Graduation Year

2012

Document Type

Dissertation

Degree

Ph.D.

Degree Granting Department

Biology (Cell Biology, Microbiology, Molecular Biology)

Major Professor

James R. Garey

Keywords

Brittle Star, Gastrula, Gene Regulatory Networks, Sea Urchin, Transcription Factors

Abstract

The gastrula stage represents the point in development at which the three primary germ layers diverge. At this point the gene regulatory networks that specify the germ layers are established and the genes that define the differentiated states of the tissues have begun to be activated. These networks have been well characterized in sea urchins, but not in other echinoderms. Embryos of the brittle star Ophiocoma wendtii share a number of developmental features with sea urchin embryos, including the ingression of mesenchyme cells that give rise to an embryonic skeleton. Notable differences are that no micromeres are formed during cleavage divisions and no pigment cells are formed during development to the pluteus larva stage. More subtle changes in timing of developmental events also occur. To explore the molecular basis for the similarities and differences between these two echinoderms, the gastrula transcriptome of Ophiocoma wendtii was sequenced and characterized.

I identified brittle star transcripts that correspond to 3385 genes in existing databases, including 1863 genes shared with the sea urchin Strongylocentrotus purpuratus gastrula transcriptome. I have characterized the functional classes of genes present in the transcriptome and compared them to those found in sea urchin. I then examined which members of the germ-layer specific gene regulatory networks (GRNs) of S. purpuratus are expressed in the O. wendtii gastrula. The results indicate that there is a shared "genetic toolkit" central to the echinoderm gastrula, a key stage in embryonic development, though there are also differences that reflect changes in developmental processes.

The brittle star expresses genes representing all functional classes at the gastrula stage. Brittle stars and sea urchins have comparable numbers of each class of genes, and share many of the genes expressed at gastrula. Examination of the brittle star genes whose sea urchin orthologs are utilized in germ layer specification reveals a relatively higher level of conservation of key regulatory components compared to the overall transcriptome. I also identify genes that were either lost or whose temporal expression has diverged from that of sea urchins. Overall, the data suggest that embryonic skeleton formation in sea urchins and brittle stars represents convergent evolution by independent cooptation of a shared pathway utilized in adult skeleton formation.

Transcription factors are of central importance to both development and evolution. Patterns of their expression and interactions form the gene regulatory networks which control the building of the embryonic body. Alterations in these patterns can result in the construction of altered bodies. To help increase understanding of this process, I compared the transcription factor mRNAs present in early gastrula-stage embryos of the brittle star Ophiocoma wendtii to those found in two species of sea urchins and a starfish. Brittle star homologs were found for one third of the transcription factors in the sea urchin genome and half of those that are expressed at equivalent developmental stages in sea urchins and starfish. Overall, the patterns of transcription factors found and not found in brittle star resemble those of other echinoderms, with the differences largely consistent with morphological differences. This study provides further evidence for the existence of deeply conserved developmental genetic processes, with various elements shared among echinoderms, deuterostomes, and metazoans.

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