In vivo and in vitro formation of the junctional complex in choroid epithelium. A freeze-etching study

The junctional complex of choroid epithelial cells was studied during in vivo formation, disaggregation after trypsin treatment, and in vitro reaggregation. The in vivo formation begins with the occurrence of amorphous patches of particles followed by the formation of small particulate rows and polygonal-ordered particle assemblies. Further arrangement of the zonula occludens continues with the confluence of particles and smooth contoured ridges. At the 9th day stage a fully developed zonula occludens has developed. In a subsequent step nexus become integrated within the tight junction formation. Disaggregation after trypsination results in fragmentation of the zonulae occludentes. Parts of the disassembling aggregates become incorporated in vacuoles indicating an endocytotic mode of "digestion". The in vitro reconstruction of the zonula occludens proceeds from remnants of the former zonula occludens. On the 3rd to 4th day of cultivation mature tight junctions are visible. In vitro integrations of nexus were observed during a later phase. On the 7th day, cultivated choroid epithelial cells reveal well differentiated junctional complexes consisting of continuous zonulae occludentes and integrated gap junctions.