AIM:To explore gap junctional intercellular communication(GJIC) in cultured human Tenon’s capsule fibroblasts(HTFs) and detect the expression and distribution of connexin43(Cx43).METHODS:GJIC were demonstrated by scrape-loading and Lucifer yellow dye transfer methods.RT-PCR and Western blot were carried out to detect mRNA and protein expression of Cx43.Immunocytochemistry was performed to observe the distribution of Cx43.RESULTS:Cultured HTFs demonstrated functional GJIC in vitro.The transporting degree of Lucifer yellow dye was more than 6 grades.The mRNA and protein expression of Cx43 were also detected.Cx43 were located on cells’ membrane by immunocytochemistry.CONCLUSION:Cultured HTFs have GJIC function which confer these cells the ability to establish and maintain intercellular communication mediated by Cx43.This property may be critical in maintaining function synchronization of fibroblasts during wound-healing process.