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ZFIN ID: ZDB-FIG-150402-24
Seredick et al., 2014 - Lhx3 and Lhx4 suppress Kolmer-Agduhr interneuron characteristics within zebrafish axial motoneurons. Development (Cambridge, England)   141(20):3900-9 Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Anatomical Terms:
Stage Range: 1-4 somites to Prim-5

Fig. 2

lhx4 and lhx3 are expressed in many cells in the ventral neural tube. All images oriented with anterior to the left. (A,B) Dorsal view of 11hpf embryos. Both lhx3 (A) and lhx4 (B) are expressed beginning at 11hpf in two medial cell stripes. (C,D) Lateral view of spinal cord adjacent to somites 8-12 of 24hpf embryo. Both lhx3 (C) and lhx4 (D) are expressed in the pMN domain; however, lhx4 is expressed in fewer cells than lhx3. (E-H) lhx3 and lhx4 expression in ventral neural tube of Tg(nrp1a:GFP) embryos. (E) At 16hpf, lhx3 and lhx4 are expressed at the dorsoventral level of GFP+ MNs, whereas some GFP+ cells express only lhx3 (arrowheads). (F) At 20hpf, lhx3 and lhx4 expression has expanded to include more dorsal GFP cells (arrowheads). (G,H) At 24hpf and 28hpf, lhx3 and lhx4 are broadly co-expressed in GFP+ and GFP cells throughout the lateral half of ventral spinal cord. (I-L) Lateral views of spinal cord hemisegments adjacent to somites 8-12 co-labeled for Islet and either Lhx3 or Lhx4. Dorsal Islet+ cells are Rohon–Beard sensory neurons. Anti-Islet labels PMNs at 18hpf, and PMNs plus a few SMNs at 24hpf. (I) At 18hpf, anti-Lhx3 labels all Islet+ PMNs as well as slightly more dorsal cells (arrowhead) in ventral neural tube. (J) At 18hpf, anti-Lhx4 labels almost exclusively Islet+ PMNs. (K) By 24hpf, anti-Lhx3 labels all Islet+ PMNs and SMNs and an increased number of cells in ventral neural tube. (L) Anti-Lhx4 also labels all Islet+ PMNs and SMNs in addition to other more dorsal cells (arrowhead) in ventral neural tube at 24hpf. (M) Lateral view of 12hpf Tg(olig2:GFP) embryo co-labeled with anti-Lhx3/4 and anti-phospho-Histone H3 (P-H3). (N) Dorsal view of 12hpf Tg(olig2:GFP) embryo incubated with EdU from 11hpf and labeled with EdU and anti-Lhx3/4. (O) Lateral view of 20hpf Tg(vsx1:GFP) embryo co-labeled with anti-Lhx3/4 and anti-P-H3. Enlarged, single channel images of indicated cell (arrowhead) demonstrate co-expression. (P) Summary of Lhx3- and Lhx4-expressing cells in the ventral spinal cord. (Q) Combinatorial expression of three transcription factors that uniquely label all four neuronal derivatives of the zebrafish pMN domain. Scale bars: 20µm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP vu12Tg standard conditions 5-9 somites posterior neural rod neuroblast IHC
GFP js12Tg standard conditions 14-19 somites posterior neural tube motor neuron IHC
20-25 somites posterior neural tube motor neuron IHC
Prim-5 posterior neural tube motor neuron IHC
nns5Tg standard conditions 20-25 somites posterior neural tube neuroblast IHC
lhx3 WT standard conditions 1-4 somites spinal cord neural keel ventral region ISH
ab1-lhx3 14-19 somites CiD IHC
ab1-lhx3 14-19 somites primary motor neuron IHC
ab1-lhx3 14-19 somites VeLD IHC
ab1-lhx3 Prim-5 CiD IHC
ab1-lhx3 Prim-5 primary motor neuron IHC
ab1-lhx3 Prim-5 secondary motor neuron IHC
Prim-5 spinal cord neural tube ventral region ISH
ab1-lhx3 Prim-5 VeLD IHC
js12Tg standard conditions 14-19 somites posterior neural rod motor neuron ISH
14-19 somites posterior neural rod spinal cord interneuron ISH
20-25 somites posterior neural tube motor neuron ISH
20-25 somites posterior neural tube spinal cord interneuron ISH
Prim-5 spinal cord motor neuron ISH
Prim-5 spinal cord spinal cord interneuron ISH
lhx4 WT standard conditions 1-4 somites spinal cord neural keel ventral region ISH
ab1-lhx4 14-19 somites primary motor neuron IHC
ab1-lhx4 Prim-5 CiD IHC
ab1-lhx4 Prim-5 primary motor neuron IHC
ab1-lhx4 Prim-5 secondary motor neuron IHC
Prim-5 spinal cord neural tube ventral region ISH
ab1-lhx4 Prim-5 VeLD IHC
js12Tg standard conditions 14-19 somites posterior neural rod motor neuron ISH
14-19 somites posterior neural rod spinal cord interneuron ISH
20-25 somites posterior neural tube motor neuron ISH
20-25 somites posterior neural tube spinal cord interneuron ISH
Prim-5 spinal cord motor neuron ISH
Prim-5 spinal cord spinal cord interneuron ISH
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Qualifier Anatomy
ab1-lhx3/4 IHC nns5Tg standard conditions 20-25 somites posterior neural tube neuroblast
IHC vu12Tg standard conditions 5-9 somites Not Detected posterior neural rod neuroblast
Ab2-isl IHC WT standard conditions 14-19 somites primary motor neuron
IHC 14-19 somites Rohon-Beard neuron
IHC Prim-5 Rohon-Beard neuron
IHC Prim-5 secondary motor neuron
Phenotype Details No data available
Acknowledgments:
ZFIN wishes to thank the journal Development (Cambridge, England) for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Development