ZFIN ID: ZDB-FIG-140515-17
van Impel et al., 2014 - Divergence of zebrafish and mouse lymphatic cell fate specification pathways. Development (Cambridge, England)   141(6):1228-1238 Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Anatomical Terms:
Stage Range: Prim-15 to Day 5
PHENOTYPE:
Fish:
Observed In:
Stage Range: Long-pec to Day 5

Fig. 2

Lymphangiogenesis in prox1a mutant embryos. (A) Schematic of the homeodomain (HD) containing Prox1a protein, indicating the predicted effect of the 10 bp deletion in the prox1ai278 allele, leading to a frame-shift (red amino acids) and a truncated protein after 153 amino acids. (B,C) Prox1a immunostaining of slow muscle fibers in sibling (B) and homozygous mutant prox1ai278 (C) embryos demonstrates a complete loss of wild-type Prox1a protein (green) at 30 hpf (slow myosin heavy chain-1 is shown in red). (D,E) Brightfield pictures of 5 dpf sibling (D) and homozygous prox1ai278 mutant (E) embryos. Note the strong edema formation around the eye and gut area (arrowheads), which can be even more pronounced in other prox1a mutants at this stage. (F,G) In both heterozygous siblings (F) and homozygous prox1ai278 mutants (G), PLs appear at the level of the horizontal myoseptum at 2 dpf (arrows). (H) Average PL numbers per embryo are mildly reduced in prox1a mutants at 2 dpf (Student’s t-test, *P=0.025). Error bars indicate s.d. of wild-type (green), heterozygous (orange) and mutant (red) groups in embryos from a prox1a+/- incross. (I-K) flt4:mCit; flt1enh:tdTom double transgenic embryos highlighting arterial ISVs in red and venous and lymphatic structures in green. Compared with heterozygous siblings (I), most homozygous prox1ai278 mutants do not display TD defects at 5 dpf (J), whereas others display a mild reduction (K) in some areas of the trunk (arrows point at TD; asterisks mark the lack of TD). Note the overall unaffected ratio of venous and arterial ISVs in mutants (J,K). (L) Average number of segments positive for TD cells, scored in the first ten segments above the yolk extension at 5 dpf. Error bars indicate the s.d. for the respective genotypic class from a prox1a+/- incross. ***P=2.3E-08 (Student’s t-test, comparison of wild-type and mutant population). (M,N) The average percentage of intersegmental veins (M) and arteries (N) does not differ between genotypic classes in an incross of prox1ai278 carriers. (O) In prox1ai278 mutants, the average number of ISVs is not altered. Error bars represent s.d. n.s., not statistically significant.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
Citrine hu5333Tg; hu7135Tg standard conditions Long-pec lymphatic system IFL
Long-pec vascular lymphangioblast IFL
Long-pec vein IFL
Day 5 intersegmental lymph vessel IFL
Day 5 intersegmental vein IFL
Day 5 lymph vasculature IFL
Day 5 thoracic duct IFL
Day 5 vein IFL
prox1ai278/i278; hu5333Tg; hu7135Tg standard conditions Long-pec lymphatic system IFL
Long-pec vascular lymphangioblast IFL
Long-pec vein IFL
Day 5 intersegmental lymph vessel IFL
Day 5 intersegmental vein IFL
Day 5 lymph vasculature IFL
Day 5 thoracic duct IFL
Day 5 vein IFL
RFP hu5333Tg; hu7135Tg standard conditions Long-pec intersegmental vessel artery IFL
Day 5 intersegmental vessel artery IFL
prox1ai278/i278; hu5333Tg; hu7135Tg standard conditions Long-pec intersegmental vessel artery IFL
Day 5 intersegmental vessel artery IFL
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Qualifier Anatomy
Ab-F59 IHC WT standard conditions Prim-15 slow muscle cell
IHC prox1ai278/i278 standard conditions Prim-15 slow muscle cell
Ab3-prox1 IHC WT standard conditions Prim-15 myotome
IHC prox1ai278/i278 standard conditions Prim-15 Not Detected myotome
Phenotype Details
Fish Conditions Stage Phenotype
prox1ai278/i278 standard conditions Day 5 whole organism edematous, abnormal
prox1ai278/i278; hu5333Tg; hu7135Tg standard conditions Long-pec vascular lymphangioblast decreased amount, abnormal
Day 5 thoracic duct decreased length, abnormal
Day 5 trunk anterior region lacks all parts of type thoracic duct, abnormal
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