[Influence of the activator of transcription GAL4 on growth and development of embryos and embryonic cells in primary cultures of sand dollar].

Institute of Marine Biology, Far Eastern Branch of the Russian Academy of Sciences, ul. Pal'chevskoro 17, Vladivostok, 690041 Russia.
Ontogenez 01/2003; 34(4):267-72. DOI: 10.1023/A:1024992415489
Source: PubMed

ABSTRACT In order to solve many tasks of biotechnology, constant lines of the cells of marine invertebrates with a high growth potential are required, which are absent at present. We used the universal activator of transcription gal4 to change the degree of expression of genes of growth factors in embryonic sea urchin cells and, thereby, increase their proliferative activity. The fertilized sea urchin eggs and dissociated embryonic cells at the blastula stage were treated with plasmids containing both the functional gene gal4 and the gene devoid of the regions encoding the activator domain. The transfection of embryonic sea urchin eggs with the functional gene led to cell dedifferentiation and formation of tumor-like structures in the embryos or increased number of embryonic cells in culture. In the cells obtained from the transfected embryos, the pigments were found within two months of cultivation, whose absorption spectrum coincided with that of echinochrome.

  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The quinone pigments of sea urchins, specifically echinochrome and spinochromes, are known for their effective antioxidant, antibacterial, antifungal, and antitumor activities. We developed in vitro technology for inducing pigment differentiation in cell culture. The intensification of the pigment differentiation was accompanied by a simultaneous decrease in cell proliferation. The number of pigment cells was two-fold higher in the cells cultivated in the coelomic fluids of injured sea urchins than in those intact. The possible roles of the specific components of the coelomic fluids in the pigment differentiation process and the quantitative measurement of the production of naphthoquinone pigments during cultivation were examined by MALDI and electrospray ionization mass spectrometry. Echinochrome A and spinochrome E were produced by the cultivated cells of the sand dollar Scaphechinus mirabilis in all tested media, while only spinochromes were found in the cultivated cells of another sea urchin, Strongylocentrotus intermedius. The expression of genes associated with the induction of pigment differentiation was increased in cells cultivated in the presence of shikimic acid, a precursor of naphthoquinone pigments. Our results should contribute to the development of new techniques in marine biotechnology, including the generation of cell cultures producing complex bioactive compounds with therapeutic potential.
    Marine Drugs 07/2014; 12(7):3874-3891. DOI:10.3390/md12073874 · 3.51 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Many problems of modern virology, biotechnology and ecology cannot be approached without the use of permanent cell lines from marine invertebrates. Tissues with high growth potential are useful for establishment of such cell lines. We estimated the prevalence of naturally occurring tumor-like lesions and neoplasms in bivalve molluscs from the Sea of Japan with the focus on cell mitotic activity and tried to induce similar formations in sea urchins in the laboratory. Inspecting nearly 3000 bivalves of eight species, we have detected the tumor-like lesions occurring in 0–15% of the total number of molluscs that were studied. Many of the molluscan tumor-like lesions had a low mitotic index (MI, 0.01–0.3%). Neoplasms with the MI 3–12% were met only in Mytilus and Pecten. Besides a search for naturally occurring tumor-like lesions and neoplasms in molluscs, we used modulation of gene expression by genetically engineered constructs bearing foreign genes, the yeast transcriptional activator gal4 gene or the agrobacterial rol oncogenes, in sea urchin embryos and sea urchin embryonic cells. We failed to develop a potential permanent cell line; however, the results obtained allow us to assert that some tumor-like formations can be useful objects for establishment of permanent cell lines. Only cells of malignant mussel hemocytes and developmental anomalies in sea urchin embryos transformed by the yeast gal4 gene were involved in active proliferation. Our findings suggest that the transformed by the yeast gal4 gene embryonic cells of sea urchins may be as a suitable model for future studies.
    Journal of Experimental Marine Biology and Ecology 10/2011; 407(2):241-249. DOI:10.1016/j.jembe.2011.06.020 · 2.48 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The review surveys own and the literature data on the plasticity of marine invertebrate stem cells. Stem and embryonic cell cultures of marine invertebrates are a novel model system characterized by a high level of physiological and synthetic processes. The production of biologically active substances in vitro may be an alternative to chemical synthesis and aquaculture. The factors involved in determination and maintenance of the pluripotency of marine invertebrate stem cells have been analyzed. The technology of the directed differentiation of marine invertebrate stem cells into certain functionally active cells in vitro embraces the use of different growth factors, various natural and artificial substrates, and unique bioactive compounds from marine invertebrate tissues. To increase the expression levels of regulatory genes, we applied genetically engineered constructions with foreign genes. The regulation of growth and differentiation of marine invertebrate stem cells opens new prospective uses for their application in marine biotechnology and is helpful for research in developmental biology.
    Cell and Tissue Biology 10/2009; 3(5):403-408. DOI:10.1134/S1990519X09050010

Full-text (3 Sources)

Available from
Jun 3, 2014