Molecular characterization of a heat shock cognate cDNA of zebrafish, hsc70, and developmental expression of the...

To elucidate the potential role of the hsp70 gene family in developmental processes in vertebrates, we chose to study the expression of one of these genes in zebrafish. A zebrafish gastrula cDNA library was screened with a Pleurodeles waltl hsp70 cDNA probe. A 2.3-kb cDNA was thus isolated and sequenced. The predicted amino acid sequence contained an open reading frame encoding for a 649-amino acid polypeptide. Sequence analysis showed strong homology with hsp70-related gene sequences in other species; in particular, the strongest homology was found with the cognate members of this family. Tests of heat inducibility revealed that transcripts were expressed at normal temperature, but the level of transcript expression increased after heat shock. Moreover, experiments of the neosynthesis of total proteins in heat shock conditions and corresponding immunoblotting assays showed that 24-h-stage embryos are able to respond to heat shock. The quantity of 70 kDa proteins, recognized by a specific antibody of the HSP/C70 protein family, is expressed in control condition and increased significantly after heat shock. Furthermore, Northern blot analysis of transcript expression showed that the corresponding mRNAs were detected throughout embryonic development in the absence of any heat shock. Our clone, named hsc70, thus corresponded to a cognate member of the hsp70 gene family, expressed under normal conditions during development, but also heat inducible. The spatio-temporal pattern of transcripts during development was determined by in situ hybridization on wholemount embryos at different stages. As a maternal RNA, hsc70 mRNA was uniformly present in the embryo, up to the end of gastrulation. Later, a tissue-specific enrichment of hsc70 transcripts was detected in the central nervous system (CNS) and in a fraction of the somites. These results suggest that the hsc70 gene may be involved in developmental differentiation events.