Spatially deranged though temporally correct expression of Strongylocentrotus purpuratus actin gene fusion in transgenic embryos of a different sea urchin family.
We report the unexpected observation that cis-regulatory sequences of a Strongylocentrotus purpuratus actin gene, which direct a particular, lineage-specific pattern of embryonic expression, confer a completely different spatial pattern of expression when introduced into embryos of another sea urchin species. We utilized a fusion gene construct in which the bacterial chloramphenicol acetyl transferase (CAT) reporter gene is driven by CyIIIa actin regulatory sequences. We previously showed that the regulatory region that is included suffices to promote the accumulation of CAT mRNA in transgenic S. purpuratus embryos, on the same developmental schedule and in the same embryonic region, the aboral ectoderm, in which the CyIIIa actin gene is normally expressed (Flytzanis et al. 1987; Hough-Evans et al. 1987). When injected into zygotes of Lytechinus variegatus, which belongs to a different echinoid family, the expected temporal pattern of expression of CAT enzyme was observed. Thus, in both S. purpuratus and L. variegatus embryos, expression is activated at the early blastula stage, although this stage is attained several hours sooner in L. variegatus embryo cultures. Similar kinetics of CAT enzyme accumulation were obtained whether the gene was introduced directly into the L. variegatus zygote nucleus or into the cytoplasm. However, when examined by in situ hybridization, the transgenic L. variegatus embryos were found to display a totally new pattern of CAT mRNA accumulation. Copious CAT transcripts were detected not only in aboral ectoderm cells, but also in skeletogenic mesenchyme cells, gut cells, and oral ectoderm, all cell types that in the transgenic S. purpuratus controls are invariably devoid of detectable CAT transcripts.
Franks
R R
RR
Division of Biology, California Institute of Technology, Pasadena 91125.
Hough-Evans
B R
BR
Britten
R J
RJ
Davidson
E H
EH
D000123:Acetyltransferases / Q000235:genetics; D000199:Actins / Q000235:genetics*; D000818:Animals; D030801:Animals, Genetically Modified / Q000196:embryology* / Q000235:genetics; D002467:Cell Nucleus / Q000502:physiology; D015500:Chloramphenicol O-Acetyltransferase; D003593:Cytoplasm / Q000502:physiology; D004475:Ectoderm / Q000502:physiology; D005786:Gene Expression Regulation; D005809:Genes, Regulator; D008845:Microinjections; D009693:Nucleic Acid Hybridization; D012045:Regulatory Sequences, Nucleic Acid; D012617:Sea Urchins / Q000196:embryology* / Q000235:genetics*; D014018:Tissue Distribution
D000199:Actins; D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase