Tenascin-C expression in the trunk of wild-type, cyclops and floating head zebrafish embryos

Abstract

The function and the regulation of the expression of the extracellular matrix molecule tenascin-C during embryonic development are still unclear. In the present study, the expression of tenascin-C was analyzed in the trunk of zebrafish at the end of the first embryonic day. An antiserum raised against a zebrafish tenascin-C (TN-C) fusion protein reacted with 220 (doublet), 200, and 160 KD peptides. In situ hybridization showed that in the zebrafish wild-type embryo, tn-c mRNA was expressed by somites, neural crest cells, roof plate, notochord, hypochord, and tail fin bud. Thus, the expression of tn-c mRNA is an excellent marker for the differentiation of most zebrafish trunk structures. Immunolabelling with the anti-TN-C antibody was detected in the migratory pathway of neural crest cells and in the intersomitic furrows. In situ hybridization analysis of the zebrafish cyclops mutants, lacking the midline floor plate cells, showed normal expression of tn-c mRNA in all trunk structures. Analysis of the floating-head mutant, lacking the notochord, showed that tn-c mRNA expression in neural crest cells, roof plate, and tail fin bud is normal, but it is defective in the somites. By showing that the noctochord, but not the floor plate, cells are required for normal tn-c expression in the trunk, this work provides new information on the role played by the embryonic axial structures in the regulation of the expression of tn-c during the development of zebrafish and allows new conclusions about somite patterning in the cyclops and floating-head zebrafish mutants.