FIGURE

Fig. 4

ID

ZDB-FIG-190730-23

Publication

Kolvenbach et al., 2019 - Rare Variants in BNC2 Are Implicated in Autosomal-Dominant Congenital Lower Urinary-Tract Obstruction

Other Figures

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Fig. 4

Distal Pronephric-Outlet Obstruction Is Formed by Pronephric Tissue and Causes Dilated Pronephric Ducts and Increased Apoptosis

(A) Tg(HGj4A) GFP-reporter zfl (lateral view) injected with bnc2 MO emphasize the belonging of the distal pronephric-outlet obstruction, visible as a “vesicle” (marked with a white asterisk), to the distal pronephric duct and cloaca. The enlarged inlay shows the same “vesicle” in the sole bright-field image.

(B and C) High-resolution two-photon microscopy dorsal view of the cloacal region (white asterisks) in Tg(HGj4A) zfl injected with Ctrl MO (B) and bnc2 MO (C) indicates the intimate relation of the pronephric-outlet obstruction to the bilateral pronephric ducts. Widths of the pronephric-duct lumen are indicated with white arrows.

(D) Quantification of pronephric-duct widths in Tg(HGj4A) zfl injected with bnc2 MO shows significantly (p = 0.0002, Student’s t test) dilated pronephric ducts compared to controls. PD = pronephric ducts. Data are presented as means with SEM.

(E–G) Cleaved caspase 3 staining in bnc2 MO-injected zfl (56 hpf) shows an increased rate of apoptosis (black arrowheads in G) compared to Ctrl MO-injected embryos (E–F) around the pronephric duct (marked by red lines), the cloacal region, and the central nervous system (not shown). An overview emphasizing and localizing the pronephric regions shown in (F) and (G) is shown in €.

(H and I) Dorsal image from immunofluorescence staining of acetylated tubulin-stained cilia (red, acetTub) and GFP (green) depicts normal cilia morphology in the pronephric ducts in Tg(wt1b:GFP) zfl injected with Ctrl MO (H) and bnc2 MO (I) at 56 hpf. Pronephric ducts are widened in the bnc2 MO zfl as mentioned before. White arrowheads point to glomerular cysts in the bnc2MO zfl. The sole acetylated-tubulin stain (red channel) emphasizes cilia morphology in the respective enlargement inlays of (H) and (I).

Scale bars represent 200 μm (A), 50 μm (magnification of A–C), 100 μm (E–G), 50 μm (H and I), and 25 μm (magnifications of H and I). ^∗∗∗ p < 0.0005.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	AB/TL + CRISPR1-bnc2 AB/TL + CRISPR2-bnc2 AB/TL + CRISPR3-bnc2 AB/TL + MO1-bnc2 li1Tg + MO1-bnc2 nkhgj4aEt + MO1-bnc2
Knockdown Reagents:	CRISPR1-bnc2 CRISPR2-bnc2 CRISPR3-bnc2 MO1-bnc2
Observed In:	brain cloaca apoptotic process post-vent region pronephric duct pronephric duct apoptotic process pronephric duct cilium pronephric duct opening pronephric glomerulus trunk whole organism
Stage Range:	Prim-5 to Protruding-mouth

Phenotype Detail

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Am. J. Hum. Genet.