title: Technique to Target Microinjection to the Developing Xenopus Kidney.
abstract: The embryonic kidney of Xenopus laevis (frog), the pronephros, consists of a single nephron, and can be used as a model for kidney disease. Xenopus embryos are large, develop externally, and can be easily manipulated by microinjection or surgical procedures. In addition, fate maps have been established for early Xenopus embryos. Targeted microinjection into the individual blastomere that will eventually give rise to an organ or tissue of interest can be used to selectively overexpress or knock down gene expression within this restricted region, decreasing secondary effects in the rest of the developing embryo. In this protocol, we describe how to utilize established Xenopus fate maps to target the developing Xenopus kidney (the pronephros), through microinjection into specific blastomere of 4- and 8-cell embryos. Injection of lineage tracers allows verification of the specific targeting of the injection. After embryos have developed to stage 38 - 40, whole-mount immunostaining is used to visualize pronephric development, and the contribution by targeted cells to the pronephros can be assessed. The same technique can be adapted to target other tissue types in addition to the pronephros.
authors.affiliation: Department of Pediatrics, Pediatric Research Center, University of Texas McGovern Medical School.
authors.forename: Bridget D
authors.initials: BD
authors.lastname: DeLay
authors.affiliation: Department of Pediatrics, Pediatric Research Center, University of Texas McGovern Medical School; Program in Genes & Development, University of Texas Graduate School of Biomedical Sciences.
authors.forename: Vanja
authors.initials: V
authors.lastname: Krneta-Stankic
authors.affiliation: Department of Pediatrics, Pediatric Research Center, University of Texas McGovern Medical School; Program in Genes & Development, University of Texas Graduate School of Biomedical Sciences; Program in Cell & Regulatory Biology, University of Texas Graduate School of Biomedical Sciences; Department of Genetics, University of Texas MD Anderson Cancer Center; Rachel.K.Miller@uth.tmc.edu.
authors.forename: Rachel K
authors.initials: RK
authors.lastname: Miller
mesh_terms: D000818:Animals; D018507:Gene Expression Regulation, Developmental; D008845:Microinjections / Q000379:methods*; D060910:Pronephros; D014981:Xenopus; D014982:Xenopus laevis / Q000033:anatomy & histology / Q000254:growth & development