Publication Date:
2022-05-25
Description:
© 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.
Description:
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.
Description:
These studies were supported by a postdoctoral fellowship from the National Kidney Foundation (FLB1628 to R.K.M), a National Institutes of Health (NIH) KO1 grant (K01DK092320 to R.K.M.), and startup funding from UTHealth McGovern Medical School’s Department of Pediatrics (to R.K.M.).
Keywords:
cdh17
;
Pronephros
;
Mesonephros
;
Kidney
;
Nephron
;
Live imaging
;
Primary culture
;
Xenopus
;
Transgenic
Repository Name:
Woods Hole Open Access Server
Type:
Article
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