mast cell tryptase controls paracellular permeability of the intestine

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Mast Cell Tryptase Controls Paracellular Permeability of the Intestine. 生科四甲 陳子甯. 作者假設:. mast cells signal to colonocytes by release of tryptase and activation of PAR2, and that PAR2 couples to ERK1/2 by a β ARR-dependent mechanism to regulate TJs. TJ: tight junction. - PowerPoint PPT Presentation

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  • Mast Cell Tryptase Controls Paracellular Permeabilityof the Intestine

  • mast cells signal to colonocytes by release of tryptase and activation of PAR2, and that PAR2 couples to ERK1/2 by a ARR-dependent mechanism to regulate TJs.

    ERK1/2extracellular signal-regulated kinases 1/2ARR:-arrestinTJ: tight junction

  • PAR2TJs

    mast cell tryptasePAR2colonocytes

    .ERK1/2ARRPAR2

    ERK1/2extracellular signal-regulated kinases 1/2ARR:-arrestin

  • Mast cell:

  • Model for -arrestin-dependent activation of ERK1/2 by PAR2

    microtubule-associated proteins (MAPs) phospholipase A2 (PLA2) Ca2+

  • FIG. 1. Expression of mRNA encoding PAR2 and trypsinogen IV by colonocytes.

    525bp400bp525bp400bpcpcTcpcTNCM460T84PAR2trypsinogenIVNCM460T84

  • FIG. 2. PAR2-mediated mobilization of [Ca2+]i in colonocytes.

    AP:AA-SLIGKV RP:APpeptide-VKGILScontrol

  • FIG. 3. PAR2-mediated mobilization of [Ca2+]i in colonocytes.

    trypsinPAR2 trypsinPAR2c

  • FIG. 4. Characterization of CHMC.

    mast cellIgE bindingdegranuledCHMC

    CHMCmast cellSBTI: Soybean trypsin inhibitormast celltryptasedegranule

  • Flow Cytometry

  • FIG. 5. Mast cell signaling to colonocytes.

    48/80mast cell degranule mast celldegranuletryptasesupernatant formcolonocytes

  • TER ( transepithelial resistance ) & paracellular permeability

  • FIG. 6. Effects of PAR2 agonists on paracellular permeability.

    PAR2 Activation Increases Paracellular Permeability

    HRP

  • ZO-1E-cadhesinApicalBasalZo-1E-cadhesin

  • FIG. 7. PAR2-induced redistribution of TJ proteins and F-actin detected by microscopy

    ZO-1Claudin-1F-actinoccludinIsotype

    PAR2TJ protein F-actin redistribution

  • FIG. 8. PAR2-induced redistribution of TJ proteins detected by subcellular fractionation and Western blotting.

    PAR2TJ proteinsredistributionE-cadherin is a transmembrane glycoprotein that mediates epithelial cell-to-cell adhesion.

  • FIG. 9. Effects of mast cell products on paracellular permeability.

    PAR2paracellular permeability

  • FIG. 10. ERK1/2-mediated changes in paracellular permeability.

    ERK1/2 paracellular permeabilityPAR2ERK1/2PAR2paracellular permeabilityUO126ERK1/2

  • FIG. 11. Down-regulation of ARR using siRNAARR1ARR2498bp381bpARR cytoplasm-nucleus messengerPAR2ERK1/2 ARR ERK1/2

  • FIG. 12. ARR-mediated changes in paracellular permeability.

    Oligofectamine ARR ERK1/2 bindingPAR2ARR ERK1/2colonocytes

  • Conclusion

    Colonocytes expressed PAR2 mRNA and responded to PAR2 agonists with increased [Ca2+]i

    Mast cells signal to colonocytes in a paracrine manner by release of tryptase and activation of PAR2.

    ARRERK1/2colonocytes permeability

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