Development Of Vascular System Research Articles

Ocular vessel patterning in zebrafish is indirectly regulated by Hedgehog signaling.

The superficial ocular vasculature of the embryonic zebrafish develops in a highly stereotypic manner and hence provides a convenient model for studying molecular mechanisms that regulate vascular patterning. We have used transgenic zebrafish embryos in which all endothelial cells express enhanced Green Fluorescent Protein and small molecule inhibitors to examine the contribution of two signaling pathways, vascular endothelial growth factor (VEGF) and Hedgehog (Hh) pathways, to the development of the superficial system. We find that most, but not all vessels of the superficial system depend on VEGF signaling for their growth. Hh signaling appears to limit superficial vessel growth over the dorsal eye and is required to promote superficial vessel growth over the ventral eye. These effects of Hh signaling are indirect. Our initial analyses of factors that regulate growth and patterning of superficial ocular vessels suggest that early patterning events in the embryo during organogenesis stages could influence vascular patterning later on. By studying development of specific vascular systems it should be possible to identify new roles for signaling pathways in regulating vascular development.

Connective Tissue Growth Factor Transgenic Mouse Develops Cardiac Hypertrophy, Lean Body Mass and Alopecia.

Connective Tissue Growth Factor (CTGF/CCN2) is one of the six members of cysteine-rich, heparin-binding proteins, secreted as modular protein and recognised to play a major function in cell processes such as adhesion, migration, proliferation and differentiation as well as chondrogenesis, skeletogenesis, angiogenesis and wound healing. The capacity of CTGF to interact with different growth factors lends an important role during early and late development, especially in the anterior region of the embryo. CTGF Knockout (KO) mice have several craniofacial defects and bone miss shaped due to an impairment of the vascular system development during chondrogenesis. The aim of the study was to establish an association between multiple modular functions of CTGF and the phenotype and cardiovascular functions in transgenic mouse. Bicistronic cassette was constructed using pIRES expressing vector (Clontech, Palo Alto, CA). The construct harbours mouse cDNA in tandem with LacZ cDNA as a reporter gene under the control of Cytomegalovirus (CMV) promoter. The plasmid was linearised with NotI restriction enzyme, and 50 ng of linearised plasmid was injected into mouse pronucleus for the chimaera production. Immunohistochemical methods were used to assess the colocalisation renin and CTGF as well as morphology and rheology of the cardiovascular system. The chimeric mice were backcrossed against the wild-type C57BL/6 to generate hemizygous (F1) mouse. Most of the offsprings died as a result of respiratory distress and those that survived have low CTGF gene copy number, approximately 40 molecules per mouse genome. The copy number assessment on the dead pups showed 5×103 molecules per mouse genome explaining the threshold of the gene in terms of toxicity. Interestingly, the result of this cross showed 85% of the progenies to be positive deviating from Mendelian first law. All F2 progenies died excluding the possibility of establishing the CTGF transgenic mouse line, situation that compelled us to work at the level of hemizygosity. The histological characterisation of left ventricle shows cardiac hypertrophy together with decrease in body mass and alopecia, this compared to the wild type. The immunohistochemical staining of aorta root showed hyperplasia with increased expression and colocalisation of renin and CTGF demonstrating that CTGF may be involved in vascular tone control. Genetic engineering is a noble avenue to investigate the function of new or existing genes. Our data have shown that CTGF transgenic mouse has cardiac and aorta root hypertrophy and abnormal renin accumulation in aorta root as compared to the wild-type animals. The transgenic animals developed alopecia and lean body mass adding two new functions on pre-existing CTGF multiple functions.

Hypoxia inhibits lymphatic thoracic duct formation in zebrafish

Hypoxia promotes blood vessel growth through up-regulation of pro-angiogenic pathways but its role on the lymphatic system remains unclear. The homeobox transcription factor Prox1 is a master control gene for generating lymphatic endothelial cells (LECs) and is up-regulated by hypoxia-inducible factors in mammals. While vascular endothelial growth factor A (VEGFA) is critical for angiogenesis, VEGFC and its receptor VEGF receptor-3 (VEGFR-3) are essential for the initial sprouting and directed migration as well as for the subsequent survival of LECs. The aim of this study was to determine the effects of hypoxia on the development of the lymphatic system in zebrafish. Zebrafish embryos were obtained from Tg(SAGFF27C; UAS:GFP) animals carrying a lymphatic reporter gene coupled to green fluorescent protein (GFP). Exposure of 1-day old zebrafish embryos to hypoxic conditions (5% O2) for 24 h inhibited thoracic duct formation (−27%, p < 0.0001). Hypoxia inhibited the expression of pro-lymphangiogenic factors prox1a, vegfc and vegfr-3. This inhibition was relieved after re-oxygenation. On the other hand, hypoxia increased the expression of vegfa, a pro-angiogenic factor. In conclusion, hypoxia has opposite effects on vascular development in zebrafish, inhibiting the development of the lymphatic vascular system while promoting the development of the blood vascular system.

PECULIARITIES OF ENDOTHELIAL CD31 MARKER EXPRESSION IN FETAL HUMAN HEART

In this paper by the results of immunohistochemical, morphometric studies of vascular endothelium in the heart of 21 human fetuses 18-24 weeks gestation topological differences in the spatial distribution of vascular elements in the ventricles and atria have been determined. Considering selective specificity of the endothelial marker CD 31, the morphometric analysis of the relative area of blood vessels in the fetus human heart was first carried out. The obtained morphological data greatly extend the idea concerning the development of coronary vascular system in the prenatal period of ontogenesis.

Reproductive biology and embryonal development of pacific capelin &lt;i&gt;Mallotus villosus catervarius &lt;/i&gt;from the Tauiskaya Bay (northern part of the Okhotsk Sea)

Reproductive biology of embryogeny and early ontogenesis is considered for pacific capelin Mallotus villosus catervarius (Pennant) from the Tauiskaya Bay in the northern Okhotsk Sea on the data of long-term sampling. The stages of periblastic sinus and Kupffer’s vesicle are described for the first time for the genus Mallotus. Results of the study allow to assume development of the embryonic vascular system of respiration for the capelin embryo. Tendency to the pacific capelin fecundity increasing is observed in the last five decades conditioned by its abundance lowering and consequent increase of the older fish percentage.

Maternal Vitamin A Supplementation Expands Adipose Progenitor Population through Promoting Vascular Development

Retinoic acid regulates various types of progenitor differentiation and tissue development including adipose development. However, the impact of maternal vitamin A supplementation on fetal adipose development and its long‐term consequence in adulthood remain unclear. To address, we supplemented maternal mice with 0, 15 or 30 IU/mL retinyl palmitate in water during gestation and lactation. Adipose tissue and serum of weanling offspring, and E7.5, 12.5 and 18.5 fetuses were collected. We found that retinoic acid is the main retinoid in fetuses that was elevated by maternal vitamin A supplementation. The populations of platelet‐derived growth factor receptor alpha positive (PDGFRα+) adipocyte progenitors in both fetuses and weaned offspring were increased due to vitamin A supplementation, which were correlated with heightened vascular density in adipose tissue at weaning. We further found that retinoic acid upregulated vascular endothelial growth factor α (VEGFα) which might explain the enhanced vasculogenesis in adipose tissue of fetuses supplemented with vitamin A. Using florescence tracing, we found that, under cold or retinoic acid stimulation, PDGFRα+ progenitor cells were located in neovascular system. In summary, our results reveal that maternal vitamin A supplementation increases offspring adipose progenitor populations by promoting vascular system development during the fetal stage, which is mediated by retinoic acid receptor signaling.Support or Funding InformationSupported by NIH R01HD067449 and USDA‐NIFA 2015‐67015‐23219

5-Hydroxymethylcytosine-mediated alteration of transposon activity associated with the exposure to adverse in utero environments in human.

Preeclampsia and gestational diabetes mellitus (GDM) are the most common clinical conditions in pregnancy that could result in adverse in utero environments. Fetal exposure to poor environments may raise the long-term risk of postnatal disorders, while epigenetic modifications could be involved. Recent research has implicated involvement of 5-hydroxymethylcytosine (5hmC), a DNA base derived from 5-methylcytosine, via oxidation by ten-eleven translocation (TET) enzymes, in DNA methylation-related plasticity. Here, we show that the TET2 expression and 5hmC abundance are significantly altered in the umbilical veins of GDM and preeclampsia. Genome-wide profiling of 5hmC revealed its specific reduction on intragenic regions from both GDM and preeclampsia compared to healthy controls. Gene Ontology analysis using loci bearing unique GDM- and preeclampsia-specific loss-of-5hmC indicated its impact on several critical biological pathways. Interestingly, the substantial alteration of 5hmC on several transposons and repetitive elements led to their differential expression. The alteration of TET expression, 5hmC levels and 5hmC-mediated transposon activity was further confirmed using established hypoxia cell culture model, which could be rescued by vitamin C, a known activator of TET proteins. Together, these results suggest that adverse pregnancy environments could influence 5hmC-mediated epigenetic profile and contribute to abnormal development of fetal vascular systems that may lead to postnatal diseases.

Prenatal hypoxia enhanced angiotensin II-mediated vasoconstriction via increased oxidative signaling in fetal rats.

Toxic factors could cause in utero hypoxia, and prenatal hypoxia (PH) increased incidence of cardiovascular diseases in late life. It is unclear whether/how PH causes vascular injury during fetal life. This study found that PH significantly increased angiotensin II (Ang II)-mediated vessel contractions in fetal thoracic aortas, which was blocked by losartan, not PD123319, indicating that AT1 receptors played a dominant role in the enhanced fetal vasoconstriction following hypoxia. Prenatal hypoxia increased superoxide production and decreased superoxide dismutase (SOD) expression, associated with the enhanced NADPH oxidase (Nox) 4, but not Nox1 or Nox2 in fetal aortas. Ang II-increased vasoconstriction was inhibited by Nox inhibitor apocynin and SOD mimetic blocker tempol. These findings suggested that PH resulted in Ang II/AT1R-mediated fetal vascular hypertensive re-activity via pathways of Nox4-dependent oxidative stress, providing new information regarding the impact of PH on the functional and molecular development of fetal vascular systems.

Congenital coronary artery anomalies: a bridge from embryology to anatomy and pathophysiology—a position statement of the development, anatomy, and pathology ESC Working Group

Congenital coronary artery anomalies are of major significance in clinical cardiology and cardiac surgery due to their association with myocardial ischaemia and sudden death. Such anomalies are detectable by imaging modalities and, according to various definitions, their prevalence ranges from 0.21 to 5.79%. This consensus document from the Development, Anatomy and Pathology Working Group of the European Society of Cardiology aims to provide: (i) a definition of normality that refers to essential anatomical and embryological features of coronary vessels, based on the integrated analysis of studies of normal and abnormal coronary embryogenesis and pathophysiology; (ii) an animal model-based systematic survey of the molecular and cellular mechanisms that regulate coronary blood vessel development; (iii) an organization of the wide spectrum of coronary artery anomalies, according to a comprehensive anatomical and embryological classification scheme; (iv) current knowledge of the pathophysiological mechanisms underlying symptoms and signs of coronary artery anomalies, with diagnostic and therapeutic implications. This document identifies the mosaic-like embryonic development of the coronary vascular system, as coronary cell types differentiate from multiple cell sources through an intricate network of molecular signals and haemodynamic cues, as the necessary framework for understanding the complex spectrum of coronary artery anomalies observed in human patients.

Dual Coronary Artery Fistula from Left Anterior Descending and Right Coronary Artery to Pulmonary Trunk in a Patient with Myocardial Infarction—A Case Report

Coronary artery fistulas (CAF) are precapillary communications between a coronary artery and a cardiac chamber or vessel. CAF have been described as the most common hemodynamically significant congenital coronary anomal. However, it remains a relatively uncommon clinical problem. Coronary fistulas originates slightly more common from the right than from the left coronary artery, but the bilateral fistulas—those that originate from both coronary arteries—accounts for only 5% of total cases. These bilateral fistulas have a unique tendency to terminate in the pulmonary artery. More than half of the bilateral and only 17% of unilateral fistulas, terminates in this manner [1]. CAF are believed to be embryological remnants of sinusoidal connections between the lumens of the primitive tubular heart. This was first described by Maude Abbott in 1908 [2]. These fistulas are usually discovered incidentally upon coronary angiography [3]. Their incidence in the overall population is reported about 0.002% and constitutes 0.13% of congenital cardiac lesion, however, they are found in 0.05% to 0.25% of patients who undergo coronary angiography. The most common site of drainage is the right ventricle seen in 41% of patients. Congenital CAFs usually result from abnormal embryological development of the myocardial vascular system. Acquired CAFs are seen after trauma, endovascular procedures like coronary angiography, endomyocardial biopsy etcor cardiac transplantation. True fistulas of the circulatory system are characterized by an ectatic vascular segment that shows aberrant flow connecting two vascular territories governed by large pressure differences. We report a case of double coronary to pulmonary artery fistula discovered during emergent coronary angiography for acute inferior wall ST-elevation myocardial infarction (STEMI) in a patient with no prior cardiac symptoms.

The chronological and topological structural features of the atria vascular system during prenatal period of ontogenesis

Aim of the work was to establish the chronological and regional special features of the atria vascular system development of human fetuses. For this aim there were used the hearts of human fetuses from archive materials of medical department and city hospitals. Methods : Fixation and formation of mounts for further study were carried out according to recommendations [Yurin N.А., Radostin А.I., 1995]. After fixation there were carried out staining of proteins of smoothly muscular α-SMA actin markers and protein of Prox–1transcription factor, Ki-67cells proliferation marker, CD-34 endothelial marker. Using Image-Pro Plus The Proven Solution Version 3.0.00.00 Windows 95/NT program on the microphotographies of atria histological sections there were separated with marker the vascular structures with further calculation of its absolute area that was then presented in percentage terms relative to the general area of histological section. Positively staining cells were calculated using Image-Pro Plus The Proven Solution Version 3.0.00.00 Windows 95/NT program, the separation of vessels was carried out in manual mode then the relative area of vascular component presented in percentage terms was calculated in automatic mode. We also used morphological analysis of corrosive casts of atria vessels by the method of scanning electron microscopy in fetuses 33–40 weeks old. The photography of samples was done in the mode of secondary electron emission. Results : In fetal period of prenatal period of human ontogenesis the development of vascular bed is inseparably linked with morphogenesis of heart in whole and its parts. The general part of vascular component in human fetuses atria varied within 4,9–9,6 % during the considered period. Arterial and lymphatic links of atria vascular bed were determined separately in spite of topological proximity. The part of lymphatic link of atria vascular system had not reliable differences between the right and left atria till the 16 week of fetal period. Conclusions : An asynchronicity of the atria vascularization processes and topological difference of the vascular component in general and on its separate links established during the study are synchronized regularities of formation of definitive construction of the heart coronal vascular system

Ion channels, guidance molecules, intracellular signaling and transcription factors regulating nervous and vascular system development.

Our sophisticated thoughts and behaviors are based on the miraculous development of our complex nervous network system, in which many different types of proteins and signaling cascades are regulated in a temporally and spatially ordered manner. Here we review our recent attempts to grasp the principles of nervous system development in terms of general cellular phenomena and molecules, such as volume-regulated anion channels, intracellular Ca2+ and cyclic nucleotide signaling, the Npas4 transcription factor and the FLRT family of axon guidance molecules. We also present an example illustrating that the same FLRT family may regulate the development of vascular networks as well. The aim of this review is to open up new vistas for understanding the intricacy of nervous and vascular system development.

Connective-Tissue Growth Factor (CTGF/CCN2) Induces Astrogenesis and Fibronectin Expression of Embryonic Neural Cells In Vitro.

Connective-tissue growth factor (CTGF) is a modular secreted protein implicated in multiple cellular events such as chondrogenesis, skeletogenesis, angiogenesis and wound healing. CTGF contains four different structural modules. This modular organization is characteristic of members of the CCN family. The acronym was derived from the first three members discovered, cysteine-rich 61 (CYR61), CTGF and nephroblastoma overexpressed (NOV). CTGF is implicated as a mediator of important cell processes such as adhesion, migration, proliferation and differentiation. Extensive data have shown that CTGF interacts particularly with the TGFβ, WNT and MAPK signaling pathways. The capacity of CTGF to interact with different growth factors lends it an important role during early and late development, especially in the anterior region of the embryo. ctgf knockout mice have several cranio-facial defects, and the skeletal system is also greatly affected due to an impairment of the vascular-system development during chondrogenesis. This study, for the first time, indicated that CTGF is a potent inductor of gliogenesis during development. Our results showed that in vitro addition of recombinant CTGF protein to an embryonic mouse neural precursor cell culture increased the number of GFAP- and GFAP/Nestin-positive cells. Surprisingly, CTGF also increased the number of Sox2-positive cells. Moreover, this induction seemed not to involve cell proliferation. In addition, exogenous CTGF activated p44/42 but not p38 or JNK MAPK signaling, and increased the expression and deposition of the fibronectin extracellular matrix protein. Finally, CTGF was also able to induce GFAP as well as Nestin expression in a human malignant glioma stem cell line, suggesting a possible role in the differentiation process of gliomas. These results implicate ctgf as a key gene for astrogenesis during development, and suggest that its mechanism may involve activation of p44/42 MAPK signaling. Additionally, CTGF-induced differentiation of glioblastoma stem cells into a less-tumorigenic state could increase the chances of successful intervention, since differentiated cells are more vulnerable to cancer treatments.

Dysregulated COL3A1 and RPL8, RPS16, and RPS23 in Disc Degeneration Revealed by Bioinformatics Methods.

Bioinformatics analysis of published microarray data. This study aimed to reveal the possible genes and pathways related to the pathogenesis of disc degeneration (DD) by analyzing the microarray data. DD is one of the main causes of low back pain, which has become an enormous economic burden for society. Gene expression data of annulus cells and nucleus pulposus cells from patients with DD and controls subjects were downloaded from Gene Expression Omnibus. T test and enrichment analysis were used to identify differentially expressed genes (DEGs) and DEGs-associated functions and pathways in DD, respectively. Protein-protein interaction network and module were constructed to analyze the key nodes associated with this disease. A total of 326 DEGs and 35 DEGs were obtained from the annulus cells and nucleus pulposus cells, respectively. The DEGs of DD in annulus cells were mainly involved in translation, cell adhesion, cell death regulation, and skeletal system development whereas the DEGs in nucleus pulposus cells were mainly related to the biological processes of vascular system development, skeletal system development, and enzyme-linked receptor protein signaling pathway. COL3A1 was the common DEG in both annulus cells and nucleus pulposus cells. The genes encode ribosomal proteins (RPL8, RPS16, and RPS23) in module were enriched in biological processes of translation, translation elongation, and RNA processing. The results revealed the involvement of COL3A1 in skeletal system process and RPL8, RPS16, and RPS23 in the protein synthesis processes in the progression of DD, suggesting their potential use in the diagnosis and therapy of DD. N/A.

The development of coronary vascular system

Aim. Set the terms of occurrence and morphological markers of coronary vessels in the embryonic period of human ontogenesis.Material and methods. To realize the aim of our work the embryos of human heart from 5 th to 8 th week of prenatal development period were investigated in the amount of 60. The obtained specimens were evaluated by immunohistochemical study. For this purpose, the original monoclonal antibodies have been used, such as transcription factor Prox-1, cell proliferation marker Ki-67, an endothelial marker CD-34 and smooth-muscle actin (α-SMA). To identify the reaction the solution of chromogen 3-diaminobenzidine tetrachloride was applied, which is manifested in a rich brown color in the sensitive cells of the cardiac wall.Conclusions: The morphological specialization of vascular links of coronary system in the embryonic period has a natural sequence - acquisition of venous properties at first and parallel differentiation of arterial structures. After arteriovenous determination the next phase begins – lymphatic specialization of venous endothelial cells with the formation of lymphatic links of coronary vascular system

Piezo1 ion channel pore properties are dictated by C-terminal region.

Piezo1 and Piezo2 encode mechanically activated cation channels that function as mechanotransducers involved in vascular system development and touch sensing, respectively. Structural features of Piezos remain unknown. Mouse Piezo1 is bioinformatically predicted to have 30–40 transmembrane (TM) domains. Here, we find that nine of the putative inter-transmembrane regions are accessible from the extracellular side. We use chimeras between mPiezo1 and dPiezo to show that ion-permeation properties are conferred by C-terminal region. We further identify a glutamate residue within a conserved region adjacent to the last two putative TM domains of the protein, that when mutated, affects unitary conductance and ion selectivity, and modulates pore block. We propose that this amino acid is either in the pore or closely associates with the pore. Our results describe important structural motifs of this channel family and lay the groundwork for a mechanistic understanding of how Piezos are mechanically gated and conduct ions.

Chronic hypoxia in pregnancy affected vascular tone of renal interlobar arteries in the offspring.

Hypoxia during pregnancy could affect development of fetuses as well as cardiovascular systems in the offspring. This study was the first to demonstrate the influence and related mechanisms of prenatal hypoxia (PH) on renal interlobar arteries (RIA) in the 5-month-old male rat offspring. Following chronic hypoxia during pregnancy, phenylephrine induced significantly higher pressor responses and greater vasoconstrictions in the offspring. Nitric oxide mediated vessel relaxation was altered in the RIA. Phenylephrine-stimulated free intracellular calcium was significantly higher in the RIA of the PH group. The activity and expression of L-type calcium channel (Cav1.2), not T-type calcium channel (Cav3.2), was up-regulated. The whole-cell currents of calcium channels and the currents of Cav1.2 were increased compared with the control. In addition, the whole-cell K+ currents were decreased in the offspring exposed to prenatal hypoxia. Activity of large-conductance Ca2+-activated K+ channels and the expression of MaxiKα was decreased in the PH group. The results provide new information regarding the influence of prenatal hypoxia on the development of the renal vascular system, and possible underlying cellular and ion channel mechanisms involved.

From nerve to blood vessel: a new role of Olfm2 in smooth muscle differentiation from human embryonic stem cell-derived mesenchymal cells.

From nerve to blood vessel: a new role of Olfm2 in smooth muscle differentiation from human embryonic stem cell-derived mesenchymal cells.

Expression Pattern of Early Growth Response Gene 1 during Olive Flounder (Paralichthys olivaceus) Embryonic Development.

ABSTRACTThe early growth response protein 1 (Egr-1) is a widely reported zinc finger protein and a well known transcription factor encoded by the Egr-1 gene, which plays key roles in many aspects of vertebrate embryogenesis and in adult vertebrates. The Egr-1 expression is important in the formation of the gill vascular system in flounders, which develops during the post-hatching phase and is essential for survival during the juvenile period. However, the complete details of Egr-1 expression during embryo development in olive flounder are not available. We assessed the expression patterns of Egr-1 during the early development of olive flounders by using reverse transcription polymerase chain reaction (RT-PCR) analysis. Microscopic observations showed that gill filament formation corresponded with the Egr-1 expression. Thus, we showed that Egr-1 plays a vital role in angiogenesis in the gill filaments during embryogenesis. Further, Egr-1 expression was found to be strong at 5 days after hatching (DAH), in the development of the gill vascular system, and this strong expression level was maintained throughout all the development stages. Our findings have important implications with respect to the biological role of Egr-1 and evolution of the first respiratory blood vessels in the gills of olive flounder. Further studies are required to elucidate the Egr-1-mediated stress response and to decipher the functional role of Egr-1 in developmental stages.

Expression Pattern of Early Growth Response Gene 1 during Olive Flounder (Paralichthys olivaceus) Embryonic Development.

The early growth response protein 1 (Egr-1) is a widely reported zinc finger protein and a well known transcription factor encoded by the Egr-1 gene, which plays key roles in many aspects of vertebrate embryogenesis and in adult vertebrates. The Egr-1 expression is important in the formation of the gill vascular system in flounders, which develops during the post-hatching phase and is essential for survival during the juvenile period. However, the complete details of Egr-1 expression during embryo development in olive flounder are not available. We assessed the expression patterns of Egr-1 during the early development of olive flounders by using reverse transcription polymerase chain reaction (RT-PCR) analysis. Microscopic observations showed that gill filament formation corresponded with the Egr-1 expression. Thus, we showed that Egr-1 plays a vital role in angiogenesis in the gill filaments during embryogenesis. Further, Egr-1 expression was found to be strong at 5 days after hatching (DAH), in the development of the gill vascular system, and this strong expression level was maintained throughout all the development stages. Our findings have important implications with respect to the biological role of Egr-1 and evolution of the first respiratory blood vessels in the gills of olive flounder. Further studies are required to elucidate the Egr-1-mediated stress response and to decipher the functional role of Egr-1 in developmental stages.