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ZFIN ID: ZDB-FIG-090504-14
Zeng et al., 2009 - Phospholipase D1 is required for angiogenesis of intersegmental blood vessels in zebrafish. Developmental Biology   328(2):363-376 Full text @ Dev. Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Condition:
Knockdown Reagents:
Anatomical Terms:
Stage Range: Prim-25 to Long-pec
PHENOTYPE:
Fish:
Condition:
Knockdown Reagents:
Observed In:
Stage Range: Prim-25 to Long-pec

Fig. 7 (A,B) Disruption of Pld1 signaling differentially affects ISV and motor axon dorsal–ventral morphogenesis. (A) Uninjected control TG(flk1:GFP) embryos were stained with znp1 antibody to visualize primary motor neurons (red), which extend from the dorsal to the ventral side along with ISV (green). (B) Anti-acetylated tubulin antibody detects both the primary and secondary motor neuron cell body and axons (red) in the trunk region of uninjected control TG(fli1:GFP) embryos, and ISV (green) is positioned in between. Embryos with MO1-pld1 injection or 1-butanol treatment exhibit impaired ISV morphology compared with control embryos; however, all the motor neurons are able to connect from the ventral side to the dorsal side. Interestingly, the motoneurons are more branched when Pld1 signaling is disrupted (white arrowheads), compared to control embryos. (C–E,H) Transplanted wild-type cells in the notochord partially rescue the ISV defects in pld1 morphants. (C–E) Fluorescent images of TG(fli:GFP) pld1 morphants containing Rhodamine-dextran labeled cells (red) transplanted from wild-type donor embryos, and localized in the prechordal plate (C) or anterior notochord (D,E). Lateral view, anterior to the left. ISVs are visualized in green by TG(fli:GFP). White arrows point to normal ISVs in the regions of the chimeric embryos possessing transplanted wild-type cells, whereas red arrows indicate defective ISVs in segments lacking wild-type cells. (H) In chimeric embryos harboring transplanted cells in the notochord, the percentage of ISV defects in segments with wild-type cells (12%, segments number = 34, 8 embryos) compared to segments without wild-type cells (33%, segments number = 164, 15 embryos). (F,G,I) Transplanted wild-type cells in the somites cannot rescue the ISV defects in pld1 morphants. (F,G) Fluorescent images of TG(fli:GFP) pld1 morphants containing Rhodamine-dextran labeled cells (red) transplanted from wild-type donor embryos, and localized in the somites. Lateral view, anterior to the left. ISVs are visualized in green by TG(fli:GFP). White arrows with red filling point to defective ISVs in the regions of the chimeric embryos possessing transplanted wild-type cells, whereas red arrows with white filling indicate defective ISVs in segments lacking wild-type cells. (I) In the chimeras exhibiting transplanted cells in the somites, the percentage of ISV defects in segments with wild-type cells (51%, segments number = 124, 9 embryos) was comparable to segments without wild-type cells (43%, segments number = 92, 9 embryos). Scale bar represents 20 μm (A) and 30 μm (C–G).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP y1Tg standard conditions Long-pec intersegmental vessel IFL
y1Tg chemical treatment: butan-1-ol Long-pec intersegmental vessel IFL
y1Tg + MO2-pld1a standard conditions Long-pec intersegmental vessel IFL
grcfp zn1Tg standard conditions Prim-25 intersegmental vessel IFL
zn1Tg chemical treatment: butan-1-ol Prim-25 intersegmental vessel IFL
zn1Tg + MO1-pld1a standard conditions Prim-25 intersegmental vessel IFL
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
znp-1 IHC zn1Tg standard conditions Prim-25 primary motor neuron
IHC zn1Tg chemical treatment: butan-1-ol Prim-25 primary motor neuron
IHC zn1Tg + MO1-pld1a standard conditions Prim-25 primary motor neuron
Phenotype Details
Fish Conditions Stage Phenotype
y1Tg chemical treatment: butan-1-ol Long-pec angiogenesis disrupted, abnormal
Long-pec intersegmental vessel morphology, abnormal
Long-pec motor neuron axon guidance process quality, normal
Long-pec primary motor neuron branched, abnormal
y1Tg + MO2-pld1a standard conditions Long-pec angiogenesis disrupted, abnormal
Long-pec intersegmental vessel morphology, abnormal
Long-pec motor neuron axon guidance process quality, normal
Long-pec primary motor neuron branched, abnormal
zn1Tg chemical treatment: butan-1-ol Prim-25 angiogenesis disrupted, abnormal
Prim-25 intersegmental vessel morphology, abnormal
Prim-25 motor neuron axon guidance process quality, normal
zn1Tg + MO1-pld1a standard conditions Prim-25 angiogenesis disrupted, abnormal
Prim-25 intersegmental vessel morphology, abnormal
Prim-25 motor neuron axon guidance process quality, normal
Prim-25 primary motor neuron branched, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Biology for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Developmental Biology, 328(2), Zeng, X.X., Zheng, X., Xiang, Y., Cho, H.P., Jessen, J.R., Zhong, T.P., Solnica-Krezel, L., and Brown, H.A., Phospholipase D1 is required for angiogenesis of intersegmental blood vessels in zebrafish, 363-376, Copyright (2009) with permission from Elsevier. Full text @ Dev. Biol.