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FIGURE 2

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Figures for Martin et al., 2022
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FIGURE 2

HCR RNA-FISH validates bioinformatic findings and spatially locates tectal populations. Hybridization chain reaction RNA-fluorescence in situ hybridization (HCR RNA-FISH) was performed using proprietary custom probe sets designed by Molecular Instruments to spatially map populations and validate bioinformatic exclusion of non-interest cells. (A–A”,C–C”,E–E”) Single z-slices of representative images showing expression of habenula (gng8) and OT (nrxn3a, robo4) markers in 7 dpf larval zebrafish. (A–A”) gng8+ cells are located in the habenula, validating gng8 expression as a method for annotating habenular cells and spatially mapping 89% of habenula cells (Supplementary Table 2). (B) Feature plot showing gng8 expression is restricted to the annotated habenula cluster. (C–C”) nrxn3a+ cells are primarily located medially within the periventricular layer, and spatially map 54% of OT5 (Supplementary Table 2). (D) Feature plot showing expression of nrxn3a is enriched in OT5 and to a lesser extent, OT8. (E–E”) robo4+ OT cells are located near the intratectal commissure, likely within the tectal proliferation zone, and spatially map 51% of OT2 cells (Supplementary Table 2). (F) Feature plot showing robo4 expression is restricted to OT2. All HCR RNA-FISH panels include DAPI staining as a cellular reference.

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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 @ Front. Mol. Neurosci.