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XB-ART-54809
Genes (Basel) April 6, 2018; 9 (4):
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Transgenic Xenopus laevis Line for In Vivo Labeling of Nephrons within the Kidney.

Corkins ME , Hanania HL , Krneta-Stankic V , DeLay BD , Pearl EJ , Lee M , Ji H , Davidson AJ , Horb ME , Miller RK .


Abstract
Xenopus laevis embryos are an established model for studying kidney development. The nephron structure and genetic pathways that regulate nephrogenesis are conserved between Xenopus and humans, allowing for the study of human disease-causing genes. Xenopus embryos are also amenable to large-scale screening, but studies of kidney disease-related genes have been impeded because assessment of kidney development has largely been limited to examining fixed embryos. To overcome this problem, we have generated a transgenic line that labels the kidney. We characterize this cdh17:eGFP line, showing green fluorescent protein (GFP) expression in the pronephric and mesonephric kidneys and colocalization with known kidney markers. We also demonstrate the feasibility of live imaging of embryonic kidney development and the use of cdh17:eGFP as a kidney marker for secretion assays. Additionally, we develop a new methodology to isolate and identify kidney cells for primary culture. We also use morpholino knockdown of essential kidney development genes to establish that GFP expression enables observation of phenotypes, previously only described in fixed embryos. Taken together, this transgenic line will enable primary kidney cell culture and live imaging of pronephric and mesonephric kidney development. It will also provide a simple means for high-throughput screening of putative human kidney disease-causing genes.

PubMed ID: 29642376
PMC ID: PMC5924539
Article link: Genes (Basel)
Grant support: [+]

Species referenced: Xenopus laevis
Genes referenced: cdh17 daam1


Article Images: [+] show captions
References [+] :
BARTH, Differentiation of cells of the Rana pipiens gastrula in unconditioned medium. 1998, Pubmed