Dev Growth Differ 1990 Feb 01;321:33-39. doi: 10.1111/j.1440-169X.1990.00033.x.

A New Technique for Introducing Anti-Fibronectin Antibodies and Fibronectin-Related Synthetic Peptides into the Blastulae of the Sea Urchin, Clypeaster japonicus.: (fibronectin/synthetic peptide/cell migration/sea urchin/primary mesenchyme cell).

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Migration of primary mesenchyme cells (PMCs) of the sea urchin, Clypeaster japonicus, was examined in vivo by introducing anti-fibronectin (FN) IgG, and FN-related synthetic peptides, Gly-Arg-Gly-Asp-Ser-Pro-Cys, Gly-Arg-Gly-Asp-Ser, and Arg-Gly-Asp-Ser (RGDS) into the blastocoel by a new technique. In this technique the embryos are treated with cytochalasin B (CB) and part of the presumptive PMCs (PPMCs) is removed, leaving a hole in the vegetal plate. Then macromolecules are introduced into the blastocoel through this hole. Their introduction was confirmed by introducing them with polystyrene beads as a marker. The hole closes soon after introduction of the materials, and so these materials remain in the blastocoel. After this treatment, blastulae had fewer PMCs, because of partial loss of PPMCs, but their morphogenesis proceeded normally. Introduction of anti-FN IgG or the synthetic peptides inhibited PMC migration in vivo, and this inhibition was associated with failure of the PMCs to form cell processes. These results indicate that sea urchin PMCs use the RGDS amino acid sequence in the FN molecule for migration in vivo.

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