Affiliation:
1. Division of Biology 156-29, California Institute of Technology, Pasadena,CA 91125, USA
Abstract
New secondary mesenchyme specific genes, expressed exclusively in pigment cells, were isolated from sea urchin embryos using a differential screening of a macroarray cDNA library. The comparison was performed between mRNA populations of embryos having an expansion of the endo-mesodermal territory and embryos blocked in secondary mesenchyme specification. To be able to isolate transcripts with a prevalence down to five copies per cell, a subtractive hybridization procedure was employed. About 400 putative positive clones were identified and sequenced from the 5′ end. Gene expression analysis was carried out on a subset of 66 clones with real time quantitative PCR and 40 clones were positive. This group of clones contained sequences highly similar to: the transcription factor glial cells missing(gcm); the polyketide synthase gene cluster (pks-gc); three different members of the flavin-containing monooxygenase gene family(fmo); and a sulfotransferase gene (sult). Using whole mount in situ hybridization, it was shown that these genes are specifically expressed in pigment cells. A functional analysis of the S. purpuratus pks and of one S. purpuratus fmo was carried out using antisense technology and it was shown that their expression is necessary for the biosynthesis of the sea urchin pigment echinochrome. The results suggest that S. purpuratus pks, fmo and sult could belong to a differentiation gene battery of pigment cells.
Publisher
The Company of Biologists
Subject
Developmental Biology,Molecular Biology
Reference91 articles.
1. Akiyama, Y., Hosoya, T., Poole, A. M. and Hotta, Y.(1996). The gcm-motif: a novel DNA-binding motif conserved in Drosophila and mammals. Proc. Natl. Acad. Sci. USA93,14912-14916.
2. Alfonso, T. B. and Jones, B. W. (2002). gcm2 promotes glial cell differentiation and is required with glial cells missing for macrophage development in Drosophila. Dev. Biol. 248,369-383.
3. Altschul, S. F., Madden, T. L., Schaffer, A. A., Zhang, J.,Zhang, Z., Miller, W. and Lipman, D. J. (1997). Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 25,3389-3402.
4. Angerer, L. M. and Angerer, R. C. (2000). Animal-vegetal axis patterning mechanisms in the early sea urchin embryo. Dev. Biol. 218,1-12.
5. Arenas-Mena, C., Cameron, A. R. and Davidson, E. H.(2000). Spatial expression of Hox cluster genes in the ontogeny of a sea urchin. Development127,4631-4643.
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