ZFIN ID: ZDB-FIG-210708-24
Mikdache et al., 2021 - Rgs4 is a regulator of mTOR activity required for motoneuron axon outgrowth and neuronal development in zebrafish. Scientific Reports   11:13338 Full text @ Sci. Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Antibodies:
Fish:
Anatomical Terms:
Stage Range: 26+ somites to Long-pec
PHENOTYPE:
Fish:
Observed In:
Stage Range: 26+ somites to Long-pec

Figure 2

MZrgs4 embryos show motility and motoneurons axon outgrowth defects. (A) Quantification of the percentage of embryos that showed a normal response to the touch test at 48 hpf in WT (average of 94.44 ± 5.556, n = 18) and MZrgs4 embryos (average of 30.77 ± 7.48, n = 39). (B,C) Z projections of confocal microscopy images of HuC:gfp (B) and HuC:gfp/MZrgs4 mutant (C) showing the extending axons of primary motoneurons at 22 hpf. Arrows indicate the tip ends of extending axons. Scale bar = 20 μm. (D) Quantification of the motor axons length at 22 hpf in HuC:gfp (average of 30.20 ± 1.78 μm, 25 motor axons, n = 9) and MZrgs4/HuC:gfp (average of 31.14 ± 1.01 μm, 36 motor axons, n = 12). (EH) Z Projections of whole-mount immunostaining for Znp1 showing the extending axons of primary motoneurons at 30 hpf in WT (E) and MZrgs4 (F) and the ramifications or branching of these motoneurons at 48 hpf in WT (G) and MZrgs4 embryos (H) Scale bar = 25 μm. (I) Quantification of the motor axons length at 30 hpf in WT (average of 106.7 ± 2.44 μm, 42 motor axons, n = 12) and MZrgs4 embryos (average of 64.37 ± 6.66 μm, 50 motor axons, n = 12). (J) Quantification of the number of ramifications per motoneuron at 48 hpf in WT (average of 20.64 ± 1.10, 36 motoneurons, n = 11) and MZrgs4 embryos (average of 9.37 ± 0.86, 53 motoneurons, n = 16). (K) Acetylated tubulin expression in WT and MZrgs4 embryo at 48 hpf showing the PLLn nerve (white arrows). Scale bar = 50 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP knu3Tg standard conditions 26+ somites motor neuron axon IFL
26+ somites motor neuron cell body IFL
rgs4kbc2/kbc2; knu3Tg standard conditions 26+ somites motor neuron axon IFL
26+ somites motor neuron cell body IFL
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-tuba IHC WT standard conditions Long-pec posterior lateral line nerve axon
IHC rgs4kbc2/kbc2 standard conditions Long-pec posterior lateral line nerve axon
znp-1 IHC WT standard conditions Prim-15 CaP motoneuron axon
IHC Prim-15 MiP motor neuron axon
IHC Prim-15 RoP motor neuron axon
IHC Long-pec CaP motoneuron axon
IHC Long-pec MiP motor neuron axon
IHC Long-pec RoP motor neuron axon
IHC rgs4kbc2/kbc2 standard conditions Prim-15 CaP motoneuron axon
IHC Prim-15 MiP motor neuron axon
IHC Prim-15 RoP motor neuron axon
IHC Long-pec CaP motoneuron axon
IHC Long-pec MiP motor neuron axon
IHC Long-pec RoP motor neuron axon
Phenotype Details
Fish Conditions Stage Phenotype
rgs4kbc2/kbc2 standard conditions Prim-15 CaP motoneuron axon decreased length, abnormal
Prim-15 CaP motoneuron axon extension decreased process quality, abnormal
Long-pec CaP motoneuron axon decreased branchiness, abnormal
Long-pec CaP motoneuron axon decreased length, abnormal
Long-pec CaP motoneuron axon extension decreased process quality, abnormal
Long-pec posterior lateral line nerve axon extension normal process quality, normal
Long-pec posterior lateral line nerve morphology, normal
Long-pec thigmotaxis decreased process quality, abnormal
rgs4kbc2/kbc2; knu3Tg standard conditions 26+ somites primary motor neuron axon extension normal process quality, normal
26+ somites primary motor neuron axon normal length, normal
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.