ZFIN ID: ZDB-FIG-151214-75
Barreiro-Iglesias et al., 2015 - Serotonin Promotes Development and Regeneration of Spinal Motor Neurons in Zebrafish. Cell Reports   13:924-932 Full text @ Cell Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Condition:
Knockdown Reagent:
Anatomical Terms:
Stage: Prim-15
PHENOTYPE:
Fish:
Condition:
Knockdown Reagent:
Observed In:
Stage: Prim-15

Fig. 1

Serotonin Signaling Promotes Embryonic Motor Neuron Generation

Lateral views of spinal cords at 33 hpf are shown.

(A–F) Serotonin (5-HT) treatment (24–33 hpf) increases the number of HB9 immuno-labeled motor neurons but has no influence on vsx1:GFP (A–C) and pax2a:GFP labeled interneurons (D–F) in the same embryos (Student’s t test in C, p = 0.0077; in F, p = 0.002).

(G–I) Serotonin treatment increases the number of dividing (pH3+) olig2:GFP+ pMN progenitor cells (Student’s t test in I; p = 0.0006).

(J–L) Lateral view of a double-transgenic olig2:dsRed/HB9:GFP embryo is shown with red only (arrows, pMN progenitors) and double-labeled (arrowheads, motor neurons) cells indicated in the spinal cord (J). A typical FACS profile is shown (K). In RT-PCR, serotonin receptors show enrichment in pMN progenitor cells, compared to motor neurons (L). GAPDH is used for comparison.

(M–O) Morpholino knockdown of receptor htr1ab reduces the number of HB9+ motor neurons but does not influence the number of vsx1:GFP+ interneurons in the same embryos (Student’s t test; p < 0.0001).

The scale bar in (B) represents 10 µm for (A) and (B), in (E) represents 10 µm for (D) and (E), in (H) represents 15 µm for (G) and (H), and in (N) represents 15 µm for (M) and (N). See also Figure S1.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
DsRed2 ml2Tg; vu19Tg standard conditions Prim-15 spinal cord neuroblast IFL
EGFP zf532Tg control Prim-15 spinal cord neuroblast IHC
zf532Tg chemical treatment by environment: serotonin Prim-15 spinal cord neuroblast IHC
GFP e1Tg control Prim-15 spinal cord interneuron IHC
e1Tg chemical treatment by environment: serotonin Prim-15 spinal cord interneuron IHC
ml2Tg; vu19Tg standard conditions Prim-15 spinal cord motor neuron IFL
nns5Tg control Prim-15 spinal cord interneuron IHC
nns5Tg chemical treatment by environment: serotonin Prim-15 spinal cord interneuron IHC
nns5Tg + MO1-htr1ab standard conditions Prim-15 spinal cord interneuron IHC
htr1aa ml2Tg; vu19Tg standard conditions Prim-15 spinal cord neuroblast RTPCR
htr1ab ml2Tg; vu19Tg standard conditions Prim-15 spinal cord motor neuron RTPCR
Prim-15 spinal cord neuroblast RTPCR
htr1b ml2Tg; vu19Tg standard conditions Prim-15 spinal cord motor neuron RTPCR
htr1d ml2Tg; vu19Tg standard conditions Prim-15 spinal cord motor neuron RTPCR
Prim-15 spinal cord neuroblast RTPCR
htr1e ml2Tg; vu19Tg standard conditions Prim-15 spinal cord motor neuron RTPCR
Prim-15 spinal cord neuroblast RTPCR
htr2aa ml2Tg; vu19Tg standard conditions Prim-15 spinal cord motor neuron RTPCR
Prim-15 spinal cord neuroblast RTPCR
htr3a ml2Tg; vu19Tg standard conditions Prim-15 spinal cord neuroblast RTPCR
htr4 ml2Tg; vu19Tg standard conditions Prim-15 spinal cord neuroblast RTPCR
htr5aa ml2Tg; vu19Tg standard conditions Prim-15 spinal cord motor neuron RTPCR
Prim-15 spinal cord neuroblast RTPCR
htr7al ml2Tg; vu19Tg standard conditions Prim-15 spinal cord neuroblast RTPCR
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab1-mnx1 IHC e1Tg control Prim-15 spinal cord motor neuron
IHC e1Tg chemical treatment by environment: serotonin Prim-15 spinal cord motor neuron
IHC nns5Tg control Prim-15 spinal cord motor neuron
IHC nns5Tg chemical treatment by environment: serotonin Prim-15 spinal cord motor neuron
IHC nns5Tg + MO1-htr1ab standard conditions Prim-15 spinal cord motor neuron
IHC zf532Tg control Prim-15 spinal cord motor neuron
Phenotype Details
Fish Conditions Stage Phenotype
e1Tg chemical treatment by environment: serotonin Prim-15 spinal cord motor neuron ab1-mnx1 labeling increased amount, abnormal
nns5Tg chemical treatment by environment: serotonin Prim-15 spinal cord motor neuron ab1-mnx1 labeling increased amount, abnormal
nns5Tg + MO1-htr1ab standard conditions Prim-15 spinal cord interneuron normal amount, normal
Prim-15 spinal cord motor neuron ab1-mnx1 labeling decreased amount, abnormal
Prim-15 spinal cord motor neuron decreased amount, abnormal
Prim-15 spinal cord motor neuron differentiation disrupted, abnormal
zf532Tg chemical treatment by environment: serotonin Prim-15 spinal cord motor neuron differentiation increased process quality, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Cell Reports for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Cell Rep.