Gal4-Gene-Dependent Alterations of Embryo Development and Cell Growth in Primary Culture of Sea Urchins

• Published in: Marine biotechnology (New York, N.Y.) Vol. 4; no. 5; pp. 480 - 486
• Main Authors: Bulgakov, V. P, Odintsova, N. A, Plotnikov, S. V, Kiselev, K. V, Zacharov, E. V, Zhuravlev, Y. N
• Format: Journal Article
• Language: English
• Published: United States Springer-Verlag 01-10-2002; Springer Nature B.V
• Subjects: Animal embryos; Biotechnology; Blastula; Cell culture; Cell growth; Cell number; Cells; Cultivation; Cultures; Deoxyribonucleic acid; DNA; DNA biosynthesis; Echinoidea; embryogenesis; Embryos; GAL4 gene; Gene expression; genes; Marine; Marine invertebrates; Nucleotide sequence; PCR; Plasmids; Polymerase chain reaction; reverse transcriptase polymerase chain reaction; RNA-directed DNA polymerase; Sea urchins; Structures; Synthesis; Teratoma; Transcription; Yeast; Yeasts
• ISSN: 1436-2228; 1436-2236
• DOI: 10.1007/s10126-002-0036-7

Primary cell cultures from sea urchins have a low proliferative level that prevents the establishment of long-term cultures. To increase expression levels of the genes regulating cell growth in sea urchins, and thus enhance cell growth, we used the transcriptional activator gene Gal4 found earlier in yeast. Sea urchin embryos were treated with plasmid DNA containing the Gal4 gene. Expression of the transgene was confirmed by reverse transcriptase polymerase chain reaction. When the fully functional gene was used, embryos effectively formed teratoma-like structures after 50 to 55 hours of cultivation. In contrast, the Gal4 gene, devoid of acidic activating regions, possessed little activity as a teratogen. The Gal4-treated cells in blastula-derived culture showed higher DNA synthesis and higher proliferative activity than control cells. We suggest that formation of the teratoma-like structures in embryos, activation of DNA synthesis, and significant increase of cell number in embryo-derived cell cultures could be attributed to Gal4 gene action.