier function and motility differs between early and late phases of postinfectious gut dysfunction in the rat

Fernandez-Blanco¹,

Hollenberg

Martínez

et al. 2013

American Journal of Physiology-Gastrointestinal and Liver Physi

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Fernández-Blanco JA, Hollenberg MD, Martínez V, Vergara P. PAR-2-mediated control of barrier function and motility differs between early and late phases of postinfectious gut dysfunction in the rat. Am J Physiol Gastrointest Liver Physiol 304: G390 -G400, 2013. First published December 13, 2012; doi:10.1152/ajpgi.00387.2012.-Proteinase-activated receptor-2 (PAR-2) and mast cell (MC) mediators contribute to inflammatory and functional gastrointestinal disorders. We aimed to characterize jejunal PAR-2-mediated responses and the potential MC involvement in the early and late phases of a rat model of postinfectious gut dysfunction. Jejunal tissues of control and Trichinella spiralis-infected (14 and 30 days postinfection) rats, treated or not with the MC stabilizer, ketotifen, were used. Histopathology and immunostaining were used to characterize inflammation, PAR-2 expression, and mucosal and connective tissue MCs. Epithelial barrier function (hydroelectrolytic transport and permeability) and motility were assessed in vitro in basal conditions and after PAR-2 activation. Intestinal inflammation on day 14 postinfection (early phase) was significantly resolved by day 30 (late phase) although MC counts and epithelial permeability remained increased. PAR-2-mediated ion transport (Ussing chambers, in vitro) and epithelial surface PAR-2 expression were reduced in the early phase, with a trend toward normalization during the late phase. In control conditions, PAR-2 activation (organ bath) induced biphasic motor responses (relaxation followed by excitation). At 14 days postinfection, spontaneous contractility and PAR-2-mediated relaxations were enhanced; motor responses were normalized on day 30. Postinfectious changes in PAR-2 functions were not affected by ketotifen treatment. We concluded that, in the rat model of Trichinella spiralis infection, alterations of intestinal PAR-2 function and expression depend on the inflammatory phase considered. A lack of a ketotifen effect suggests no interplay between MCs and PAR-2-mediated motility and ion transport alterations. These observations question the role of MC mediators in PAR-2-modulating postinfectious gut dysfunction.

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Section: Discussionmentioning

confidence: 96%

PAR-2-mediated control of barrier function and motility differs between early and late phases of postinfectious gut dysfunction in the rat

Fernandez-Blanco¹,

Hollenberg

Martínez

et al. 2013

American Journal of Physiology-Gastrointestinal and Liver Physi

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“…In general, low levels of trypsin and trypsin-like proteases such as cap1 and matriptase (cap3) are potent inducers of TJ formation in a variety of epithelia (25,(46)(47)(48). The constitutive cap1 KO mice died within 60 d after birth due to impaired skin barrier function and fatal dehydration (49).…”

Section: Discussionmentioning

confidence: 99%

The Cap1–claudin-4 regulatory pathway is important for renal chloride reabsorption and blood pressure regulation

Gong

Yang³

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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The paracellular pathway through the tight junction provides an important route for transepithelial chloride reabsorption in the kidney, which regulates extracellular salt content and blood pressure. Defects in paracellular chloride reabsorption may in theory cause deregulation of blood pressure. However, there is no evidence to prove this theory or to demonstrate the in vivo role of the paracellular pathway in renal chloride handling. Here, using a tissue-specific KO approach, we have revealed a chloride transport pathway in the kidney that requires the tight junction molecule claudin-4. The collecting duct-specific claudin-4 KO animals developed hypotension, hypochloremia, and metabolic alkalosis due to profound renal wasting of chloride. The claudin-4-mediated chloride conductance can be regulated endogenously by a proteasechannel-activating protease 1 (cap1). Mechanistically, cap1 regulates claudin-4 intercellular interaction and membrane stability. A putative cap1 cleavage site has been identified in the second extracellular loop of claudin-4, mutation of which abolished its regulation by cap1. The cap1 effects on paracellular chloride permeation can be extended to other proteases such as trypsin, suggesting a general mechanism may also exist for proteases to regulate the tight junction permeabilities. Together, we have discovered a theory that paracellular chloride permeability is physiologically regulated and essential to renal salt homeostasis and blood pressure control.chloride channel | epithelium | hypertension C hloride is the most abundant extracellular anion and thereby determines extracellular fluid volume (ECFV) and blood pressure (BP) (1, 2). The kidney plays a vital role in ECFV and BP control through complex regulatory mechanisms acting upon ion channels and transporters located in the aldosterone-sensitive distal nephron (ASDN) (3, 4). The ASDN comprises the distal convoluted tubule (DCT), the connecting tubule (CNT), and the collecting duct (CD). The primary chloride transport mechanism in the DCT is through the thiazide-sensitive Na + /Cl − cotransporter (NCC) to reabsorb Cl − coupled with equal moles of Na + (5). The chloride transport mechanism in the CNT and CD, on the other hand, has been under debate for many years. Recent advances have identified an electroneutral transport pathway for chloride using the Cl − /bicarbonate exchanger (Slc26a4; pendrin) (6) and the Na + -driven Cl − /bicarbonate exchanger (Slc4a8; NDCBE) (7) in the β-intercalated cells (ICs) of CNT and CD. However, such an electroneutral pathway is not able to couple Cl − reabsorption with epithelial sodium channel (ENaC)-based Na + reabsorption that takes place in the principal cells (PCs) of CNT and CD (8), despite many efforts to connect ENaC and pendrin gene expression through endocrine or paracrine mechanisms (9, 10). We and others have previously demonstrated the presence of a paracellular Cl − pathway or "chloride shunt" in vitro in the CNT and CD cells (11,12). The paracellular Cl − channel is made of a key claud...

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“…The pathways that regulate claudin-2 protein turnover and target it for degradation via lysosomes (16) remain to be established; however, our data implicate a matriptase-mediated proteolytic pathway that facilitates the turnover of claudin-2 during AJC assembly that is associated with PKCζ activation. Interestingly, PKCζ signaling has been implicated recently in trypsin and chymotrypsin-enhanced barrier formation (23).…”

Section: Discussionmentioning

confidence: 99%

“…Low levels of trypsin and certain other endopeptidases are potent inducers of intercellular AJC formation in intestinal epithelial monolayers (23,25), whereas higher levels of these enzymes disrupt epithelial barriers. Epithelial barrier formation may also be suppressed by serine protease inhibitors (25), further implicating serine protease activities in AJC assembly.…”

Section: Discussionmentioning

confidence: 99%

Membrane-anchored serine protease matriptase regulates epithelial barrier formation and permeability in the intestine

Buzza

Netzel‐Arnett

Shea‐Donohue³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

154

192

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The intestinal epithelium serves as a major protective barrier between the mammalian host and the external environment. Here we show that the transmembrane serine protease matriptase plays a pivotol role in the formation and integrity of the intestinal epithelial barrier. St14 hypomorphic mice, which have a 100-fold reduction in intestinal matriptase mRNA levels, display a 35% reduction in intestinal transepithelial electrical resistance (TEER). Matriptase is expressed during intestinal epithelial differentiation and colocalizes with E-cadherin to apical junctional complexes (AJC) in differentiated polarized Caco-2 monolayers. Inhibition of matriptase activity using a specific peptide inhibitor or by knockdown of matriptase by siRNA disrupts the development of TEER in barrierforming Caco-2 monolayers and increases paracellular permeability to macromolecular FITC-dextran. Loss of matriptase was associated with enhanced expression and incorporation of the permeabilityassociated, "leaky" tight junction protein claudin-2 at intercellular junctions. Knockdown of claudin-2 enhanced the development of TEER in matriptase-silenced Caco-2 monolayers, suggesting that the reduced barrier integrity was caused, at least in part, by an inability to regulate claudin-2 expression and incorporation into junctions. We find that matriptase enhances the rate of claudin-2 protein turnover, and that this is mediated indirectly through an atypical PKCζ-dependent signaling pathway. These results support a key role for matriptase in regulating intestinal epithelial barrier competence, and suggest an intriguing link between pericellular serine protease activity and tight junction assembly in polarized epithelia.claudin-2 | intestinal barrier | St14 | type II transmembrane serine protease | tight junction

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Serine proteases decrease intestinal epithelial ion permeability by activation of protein kinase Cζ

Cited by 29 publications

References 61 publications

PAR-2-mediated control of barrier function and motility differs between early and late phases of postinfectious gut dysfunction in the rat

PAR-2-mediated control of barrier function and motility differs between early and late phases of postinfectious gut dysfunction in the rat

The Cap1–claudin-4 regulatory pathway is important for renal chloride reabsorption and blood pressure regulation

Membrane-anchored serine protease matriptase regulates epithelial barrier formation and permeability in the intestine

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