Development of Sea Urchins, Ascidians, and Other Invertebrate Deuterostomes: Experimental Approaches

Overview: Echinoderms and other invertebrate deuterostomes as experimental material for the study of cell and developmental biology; I. Procurement, Maintenance and Culture of Oocytes, Embryos, Larvae, and Adults; Care and maintenance of adult echinoderms (species, breeding seasons, suppliers, care and handling, aquarium maintenance,large-scale aquaculture); Echinoderm eggs and embryos: Procurement and culture (small and large scale); Culture of echinoderm larvae through metamorphosis; Isolation and maturation of echinoderm oocytes; Procurement and culture of ascidian embryos; Culture of adult ascidians and ascidian genetics; Hemichordate embryos - procurement, culture, and basic methods; Cephalochordate embryos - procurement, culture, and basic methods. II. Embryological
Approaches; Microinjection into eggs, blastomeres, and the blastocoel; Blastomere isolation and transplantation; Isolation and culture of micromeres and PMCs; Transplantation and ablation of cells at later stages; Cell adhesion assays. III. Cell Biological Approaches; Immunostaining; Light microscopy of embryos; Electron microscopy of embryos; Calcium imaging; Imaging components of cells with fluorescent probes; Isolating extracellular matrices; Isolating organelles; Measuring ion fluxes in gametes. IV. Molecular Biological Approaches; Isolating DNA, RNA, polysomes, and protein; m-RNA detection by in situ hybridization and RT-PCR; Using reporters to study cis-regulatory elements; Nuclear extracts, footprinting mobility shift assays, affinity purification; Expression of exogenous mRNAs; Disruption of gene function using antisense morpholinos and function-blocking antibodies; Generation and use of transgenic ascidian embryos. V. Genomics; Genomic resources for echinoderms: Sequence databases, arrayed libraries and filters; Genomic resources for ascidians: Sequence/expression databases and genome projects; Gene network analysis. VI. Echinoderm Eggs and Embryos in the Teaching Lab; Simple and reliable laboratory exercises. VII. Appendices.