FIGURE

Fig. S2

ID

ZDB-FIG-180104-7

Publication

Djenoune et al., 2017 - The dual developmental origin of spinal cerebrospinal fluid-contacting neurons gives rise to distinct functional subtypes

Other Figures

All Figure Page

Back to All Figure Page

Fig. S2

Use of the APEX2 peroxidase combined with the UAS-Gal4 system and pkd2l1 specificity to target CSF-cNs.

(a) Schematic showing how APEX2 is used to generate diaminobenzidine (DAB) contrast for EM in zebrafish CSF-cNs taking advantage of the UAS-Gal4 system. We injected the (UAS:APEX2- TagRFP) construct into Tg(pkd2l1:Gal4)icm10 fertilized eggs in order to get specific expression of APEX2 in CSF-cNs. After screening for TagRFP expression, larvae were fixed and bathed with a solution of DAB and H2O2. APEX2 catalyzes the polymerization and local deposition of DAB, which subsequently attracts electron-dense osmium (OsO4), giving EM contrast only in APEX2 expressing structures namely CSF-cNs (ventral and dorsal). (b1) Lateral view of a Tg(pkd2l1:Gal4)icm10 larva at 2.5 dpf injected with (UAS:APEX2-TagRFP) showing that pkd2l1 promoter drives APEX2-TagRFP expression in CSF-cNs within the spinal cord. (b2) Close up of the rostral spinal cord from a lateral view in the same transgenic animal showing the TagRFP expression in CSF-cNs. (c) Co-injection of (pkd2l1:Gal4) and (UAS:APEX2-TagRFP) within Tg(pkd2l1:GCaMP5G)icm07 embryos where all CSF-cNs are labeled (green) demonstrates the variegated expression of TagRFP within CSF-cNs (magenta). Note that in this panel, the expression of TagRFP is restricted in some dorsal CSF-cNs. Scale bars: 300 μm (b1) and 20 μm (b2, c).

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

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 @ Sci. Rep.