Maternal and Zygotic Sphingosine Kinase 2 Are Indispensable for Cardiac Development in Zebrafish

Hisano, Yu; Inoue, Asuka; Okudaira, Michiyo; Taimatsu, Kiyohito; Matsumoto, Hirotaka; Kotani, Hirohito; Ohga, Rie; Aoki, Junken; Kawahara, Atsuo

doi:10.1074/jbc.m114.634717

Cited by 16 publications

(15 citation statements)

References 45 publications

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“…Sphk1 morpholino KD produced very few effects on the global phenotype, as observed in previous studies 16,17 . However, Sphk1 KD impaired ISV formation, with a phenotype similar, although less severe, compared to that observed following Lmo2 KD.…”

Section: Discussionsupporting

confidence: 85%

“…4A). There is only one group that has examined Sphk in zebrafish, finding that Sphk2, but not Sphk 1, 16,17 mediates cardiac development. Accordingly, it was necessary for us to clone and sequence Sphk1 cDNA before embarking upon Sphk1 loss-of-function studies in vivo .…”

Section: Resultsmentioning

confidence: 99%

“…Knockout of either Sphk1 or Sphk2 generates phenotypically normal mice or zebrafish 16,17 , suggesting that Sphk1 and Sphk2 have redundant functions that can compensate for each other. However recent studies revealed that overexpression of Sphk2 suppressed cell growth and enhanced apoptosis in vitro 30 , in contrast to Sphk1 , which generally promoted cell survival and growth 16,17 . The contrasting effects of their overexpression may be due to their different cellular localization, as Sphk1 in human is mostly localized in the cytosol whereas Sphk2 localizes to the mitochondria, endoplasmic reticulum and nucleus 27 .…”

Section: Discussionmentioning

confidence: 99%

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Lmo2 (LIM-Domain-Only 2) Modulates Sphk1 (Sphingosine Kinase) and Promotes Endothelial Cell Migration

Matrone

Meng

et al. 2017

ATVB

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Objective Lim-domain only (Lmo)2 transcription factor is involved in hematopoiesis and vascular remodeling. Sphingosine kinase (Sphk)1 phosphorylates sphingosine to sphingosine-1-phosphate (S1P). We hypothesized that Lmo2 regulates Sphk1 to promote endothelial cell (EC) migration and vascular development. Approach and Results Lmo2 and Sphk1 knockdown (KD) were performed in Tg(fli1:EGFP)y1 zebrafish and in HUVEC. Rescue of phenotypes or overexpression of these factors were achieved using mRNA encoding Lmo2 or Sphk1. Endothelial cell proliferation in vivo was assessed by BrdU immunostaining and FACS analysis of dissociated Tg(fli1:EGFP)y1 embryos. Cell migration was assessed by scratch assay in HUVEC and mouse aortic rings. Lmo2 interactions with Sphk1 promoter were assessed by ChIP-PCR. Lmo2 or Sphk1 KD reduced number and length of intersegmental vessels (ISVs). There was no reduction in the numbers of GFP+ endothelial cells following Lmo2 KD. However, reduced numbers of BrdU+GFP+ nuclei were observed along the dysmorphic ISVs, accumulating instead at the sprouting origin of the ISVs. This anomaly was likely due to impaired EC migration, which was confirmed in migration assays using Lmo2 KD HUVECs and mouse aortic rings. Both in vivo and in vitro, Lmo2 KD reduced Sphk1 gene expression, associated with less Lmo2 binding to the Sphk1 promoter as assessed by ChIP-PCR. Sphk1 mRNA rescued the Lmo2 KD phenotype. Conclusions Our data showed that Lmo2 is necessary for Sphk1 gene expression in endothelial cells. Lmo2 KD reduced Lmo2-Sphk1 gene interaction, impaired ISVs formation and reduced cell migration. We identified for the first time Sphk1 as downstream effector of Lmo2.

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Section: Discussionsupporting

confidence: 85%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Lmo2 (LIM-Domain-Only 2) Modulates Sphk1 (Sphingosine Kinase) and Promotes Endothelial Cell Migration

Matrone

Meng

et al. 2017

ATVB

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“…; Hisano et al . , ). Another possibility is that each s1pr in zebrafish redundantly functions in a unique fashion.…”

Section: Discussionmentioning

confidence: 99%

Comprehensive analysis of sphingosine‐1‐phosphate receptor mutants during zebrafish embryogenesis

et al. 2015

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The lipid mediator sphingosine-1-phosphate (S1P) regulates various physiological and pathological phenomena such as angiogenesis and oncogenesis. Secreted S1P associates with the G-protein-coupled S1P receptors (S1PRs), leading to the activation of downstream signaling molecules. In mammals, five S1prs have been identified and the genetic disruption of a single S1pr1 gene causes vascular defects. In zebrafish, seven s1prs have been isolated. We found that individual s1prs showed unique expression patterns with some overlapping expression domains during early embryogenesis. We generated all s1pr single-mutant zebrafish by introducing premature stop codons in their coding regions using transcription activator-like effector nucleases and analyzed their phenotypes during early embryogenesis. Zygotic s1pr1, s1pr3a, s1pr3b, s1pr4, s1pr5a and s1pr5b mutants showed no developmental defects and grew into adults, whereas zygotic s1pr2 mutant showed embryonic lethality with a cardiac defect, showing quite distinct embryonic phenotypes for individual S1pr mutants between zebrafish and mouse. We further generated maternal-zygotic s1pr1, s1pr3a, s1pr3b, s1pr4, s1pr5a and s1pr5b mutants and found that these maternal-zygotic mutants also showed no obvious developmental defects, presumably suggesting the redundant functions of the S1P receptor-mediated signaling in zebrafish.

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“…The plasmids used to synthesize TALEN mRNAs were constructed as described previously ( Hisano et al, 2015 ). RVD repeat arrays were cloned into pCS2TAL3DD and pCS2TAL3RR to generate left and right TALEN constructs ( mib2 -TALEN-F and mib2 -TALEN-R).…”

Section: Methodsmentioning

confidence: 99%

Mindbomb 2 is dispensable for embryonic development and Notch signalling in zebrafish

et al. 2015

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The Mindbomb E3 ubiquitin protein ligase (Mib) family of proteins, Mib1 and Mib2, are RING finger ubiquitin ligases that share specific substrates. Mib1 is known to play essential roles in Notch signalling by ubiquitinating Notch ligands in vivo. Conversely, the functions of Mib2 in vivo are not fully understood, although Mib2 ubiquitinates multiple substrates, including Notch ligands, in vitro. To determine the Notch-dependent and Notch-independent functions of Mib2 in vivo, we generated mutant alleles of zebrafish mib2 using transcription activator-like effector nucleases (TALENs). We found that mib2 homozygous mutants were viable and fertile. Notch-mediated functions, such as early neurogenesis, somitogenesis, and pigment cell development, were not affected in mib2 mutant embryos. The lack of Notch-deficient phenotypes in mib2 mutants was not due to compensation by a mib2 maternal gene product because mib2 maternal-zygotic mutants also did not exhibit a distinct phenotype. We also showed that Mib2 does not redundantly act with Mib1 because the genetic ablation of mib2 neither enhanced mibtfi91-null phenotypes nor did it alleviate antimorphic mibta52b phenotypes. Furthermore, the postulated Notch-independent roles of Mib2 in maintaining muscular integrity and N-methyl-D-aspartate receptor (NMDAR) activity were not evident: mib2 mutants did not show phenotypes different from that of the control embryos. These observations suggest that Mib2 is dispensable for embryonic development and does not have redundant functions with Mib1 in Notch signalling at least during early development stages in zebrafish.

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Maternal and Zygotic Sphingosine Kinase 2 Are Indispensable for Cardiac Development in Zebrafish

Cited by 16 publications

References 45 publications

Lmo2 (LIM-Domain-Only 2) Modulates Sphk1 (Sphingosine Kinase) and Promotes Endothelial Cell Migration

Lmo2 (LIM-Domain-Only 2) Modulates Sphk1 (Sphingosine Kinase) and Promotes Endothelial Cell Migration

Comprehensive analysis of sphingosine‐1‐phosphate receptor mutants during zebrafish embryogenesis

Mindbomb 2 is dispensable for embryonic development and Notch signalling in zebrafish

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