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Figure 3.
Prdm15 is required for glomerular and tubular development in X. laevis. Embryonic stages and scale bars are indicated in each panel and apply to all images included in the corresponding panel. (A) Illustration of the Xenopus pronephros (on the basis of Raciti et al.53 and Cizelsky et al.54) and its corresponding segment-specific marker genes. Overall: fxyd2; glomerulus: wt1; nephrostomes: lhx1; proximal tubule: foxc1 and slc5a1; intermediate tubule: slc12a1; intermediate, distal, and connecting tubule: clcnk; distal and connecting tubule: lhx1. (B) Whole-mount in situ hybridization (WMISH) against prdm15 in X. laevis at stage 33. Specific prdm15 expression was detected in the developing pronephric tissue (arrowhead). Left, lateral view with anterior toward the right; right, close up view of the pronephros. (C) prdm15 is expressed in the X. laevis pronephros at stage 36. Left, sagittal section of WMISH in X. laevis against prdm15 at stage 36 with anterior to the right; right, magnification of the proximal part of the pronephros. Arrowhead points to proximal tubule. (D) fxyd2 at stage 36 shows a specific expression in the pronephric tubule. Left, sagittal section of WMISH in X. laevis against fxyd2 with anterior to the right; right, magnification of the proximal part of the pronephros. Arrowhead points to proximal tubule. (E and F) Human PRDM15 rescues the MO-mediated knockdown of Prdm15. Injection of 15 ng Prdm15 MO leads to the reduction of pronephric-specific marker gene expression of wt1 (Wilms tumor suppressor gene-1) and fxyd2 (-subunit of the Na+/K+-ATPase) (arrowheads), whereas the injection of a control MO has no effect. Lateral views of injected sides with anterior to the right, transversal sections and quantitative representations are given. n, number of independent experiments; N, number of analyzed embryos in total. **P0.01. (G) Quantification of fxyd2 expression in the anterior pronephric area reveals a significant size reduction upon Prdm15 knockdown compared with control MO-injected embryos. Human PRDM15 rescues the renal phenotype. Left, lateral view with anterior toward the right (injected side); middle, appropriate expression area (red); right, quantitative representations. n, number of independent experiments. **P0.01; ****P<0.001. (HL) Prdm15 loss-of-function results in reduced marker gene expression (arrowheads), whereas injection of a control MO has no effect. Lateral views of injected sides with anterior to the right, transversal sections and quantitative representations are given. n, number of independent experiments; N, number of analyzed embryos in total. *P0.05. clcnk, chloride channel protein; foxc1, forkhead box c1; lhx1, lim homeobox 1; slc5a1, solute carrier family 5 member 1; slc12a1, solute carrier family 12 member 1.
Source
Mutations in PRDM15 Are a Novel Cause of Galloway-Mowat Syndrome
Journal of the American Society of Nephrology32(3):580-596, March 2021. |
