Frog lim-1-like protein is expressed predominantly in the nervous tissue, gonads, and early embryos of the bivalve mollusc Mytilus galloprovincialis.

Abstract

In a few well-known cases, the biological consequences of the disruption of lim-1 homeodomain (HD) genes have demonstrated the important roles of these genes in vertebrate development, especially in the nervous tissue, kidney, and gonads. Functional assay approaches require information not only about lim-1 gene organization, but also about properties and tissue localization of Lim-1 proteins. Although lim-1 genes have been identified in certain phyla of invertebrates, no information is available on Lim-1 proteins and genes in bivalve molluscs. Our study represents the beginning stage of identification of the Lim-1-related proteins in marine bivalves. Using antibodies against the C-terminal region of the Xenopus laevis Lim-1 protein, we describe cross-reactive antigen patterns in adults and early embryos of the mussel Mytilus galloprovincialis, as well as in sea urchin and chick embryos. In adult mussels, nervous ganglia and gonads display the most prominent Lim-1 immunoreactivity. Further, the antibodies verified the prediction that mussel Lim-1 antigens, like Lim-1 HD proteins in general, can be localized in the nucleus. Moreover, antibody detection allowed us to identify the Lim-1-like antigens in unfertilized mature eggs, as well as in very early embryos of bivalve molluscs and sea urchins (Strongylocentrotus purpuratus). In mussel eggs and embryos, Lim-1 antigens are expressed in multiple forms (40, 45, and 65 kDa), as detected by SDS-PAGE followed by Western blot. Taken together, the observations emphasize the conservation of the Lim-1 protein expression pattern in the nervous tissue and gonads of different animal groups, and demonstrate that Lim-1-like polypeptides can be maternally accumulated in eggs and, therefore, are present in very early embryos before zygotic expression of the genes begins.