title: Using Morpholinos to Examine Gene Function During Fin Regeneration.
abstract: In the zebrafish regenerating fin, specific gene-targeting morpholinos have been widely utilized to assess gene function. Unlike in embryos, injection of standard morpholinos in the adult regenerating fin is not sufficient for cellular uptake. Rather, morpholinos are first injected extracellularly into the blastemal compartment, followed by electroporation for cellular uptake. Knockdown phenotypes are evaluated 1-4 days post electroporation. This chapter provides a description of the reagents, equipment, and procedure for successful injection and electroporation of morpholinos into the regenerating fin.
authors.affiliation: Departments of Anatomy and Cell Biology and Opthamology, Wayne State University School of Medicine, Wayne State University, Detroit, MI, USA.
authors.forename: Ryan
authors.initials: R
authors.lastname: Thummel
authors.affiliation: Department of Biological Sciences, Lehigh University, Iacocca Hall, 111 Research Drive, D222, Bethlehem, PA, 18015, USA. mki3@lehigh.edu.
authors.forename: M
authors.initials: M
authors.lastname: Kathryn Iovine
mesh_terms: D058500:Animal Fins / Q000254:growth & development*; D000818:Animals; D018274:Electroporation / Q000379:methods; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D008845:Microinjections; D060172:Morpholinos / Q000008:administration & dosage / Q000235:genetics*; D010641:Phenotype; D012038:Regeneration / Q000235:genetics*; D015027:Zebrafish / Q000235:genetics; D029961:Zebrafish Proteins / Q000235:genetics
chemical_list: D060172:Morpholinos; D029961:Zebrafish Proteins
keywords: Electroporation; Fin regeneration; Gene knockdown; Zebrafish