Transgenic Xenopus laevis line for in vivo labeling of nephrons within the kidney
Corkins, Mark E.
Hanania, Hannah L.
DeLay, Bridget D.
Pearl, Esther J.
Davidson, Alan J.
Horb, Marko E.
Miller, Rachel K.
MetadataShow full item record
Keywordcdh17; Pronephros; Mesonephros; Kidney; Nephron; Live imaging; Primary culture; Xenopus; Transgenic
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.
© The Author(s), 2018. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Genes 9 (2018): 197, doi:10.3390/genes9040197.
Suggested CitationGenes 9 (2018): 197
The following license files are associated with this item:
Showing items related by title, author, creator and subject.
Development of xenopus resource centers : the national xenopus resource and the european xenopus resource center Pearl, Esther J.; Grainger, Robert M.; Guille, Matthew; Horb, Marko E. (2011-07)Xenopus is an essential vertebrate model system for biomedical research that has contributed to important discoveries in many disciplines, including cell biology, molecular biology, physiology, developmental biology and ...
Nguyen, Phuong A.; Field, Christine M.; Mitchison, Timothy J. (American Society for Cell Biology, 2018-03-23)The cleavage furrow in Xenopus zygotes is positioned by two large microtubule asters that grow out from the poles of the first mitotic spindle. Where these asters meet at the midplane, they assemble a disk-shaped interaction ...
Ratzan, Wil; Falco, Rosalia; Salanga, Cristy; Salanga, Matthew C.; Horb, Marko E. (Elsevier, 2016-03-15)Completion of the Xenopus laevis genome sequence from inbred J strain animals has facilitated the generation of germline mutant X. laevis using targeted genome editing. In the last few years, numerous reports have demonstrated ...