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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.
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Bauer,
The cleavage stage origin of Spemann's Organizer: analysis of the movements of blastomere clones before and during gastrulation in Xenopus.
1994, Pubmed,
Xenbase
Bauer,
The cleavage stage origin of Spemann's Organizer: analysis of the movements of blastomere clones before and during gastrulation in Xenopus.
1994,
Pubmed
,
Xenbase
Brändli,
Towards a molecular anatomy of the Xenopus pronephric kidney.
1999,
Pubmed
,
Xenbase
Carroll,
Synergism between Pax-8 and lim-1 in embryonic kidney development.
1999,
Pubmed
,
Xenbase
Colman,
The stability and movement of mRNA in Xenopus oocytes and embryos.
1986,
Pubmed
,
Xenbase
Davidson,
Integrin alpha5beta1 and fibronectin regulate polarized cell protrusions required for Xenopus convergence and extension.
2006,
Pubmed
,
Xenbase
Etkin,
Replication of injected DNA templates in Xenopus embryos.
1987,
Pubmed
,
Xenbase
Huang,
Blastomeres show differential fate changes in 8-cell Xenopus laevis embryos that are rotated 90 degrees before first cleavage.
1998,
Pubmed
,
Xenbase
Karpinka,
Xenbase, the Xenopus model organism database; new virtualized system, data types and genomes.
2015,
Pubmed
,
Xenbase
Krneta-Stankic,
Xenopus: leaping forward in kidney organogenesis.
2016,
Pubmed
,
Xenbase
Lienkamp,
Vertebrate kidney tubules elongate using a planar cell polarity-dependent, rosette-based mechanism of convergent extension.
2012,
Pubmed
,
Xenbase
Lyons,
Requirement of Wnt/beta-catenin signaling in pronephric kidney development.
2009,
Pubmed
,
Xenbase
Miller,
Pronephric tubulogenesis requires Daam1-mediated planar cell polarity signaling.
2011,
Pubmed
,
Xenbase
Moody,
Fates of the blastomeres of the 16-cell stage Xenopus embryo.
1987,
Pubmed
,
Xenbase
Moody,
Segregation of fate during cleavage of frog (Xenopus laevis) blastomeres.
1990,
Pubmed
,
Xenbase
Moody,
Fates of the blastomeres of the 32-cell-stage Xenopus embryo.
1987,
Pubmed
,
Xenbase
Raciti,
Organization of the pronephric kidney revealed by large-scale gene expression mapping.
2008,
Pubmed
,
Xenbase
Taira,
Expression of the LIM class homeobox gene Xlim-1 in pronephros and CNS cell lineages of Xenopus embryos is affected by retinoic acid and exogastrulation.
1994,
Pubmed
,
Xenbase
Ubbels,
Evidence for a functional role of the cytoskeleton in determination of the dorsoventral axis in Xenopus laevis eggs.
1983,
Pubmed
,
Xenbase
Uochi,
The Na+,K+-ATPase alpha subunit requires gastrulation in the Xenopus embryo.
1997,
Pubmed
,
Xenbase
Vize,
Development of the Xenopus pronephric system.
1995,
Pubmed
,
Xenbase
Vize,
The chloride conductance channel ClC-K is a specific marker for the Xenopus pronephric distal tubule and duct.
2003,
Pubmed
,
Xenbase
Vize,
Model systems for the study of kidney development: use of the pronephros in the analysis of organ induction and patterning.
1997,
Pubmed
,
Xenbase
Wallingford,
Precocious expression of the Wilms' tumor gene xWT1 inhibits embryonic kidney development in Xenopus laevis.
1998,
Pubmed
,
Xenbase
Weber,
Regulation and function of the tissue-specific transcription factor HNF1 alpha (LFB1) during Xenopus development.
1996,
Pubmed
,
Xenbase
Zhou,
Proximo-distal specialization of epithelial transport processes within the Xenopus pronephric kidney tubules.
2004,
Pubmed
,
Xenbase
