Impact of hypoxia induced VEGF and its signaling during caudal fin regeneration in Zebrafish

ABSTRACT:Hypoxia is known to play important role during various cellular process, including regeneration. Regeneration is a complex process involving wound healing and tissue repair. We propose that hypoxia might mediate regeneration through angiogenesis involving angiogenic factors such as VEGF, VEGF-R2, NRP1a during the wound healing process. We have chosen Zebrafish model to study the role of hypoxia induced regeneration. Unlike mammals Zebrafish has the ability to regenerate. Hypoxic condition was mimicked using inorganic salt cobalt chloride to study caudal fin regeneration in adult Zebrafish. Intense blood vessel formation, with increased tail fin length experimented at various time points have been observed when adult zebrafish caudal fin partially amputated were exposed to 1% CoCl2. Regeneration is enhanced under hypoxia, with increased VEGF expression. To study the significance of VEGF signaling during wound healing and tissue regeneration, sunitinib well known inhibitor of VEGF receptor is used against CoCl2-induced caudal fin regeneration. Diminished fin length, lowering of blood vessel formation was documented using angioquant software, reduction in mRNA level of hypoxia inducible factors, VEGF and other pro-angiogenic genes such as VEGF, VEGF-R2, NRP1A, FGFR2, ANGPT1 were observed, while reduction in VEGF protein was demonstrated using western blot analysis. Genistein inhibitor of HIF-1α completely arrested regeneration, with suppression of VEGF highlighting the significance of hypoxia induced VEGF signaling during fin regeneration. Our results suggest that hypoxia through HIF-1α might lead to angiogenesis involving VEGF signaling during wound healing and this might throw light on therapeutic efficacy of cobalt chloride during regeneration.

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VEGF-VEGF Receptor 1 (Flt-1) Interactions Significantly Modulate Endothelial Progenitor Cell (EPC) Migration but Have Only Minor Effects on the Migration of Differentiated Endothelial Cells.

Blood ◽

10.1182/blood.v104.11.3229.3229 ◽

2004 ◽

Vol 104 (11) ◽

pp. 3229-3229

Author(s):

Bin Li ◽

Amylynn A. Teleron ◽

Charles Lin ◽

Pampee P. Young

Keyword(s):

Wound Healing ◽

Endothelial Cells ◽

Blood Vessel ◽

Vegf Receptor ◽

Blocking Antibody ◽

Blood Vessel Formation ◽

Migration Assay ◽

Vessel Formation ◽

Cellular Expression

Abstract There is a growing body of data demonstrating that vasculogenesis, whereby bone marrow -derived circulating progenitor cells (EPCs) home to sites of neovascularization, results in significant contribution to blood vessel formation during tumor growth, ischemic injury and wound healing. Vascular Endothelial Growth Factor (VEGF) has recently been shown to augment vasculogenesis. In the current study, we examined if VEGF/VEGF Receptor (VEGFR) interactions are important for EPC recruitment. Both VEGFR1 (flt-1) and VEGFR2 (flk-1) are strongly expressed, as detected by immunofluorescent and FACS analysis, on EPCs obtained by ex vivo expansion of human peripheral blood. In a modified Boyden chamber migration assay, EPCs showed dose dependent migration to VEGF. To examine receptor specificity, EPCs were preincubated with receptor blocking anti-VEGFR1 or -VEGFR2 prior to the migration assay. Level of inhibition by VEGFR1 blocking antibody was commensurate with blocking VEGFR2. Furthermore, migration in response to a VEGFR1-specific agonist, PlGF, was comparable to that induced by VEGF and was completely ablated by preincubation with VEGFR1 blocking antibody. By contrast, differentiated endothelial cells had diminished migration in response to PlGF as compared to VEGF. Furthermore, blocking VEGFR1 only mildly disrupted VEGF-induced migration of differentiated endothelial cells in vitro. Hence, unlike differentiated endothelial cells, EPC migration in vitro was mediated by both VEGF receptors. By quantitative RT-PCR, we examined the level of VEGFR1 and VEGFR2 mRNA transcripts in EPCs versus differentiated endothelial cells. VEGFR1 transcripts in EPCs were expressed 3-fold higher than in differentiated endothelial cells. VEGFR2 and neuropilin transcript levels in EPCs, however, were lower than in differentiated endothelial cells. These results suggest that VEGF/VEGFR1 interactions are important in EPC migration in vitro. We have subcloned VEGFR1 cDNA into a retrovirus vector and have shown by western blot that we can direct increased cellular expression of VEGFR1. In further experiments, we will examine the role of VEGFR1 in human EPC recruitment using murine xenotransplant models of hindlimb ischemia and wound healing. These studies will provide valuable insight towards developing EPCs as cellular therapy to augment blood vessel formation.

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Orientation of endothelial cell division is regulated by VEGF signaling during blood vessel formation

Blood ◽

10.1182/blood-2006-07-037952 ◽

2006 ◽

Vol 109 (4) ◽

pp. 1345-1352 ◽

Cited By ~ 77

Author(s):

Gefei Zeng ◽

Sarah M. Taylor ◽

Janet R. McColm ◽

Nicholas C. Kappas ◽

Joseph B. Kearney ◽

...

Keyword(s):

Cell Division ◽

Endothelial Cell ◽

Blood Vessel ◽

Embryonic Stem ◽

Es Cell ◽

Blood Vessel Formation ◽

Vessel Formation ◽

Vegf Signaling ◽

Vascular Beds

Abstract New blood vessel formation requires the coordination of endothelial cell division and the morphogenetic movements of vessel expansion, but it is not known how this integration occurs. Here, we show that endothelial cells regulate division orientation during the earliest stages of blood vessel formation, in response to morphogenetic cues. In embryonic stem (ES) cell–derived vessels that do not experience flow, the plane of endothelial cytokinesis was oriented perpendicular to the vessel long axis. We also demonstrated regulated cleavage orientation in vivo, in flow-exposed forming retinal vessels. Daughter nuclei moved away from the cleavage plane after division, suggesting that regulation of endothelial division orientation effectively extends vessel length in these developing vascular beds. A gain-of-function mutation in VEGF signaling increased randomization of endothelial division orientation, and this effect was rescued by a transgene, indicating that regulation of division orientation is a novel mechanism whereby VEGF signaling affects vessel morphogenesis. Thus, our findings show that endothelial cell division and morphogenesis are integrated in developing vessels by flow-independent mechanisms that involve VEGF signaling, and this cross talk is likely to be critical to proper vessel morphogenesis.

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Photobiomodulation regulates cytokine release and new blood vessel formation during oral wound healing in rats

Lasers in Medical Science ◽

10.1007/s10103-016-1904-0 ◽

2016 ◽

Vol 31 (4) ◽

pp. 665-671 ◽

Cited By ~ 23

Author(s):

Vivian Petersen Wagner ◽

Marina Curra ◽

Liana Preto Webber ◽

Carolina Nör ◽

Ursula Matte ◽

...

Keyword(s):

Wound Healing ◽

Blood Vessel ◽

Cytokine Release ◽

Blood Vessel Formation ◽

Vessel Formation ◽

Oral Wound Healing

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Faculty Opinions recommendation of Cdc42 is required for cytoskeletal support of endothelial cell adhesion during blood vessel formation in mice.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.725705389.793510324 ◽

2015 ◽

Author(s):

Guillermo Oliver ◽

Noelia Escobedo

Keyword(s):

Cell Adhesion ◽

Endothelial Cell ◽

Blood Vessel ◽

Blood Vessel Formation ◽

Vessel Formation ◽

Endothelial Cell Adhesion

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Emerging Role of AP-1 Transcription Factor JunB in Angiogenesis and Vascular Development

International Journal of Molecular Sciences ◽

10.3390/ijms22062804 ◽

2021 ◽

Vol 22 (6) ◽

pp. 2804

Author(s):

Yasuo Yoshitomi ◽

Takayuki Ikeda ◽

Hidehito Saito-Takatsuji ◽

Hideto Yonekura

Keyword(s):

Endothelial Cells ◽

Transcription Factor ◽

Blood Vessel ◽

Vascular Endothelial Cells ◽

Vascular Development ◽

Endothelial Cell Migration ◽

Vascular Endothelial ◽

Blood Vessel Formation ◽

Vessel Formation

Blood vessels are essential for the formation and maintenance of almost all functional tissues. They play fundamental roles in the supply of oxygen and nutrition, as well as development and morphogenesis. Vascular endothelial cells are the main factor in blood vessel formation. Recently, research findings showed heterogeneity in vascular endothelial cells in different tissue/organs. Endothelial cells alter their gene expressions depending on their cell fate or angiogenic states of vascular development in normal and pathological processes. Studies on gene regulation in endothelial cells demonstrated that the activator protein 1 (AP-1) transcription factors are implicated in angiogenesis and vascular development. In particular, it has been revealed that JunB (a member of the AP-1 transcription factor family) is transiently induced in endothelial cells at the angiogenic frontier and controls them on tip cells specification during vascular development. Moreover, JunB plays a role in tissue-specific vascular maturation processes during neurovascular interaction in mouse embryonic skin and retina vasculatures. Thus, JunB appears to be a new angiogenic factor that induces endothelial cell migration and sprouting particularly in neurovascular interaction during vascular development. In this review, we discuss the recently identified role of JunB in endothelial cells and blood vessel formation.

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