{"id":{"repo_id":"east-anglia","oai_identifier":"oai:ueaeprints.uea.ac.uk:53377"},"canonical_url":"https://search.dev.ndltd.org/etd/east-anglia/oai:ueaeprints.uea.ac.uk:53377","repository":{"repo_id":"east-anglia","name":"University of East Anglia","base_url":"https://ueaeprints.uea.ac.uk/cgi/oai2"},"display":{"title":"Role of matriptase in the regulation of epithelial barrier permeability studied using MDCK cells","abstract":"The!typeNII!transmembrane!serine!protease!matriptase!plays!an!important!role!in! the!integrity!of!epithelial!barriers.!However,!the!molecular!mechanisms!underlying! the!role!of!matriptase!are!unknown.! To! study! these!mechanisms,! two!variants!of!MadinNDarby!canine!kidney!(MDCK)! cells!were! used,! together!with! the! “calciumNswitch”!model! of! epithelial! function! with!measurements!of!transepithelial!electrical!resistance!(TEER).! Inhibitors!of!matriptase!proteolytic!activity!delayed!the!restoration!of!TEER!after! calciumNswitch! in!MDCKNI,! which! develop! high! TEER! and! lack! the! “leaky”! tight! junction!protein!claudinN2,!but!not!MDCKNII.!This!effect!was!confirmed!in!MDCKNI,! established!to!stably!express!matriptase!targeted!shRNA.! The!influence!of!matriptase!inhibition!on!MDCKNI!was!shown!not!to!involve!altered! expression! or! assembly! of! relevant! components! of! paracelluar! junctions! or! cytoskeleton.! This! excluded! a! role! for! claudinN2! in! the! function! of! matriptase,! which!had!previously!been!shown!in!human!CacoN2!cells,!and!this!was!confirmed! using!MDCKNI!cells!stably!overexpressing!claudinN2.! To! investigate! the! claudinN2Nindependent! function! of! matriptase,! a! candidate! substrate! approach! was! used.! Proteolytic! activation! of! proNHGF/cNMet,! PARN2,! ENaC,!EGFR!and!prostasin!by!matriptase!has!effects!on!epithelial!cell!function,!but! none!were! found! to!have!a! role! in!matriptase! restoration!of!TEER!after! calciumN switch!in!MDCKNI!cells.!As!the!direct!proteolytic!target!of!matriptase!could!not!be! identified,! potential! mediators! were! studied! using! arrays! for! phosphorylated! signalling!proteins!and!inflammatory!cytokines.!ILN1β!and!complement!component! C5!were! identified!as!genes!downregulated!by!matriptase! inhibition,!while! ILN13! showed!upregulation.! This!work!has!confirmed!the!key!role!of!matriptase!activity!in!regulating!epithelial! barrier!integrity.!The!differential!properties!of!MDCKNI!and!MDCKNII!cells!excluded! a! role! for! claudinN2.! None! of! the! known! proteolytic! targets! of! matriptase! were! involved,!however,!changes!in!cytokine!gene!expression!may!be!a!potential!route! for!matriptase!effects!on!epithelial!barrier!maintenance!","abstract_html":"The!typeNII!transmembrane!serine!protease!matriptase!plays!an!important!role!in! the!integrity!of!epithelial!barriers.!However,!the!molecular!mechanisms!underlying! the!role!of!matriptase!are!unknown.! To! study! these!mechanisms,! two!variants!of!MadinNDarby!canine!kidney!(MDCK)! cells!were! used,! together!with! the! “calciumNswitch”!model! of! epithelial! function! with!measurements!of!transepithelial!electrical!resistance!(TEER).! Inhibitors!of!matriptase!proteolytic!activity!delayed!the!restoration!of!TEER!after! calciumNswitch! in!MDCKNI,! which! develop! high! TEER! and! lack! the! “leaky”! tight! junction!protein!claudinN2,!but!not!MDCKNII.!This!effect!was!confirmed!in!MDCKNI,! established!to!stably!express!matriptase!targeted!shRNA.! The!influence!of!matriptase!inhibition!on!MDCKNI!was!shown!not!to!involve!altered! expression! or! assembly! of! relevant! components! of! paracelluar! junctions! or! cytoskeleton.! This! excluded! a! role! for! claudinN2! in! the! function! of! matriptase,! which!had!previously!been!shown!in!human!CacoN2!cells,!and!this!was!confirmed! using!MDCKNI!cells!stably!overexpressing!claudinN2.! To! investigate! the! claudinN2Nindependent! function! of! matriptase,! a! candidate! substrate! approach! was! used.! Proteolytic! activation! of! proNHGF/cNMet,! PARN2,! ENaC,!EGFR!and!prostasin!by!matriptase!has!effects!on!epithelial!cell!function,!but! none!were! found! to!have!a! role! in!matriptase! restoration!of!TEER!after! calciumN switch!in!MDCKNI!cells.!As!the!direct!proteolytic!target!of!matriptase!could!not!be! identified,! potential! mediators! were! studied! using! arrays! for! phosphorylated! signalling!proteins!and!inflammatory!cytokines.!ILN1β!and!complement!component! C5!were! identified!as!genes!downregulated!by!matriptase! inhibition,!while! ILN13! showed!upregulation.! This!work!has!confirmed!the!key!role!of!matriptase!activity!in!regulating!epithelial! barrier!integrity.!The!differential!properties!of!MDCKNI!and!MDCKNII!cells!excluded! a! role! for! claudinN2.! None! of! the! known! proteolytic! targets! of! matriptase! were! involved,!however,!changes!in!cytokine!gene!expression!may!be!a!potential!route! for!matriptase!effects!on!epithelial!barrier!maintenance!","abstract_has_math":false,"creators":["Elghadban, Salma"],"institution":"University of East Anglia","degree_name":"phd","degree_level":"doctoral","degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2014,"date_issued":"2014-09","date_published":"2014-09","updated_at":"2026-07-24T02:12:08Z","subjects":[],"languages":["en"],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":null,"outbound_label":null,"outbound_source":null},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:creator","label":"Author","values":["Elghadban, Salma"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2014-09"]},{"key":"dc:date.issued","label":"Date","values":["2014-09"]},{"key":"dc:publisher.department","label":"Dc Publisher Department","values":["School of Biological Sciences"]},{"key":"dc:publisher.institution","label":"Dc Publisher Institution","values":["University of East Anglia"]},{"key":"dc:relation.isreferencedby","label":"Dc Relation Isreferencedby","values":["https://ueaeprints.uea.ac.uk/id/eprint/53377/"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]},{"key":"dc:type.qualificationlevel","label":"Dc Type Qualificationlevel","values":["doctoral"]},{"key":"dc:type.qualificationname","label":"Dc Type Qualificationname","values":["phd"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://ueaeprints.uea.ac.uk/id/eprint/53377/1/2014ElghadbanSPhD.pdf"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["The!typeNII!transmembrane!serine!protease!matriptase!plays!an!important!role!in! the!integrity!of!epithelial!barriers.!However,!the!molecular!mechanisms!underlying! the!role!of!matriptase!are!unknown.! To! study! these!mechanisms,! two!variants!of!MadinNDarby!canine!kidney!(MDCK)! cells!were! used,! together!with! the! “calciumNswitch”!model! of! epithelial! function! with!measurements!of!transepithelial!electrical!resistance!(TEER).! Inhibitors!of!matriptase!proteolytic!activity!delayed!the!restoration!of!TEER!after! calciumNswitch! in!MDCKNI,! which! develop! high! TEER! and! lack! the! “leaky”! tight! junction!protein!claudinN2,!but!not!MDCKNII.!This!effect!was!confirmed!in!MDCKNI,! established!to!stably!express!matriptase!targeted!shRNA.! The!influence!of!matriptase!inhibition!on!MDCKNI!was!shown!not!to!involve!altered! expression! or! assembly! of! relevant! components! of! paracelluar! junctions! or! cytoskeleton.! This! excluded! a! role! for! claudinN2! in! the! function! of! matriptase,! which!had!previously!been!shown!in!human!CacoN2!cells,!and!this!was!confirmed! using!MDCKNI!cells!stably!overexpressing!claudinN2.! To! investigate! the! claudinN2Nindependent! function! of! matriptase,! a! candidate! substrate! approach! was! used.! Proteolytic! activation! of! proNHGF/cNMet,! PARN2,! ENaC,!EGFR!and!prostasin!by!matriptase!has!effects!on!epithelial!cell!function,!but! none!were! found! to!have!a! role! in!matriptase! restoration!of!TEER!after! calciumN switch!in!MDCKNI!cells.!As!the!direct!proteolytic!target!of!matriptase!could!not!be! identified,! potential! mediators! were! studied! using! arrays! for! phosphorylated! signalling!proteins!and!inflammatory!cytokines.!ILN1β!and!complement!component! C5!were! identified!as!genes!downregulated!by!matriptase! inhibition,!while! ILN13! showed!upregulation.! This!work!has!confirmed!the!key!role!of!matriptase!activity!in!regulating!epithelial! barrier!integrity.!The!differential!properties!of!MDCKNI!and!MDCKNII!cells!excluded! a! role! for! claudinN2.! None! of! the! known! proteolytic! targets! of! matriptase! were! involved,!however,!changes!in!cytokine!gene!expression!may!be!a!potential!route! for!matriptase!effects!on!epithelial!barrier!maintenance!"]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Role of matriptase in the regulation of epithelial barrier permeability studied using MDCK cells"]}]}],"canonical_facts":{"dc:creator":["Elghadban, Salma"],"dc:date":["2014-09"],"dc:date.issued":["2014-09"],"dc:description.abstract":["The!typeNII!transmembrane!serine!protease!matriptase!plays!an!important!role!in! the!integrity!of!epithelial!barriers.!However,!the!molecular!mechanisms!underlying! the!role!of!matriptase!are!unknown.! To! study! these!mechanisms,! two!variants!of!MadinNDarby!canine!kidney!(MDCK)! cells!were! used,! together!with! the! “calciumNswitch”!model! of! epithelial! function! with!measurements!of!transepithelial!electrical!resistance!(TEER).! Inhibitors!of!matriptase!proteolytic!activity!delayed!the!restoration!of!TEER!after! calciumNswitch! in!MDCKNI,! which! develop! high! TEER! and! lack! the! “leaky”! tight! junction!protein!claudinN2,!but!not!MDCKNII.!This!effect!was!confirmed!in!MDCKNI,! established!to!stably!express!matriptase!targeted!shRNA.! The!influence!of!matriptase!inhibition!on!MDCKNI!was!shown!not!to!involve!altered! expression! or! assembly! of! relevant! components! of! paracelluar! junctions! or! cytoskeleton.! This! excluded! a! role! for! claudinN2! in! the! function! of! matriptase,! which!had!previously!been!shown!in!human!CacoN2!cells,!and!this!was!confirmed! using!MDCKNI!cells!stably!overexpressing!claudinN2.! To! investigate! the! claudinN2Nindependent! function! of! matriptase,! a! candidate! substrate! approach! was! used.! Proteolytic! activation! of! proNHGF/cNMet,! PARN2,! ENaC,!EGFR!and!prostasin!by!matriptase!has!effects!on!epithelial!cell!function,!but! none!were! found! to!have!a! role! in!matriptase! restoration!of!TEER!after! calciumN switch!in!MDCKNI!cells.!As!the!direct!proteolytic!target!of!matriptase!could!not!be! identified,! potential! mediators! were! studied! using! arrays! for! phosphorylated! signalling!proteins!and!inflammatory!cytokines.!ILN1β!and!complement!component! C5!were! identified!as!genes!downregulated!by!matriptase! inhibition,!while! ILN13! showed!upregulation.! This!work!has!confirmed!the!key!role!of!matriptase!activity!in!regulating!epithelial! barrier!integrity.!The!differential!properties!of!MDCKNI!and!MDCKNII!cells!excluded! a! role! for! claudinN2.! None! of! the! known! proteolytic! targets! of! matriptase! were! involved,!however,!changes!in!cytokine!gene!expression!may!be!a!potential!route! for!matriptase!effects!on!epithelial!barrier!maintenance!"],"dc:format":["application/pdf"],"dc:identifier.uri":["https://ueaeprints.uea.ac.uk/id/eprint/53377/1/2014ElghadbanSPhD.pdf"],"dc:language":["en"],"dc:publisher.department":["School of Biological Sciences"],"dc:publisher.institution":["University of East Anglia"],"dc:relation.isreferencedby":["https://ueaeprints.uea.ac.uk/id/eprint/53377/"],"dc:title":["Role of matriptase in the regulation of epithelial barrier permeability studied using MDCK cells"],"dc:type":["Thesis"],"dc:type.qualificationlevel":["doctoral"],"dc:type.qualificationname":["phd"]},"updated_at":"2026-07-24T02:12:08Z"}