Background Platelet-derived growth factor receptor β (PDGFRβ) is a tyrosine kinase

Background Platelet-derived growth factor receptor β (PDGFRβ) is a tyrosine kinase receptor known to affect vascular development. is immediately dorsal to the dorsal aorta and potentially regulates blood vessel development in the zebrafish. Using a PDGFR tyrosine kinase inhibitor a morpholino oligonucleotide specific to PDGFRβ and a dominant negative PDGFRβ transgenic line we found that PDGFRβ is necessary for angiogenesis of the intersegmental vessels. Significance/Conclusion Our data provide the first evidence that PDGFRβ signaling is required for zebrafish angiogenesis. We propose a OSU-03012 novel mechanism for zebrafish PDGFRβ signaling that regulates vascular angiogenesis in the absence of mural cells. Introduction The development of blood vessels occurs in two distinct stages. Vasculogenesis is defined as the formation of new blood vessels resulting from angioblast aggregation followed by the lumenization of vascular endothelial cells. Angiogenesis is a secondary process to vasculogenesis and occurs via the sprouting of blood vessels from pre-existing vascular structures [1] [2]. During blood vessel development endothelial cells fuse together to form the vascular lumen initially. Supporting mural cells are then recruited to endothelial cells to stabilize the blood vessel wall and to aid in the formation of the extracellular matrix [3]. The regulation of vasculogenesis and angiogenesis each involves multiple cell types and signaling molecules necessary to coordinate the formation of the vascular system. Platelet-derived growth factor (PDGF) activates a specific family of receptor tyrosine kinases and is involved in the development of blood vessels in chicks and mammals [4]. PDGF-A and PDGF-B ligands can form homo- or heterodimers that bind to the PDGF receptors (PDGFR) α or β. PDGF-B and PDGFRβ null mice die late in embryonic development with renal and cardiovascular abnormalities and fatal hemorrhages [5] [6] [7]. Tissue-specific knockouts show that PDGF-B is secreted by endothelial cells and acts to recruit mural cells (smooth muscle cells OSU-03012 and pericytes) for vascular support [8]. Thus PDGF-B and PDGFRβ paracrine signaling drives the recruitment of smooth FABP4 muscle and pericyte progenitor cells to the wall of new blood vessels [9] [10] [11] [12]. The zebrafish embryo is an excellent model to use for investigating vascular development. The transparency of zebrafish embryos allows for convenient observation during development while transgenic lines that express fluorescent tags in endothelial cells facilitate the study of developing blood vessels [13] [14]. Zebrafish angiogenesis and vasculogenesis are two distinct vascular processes and occur at different phases of vascular development. During zebrafish development the formation of the dorsal aorta and the posterior cardinal vein of the tail occurs via the fusion of angioblast precursor cells and is considered OSU-03012 vasculogenesis. The subsequent sprouting and extension of the intersegmental vessels (ISVs) from the dorsal aorta is considered angiogenesis [15]. More recently it was shown that formation of the posterior cardinal vein occurs via sprouting and segregation from the dorsal aorta in a process that is distinct from vasculogenesis or angiogenesis [16]. Here we provide the first analysis of PDGF signaling in vascular development in the zebrafish. We found that PDGFRβ is expressed adjacent to the dorsal aorta in the hypochord as early as 20 hours post fertilization (hpf). At these early stages of zebrafish development there is no evidence of the presence of mural cells in the vasculature although primitive mural cell markers are present around the anterior portion of the dorsal aorta beginning at 72 hpf [17]. Inhibition of OSU-03012 PDGFR signaling using a PDGFR tyrosine kinase inhibitor caused a decrease in the number and extent of ISV formation. Further morpholino knockdown specific to PDGFRβ and a dominant negative PDGFRβ transgenic line both demonstrated that PDGFRβ is required for angiogenesis of the ISVs. Our results suggest OSU-03012 a new role for PDGFRβ signaling in zebrafish vascular development that functions in the absence of mural cells during early angiogenesis. Results Zebrafish PDGF-B and PDGFRβ were highly conserved with other species We previously cloned and characterized a gene encoding zebrafish PDGF-B [18] and recently cloned the OSU-03012 zebrafish homolog of PDGFRβ2 (Accession No. {“type”:”entrez-nucleotide” attrs :{“text”:”HM439112″.