, Volume 162, Issue 2, pp 235-252

Freeze-fracture replication of junctional complexes in unincubated and incubated chick embryos

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Junctional complexes have been investigated in the epiblast of young chick embryos by examination of freeze-fracture replicas and of sections of comparable specimens stained with lanthanum nitrate. By means of freeze-fracture, tight junctions were shown to be present in the unincubated embryo (stage 1 of Hamburger and Hamilton). The number of ridges or grooves was found to vary between 2 and 10 near the dorsal border, whereas isolated ridges were found more ventrally. Lanthanum was unable to penetrate between the cells in the region of the dorsally situated tight junctions. Similar tight junctions were found in incubated embryos (stage 3) examined by both techniques. Tight junctions were also seen in cleavage (pre-laying) embryos examined in section.

Gap junctions were extremely uncommon in unincubated embryos, though occasional aggregates of gap junction particles were seen on the lateral cell membranes close to the dorsal surface. In only one instance were associated pits visible. By contrast, gap junctions were more frequently encountered by stage 3, and these junctions possessed both pits and particles. Desmosomes were never seen in the freeze-fracture replicas at either stages 1 or 3, though structures which might be developing desmosomes were visible in sections.

The functions of both the tight and gap junctions in the young chick embryo are discussed. The results are also considered in relation to recent theories about the way in which gap junctions are formed.

We are most grateful to Mr. R. Moss and Miss D. Bailey for technical assistance, and to Mrs. J. Astafiev for drawing Fig. 1. We would also like to thank Dr. R. A. Weiss and Dr. Susan Pegrum of the Imperial Cancer Research Fund Laboratories for helpful discussions. — We also acknowledge the generous support of the Wellcome Trust, The Medical Research Council and the Joint Standing Research Committee of St. Mary's Hospital in developing freezefracture facilities.