4.4 Article

A Method for Microinjection of Patiria minata Zygotes

Journal

JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
Volume -, Issue 91, Pages -

Publisher

JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/51913

Keywords

Developmental Biology; Issue 91; Embryology; Patiria miniata; sea star; echinoderm; development; gene regulatory networks; microinjection; gene expression perturbation; antisense oligonucleotide; reporter expression

Funding

  1. National Science Foundation [IOS 0844948, IOS 1024811]

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Echinoderms have long been a favorite model system for studies of reproduction and development, and more recently for the study of gene regulation and evolution of developmental processes. The sea star, Patiria miniata, is gaining prevalence as a model system for these types of studies which were previously performed almost exclusively in the sea urchins, Strongylocentrotus purpuratus and Lytechinus variegatus. An advantage of these model systems is the ease of producing modified embryos in which a particular gene is up or downregulated, labeling a group of cells, or introducing a reporter gene. A single microinjection method is capable of creating a wide variety of such modified embryos. Here, we present a method for obtaining gametes from P. miniata, producing zygotes, and introducing perturbing reagents via microinjection. Healthy morphant embryos are subsequently isolated for quantitative and qualitative studies of gene function. The availability of genome and transcriptome data for this organism has increased the types of studies that are performed and the ease of executing them.

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