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Genes 2018, 9(4), 197; https://doi.org/10.3390/genes9040197

Transgenic Xenopus laevis Line for In Vivo Labeling of Nephrons within the Kidney

1
Department of Pediatrics, Pediatric Research Center, UTHealth McGovern Medical School, Houston, TX 77030, USA
2
Program in Biochemistry and Cell Biology, Rice University, Houston, TX 77005, USA
3
Program in Genes and Development, MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences, Houston, TX 77030, USA
4
National Xenopus Resource and Eugene Bell Center for Regenerative Biology and Tissue Engineering, Marine Biological Laboratory, Woods Hole, MA 02543, USA
5
Department of Genetics, University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA
6
Department of Molecular Medicine and Pathology, University of Auckland, Auckland 1010, New Zealand
7
Program in Genetics and Epigenetics, MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences, Houston, TX 77030, USA
8
Program in Biochemistry and Cell Biology, MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences, Houston, TX 77030, USA
These authors contributed equally to this work.
Current address: Centre for Craniofacial and Regenerative Biology, Dental Institute, King’s College London, London WC2R 2LS, UK.
§
Current address: Cancer and Developmental Biology Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD 21702, USA.
*
Author to whom correspondence should be addressed.
Received: 28 February 2018 / Revised: 29 March 2018 / Accepted: 4 April 2018 / Published: 6 April 2018
(This article belongs to the Special Issue DNA Variations in Evolution and Human Diseases)
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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. View Full-Text
Keywords: cdh17; pronephros; mesonephros; kidney; nephron; live imaging; primary culture; Xenopus; transgenic cdh17; pronephros; mesonephros; kidney; nephron; live imaging; primary culture; Xenopus; transgenic
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Corkins, M.E.; Hanania, H.L.; Krneta-Stankic, V.; DeLay, B.D.; Pearl, E.J.; Lee, M.; Ji, H.; Davidson, A.J.; Horb, M.E.; Miller, R.K. Transgenic Xenopus laevis Line for In Vivo Labeling of Nephrons within the Kidney. Genes 2018, 9, 197.

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