Assembly and sealing of tight junctions: possible participation of G-proteins, phospholipase C, protein kinase C and calmodulin

J Membr Biol. 1991 Jun;122(3):193-202. doi: 10.1007/BF01871420.


The making and sealing of a tight junction (TJ) requires cell-cell contacts and Ca2+, and can be gauged through the development of transepithelial electrical resistance (TER) and the accumulation of ZO-1 peptide at the cell borders. We observe that pertussis toxin increases TER, while AIF3 and carbamil choline (carbachol) inhibit it, and 5-guanylylimidodiphosphate (GTPTs) blocks the development of a cell border pattern of ZO-1, suggesting that G-proteins are involved. Phospholipase C (PLC) and protein kinase C (PKC) probably participate in these processes since (i) activation of PLC by thyrotropin-1 releasing hormone increases TER, and its inhibition by neomycin blocks the development of this resistance; (ii) 1,2-dioctanoylglycerol, an activator of PKC, stimulates TER development, while polymyxin B and 1-(5-isoquinoline sulfonyl)-2-methyl-piperazine dihydrochloride (H7), which inhibit this enzyme, abolish TER. Addition of 3-isobutyl-1-methyl-xanthine, dB-cAMP or forskolin do not enhance the value of TER, but have just the opposite effect. Trifluoperazine and calmidazoline inhibit TER development, suggesting that calmodulin (CaM) also plays a role in junction formation. These results indicate that junction formation may be controlled by a network of reactions where G-proteins, phospholipase C, adenylate cyclase, protein kinase C and CaM are involved.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
  • Adenylate Cyclase Toxin
  • Animals
  • Calcium / physiology
  • Calmodulin / physiology*
  • Carbachol / pharmacology
  • Cells, Cultured
  • Electric Conductivity / drug effects
  • Electric Conductivity / physiology
  • Epithelial Cells
  • Epithelium / metabolism
  • Epithelium / physiology
  • GTP-Binding Proteins / physiology*
  • Intercellular Junctions / metabolism
  • Intercellular Junctions / physiology*
  • Isoquinolines / pharmacology
  • Kidney / cytology
  • Kidney / metabolism
  • Kidney / physiology
  • Membrane Potentials / drug effects
  • Membrane Potentials / physiology
  • Neomycin / pharmacology
  • Pertussis Toxin
  • Piperazines / pharmacology
  • Polymyxin B / pharmacology
  • Protein Kinase C / antagonists & inhibitors
  • Protein Kinase C / physiology*
  • Trifluoperazine / pharmacology
  • Type C Phospholipases / physiology*
  • Virulence Factors, Bordetella / pharmacology


  • Adenylate Cyclase Toxin
  • Calmodulin
  • Isoquinolines
  • Piperazines
  • Virulence Factors, Bordetella
  • Trifluoperazine
  • 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
  • Carbachol
  • Pertussis Toxin
  • Protein Kinase C
  • Type C Phospholipases
  • GTP-Binding Proteins
  • Neomycin
  • Polymyxin B
  • Calcium