ZFIN ID: ZDB-FIG-130210-16
Bercier et al., 2013 - WNK1/HSN2 Mutation in Human Peripheral Neuropathy Deregulates KCC2 Expression and Posterior Lateral Line Development in Zebrafish (Danio rerio). PLoS Genetics   9(1):e1003124 Full text @ PLoS Genet.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: 26+ somites to Day 4
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: 26+ somites

Fig. 5

KCC2 is found in the embryonic zebrafish PLL.

A) Presence of kcc2 in the zebrafish embryo was assessed by in situ hybridization against slc12a5 and reveals staining in the hindbrain (hb) and spinal cord (sc) at 72hpf as well as staining in hair cells of a PLL neuromast (nm) at 4dpf. B) Neuromasts of 4 dpf transgenic Tg(NBT:MAPT-GFP) embryos expressing GFP (green) under neuron-specific beta-tubulin promoter (left upper and middle images) and primordium of a 2dpf transgenic Tg(-8.0cldnb:lynEGFP) embryos expressing GFP under the claudin-b promoter in the membranes of cells composing the primordium (left lower image) were labeled with the Ca2+ indicator Rhod 2-AM (deep red) to show that ionophoresis of glycine failed to evoke Ca2+ transients in 3–4 dpf neuromasts (right black upper traces) but did so in primordium cells (right red bottom traces). In contrast, neuromasts of embryos of equivalent stage respond to glutamate (right, green middle trace). The dashed line illustrates the primordium or neuromast region and the heavy lines illustrates the position of the pipet. C)Transgenic embryos expressing GFP under the claudin-b promoter, which labels the membranes of cells composing the primordium, were used to observe the size of this migrating group of PLL neuromast progenitors for both non-injected and WNK1/HSN2 knockdown embryos. Close-up image of the primordium cells shows no difference in organization between non-injected and WNK1/HSN2 knockdown embryos. D) The primordium area was measured on one side of the embryos and data was tabulated in a scatter plot which shows that WNK1/HSN2 embryos, as well as embryos overexpressing human KCC2, have a significantly smaller primordium area than non-injected embryos. The number of primordial measured is indicated in parenthesis at the bottom of the graph. The knockdown embryo presented in (C) is a representative results at 22hpf obtained from MO-wnk1-ATG injection. Scale bars: (A) 100μm and 20μm for neuromast image, (B)20μm, (C) 80μm for full primordium and 20μm for cell close-up.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP zf106Tg standard conditions 26+ somites posterior lateral line primordium plasma membrane IFL
zf106Tg + MO2-wnk1b standard conditions 26+ somites posterior lateral line primordium plasma membrane IFL
slc12a5b WT standard conditions Protruding-mouth hindbrain ISH
Protruding-mouth spinal cord anterior region ISH
Day 4 posterior lateral line neuromast neuromast hair cell ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
zf106Tg + MO1-wnk1b standard conditions 26+ somites posterior lateral line development decreased process quality, abnormal
26+ somites posterior lateral line primordium decreased area, abnormal
zf106Tg + MO2-wnk1b standard conditions 26+ somites posterior lateral line development decreased process quality, abnormal
26+ somites posterior lateral line primordium decreased area, abnormal
zf106Tg + MO3-wnk1b standard conditions 26+ somites posterior lateral line development decreased process quality, abnormal
26+ somites posterior lateral line primordium decreased area, abnormal
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 @ PLoS Genet.