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

• Published in: Genes 2018-04, Vol.9 (4), p.197
• Main Authors: Corkins, Mark E, Hanania, Hannah L, Krneta-Stankic, Vanja, DeLay, Bridget D, Pearl, Esther J, Lee, Moonsup, Ji, Hong, Davidson, Alan J, Horb, Marko E, Miller, Rachel K
• Format: Article
• Language: English
• Subjects: Cell culture; Embryogenesis; Green fluorescent protein; High-throughput screening; Kidney diseases; Kidneys; Nephrons; Secretion; Xenopus; Xenopus laevis
• ISSN: 2073-4425; 2073-4425
• DOI: 10.3390/genes9040197

embryos are an established model for studying kidney development. The nephron structure and genetic pathways that regulate nephrogenesis are conserved between and humans, allowing for the study of human disease-causing genes. 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 :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 :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.