Overexpression of vascular endothelial growth factor and its receptors in bronchial dypslasia demonstrated by quantitative RT-PCR analysis

Vascular endothelial growth factor (VEGF) is one of the pivotal proangiogenic growth factors that has long contributed to our knowledge of blood vessel and circulatory maintenance as well as angiogenesis in both pathology and pathophysiology. However, the non-canonical functions of VEGF in cardiac morphogenesis have not been well characterized. Here, we examined how VEGF regulates cardiomyocyte cell fate. Using chimeric embryos harboring both wild type and VEGF-null embryonic stem cells, we observed that derivatives of VEGF null cells were preferentially recruited to the atrium of the heart in comparison to the ventricles. To further provide physiologic context of this finding, we used reporter-LacZ staining and RT-PCR and found that endogenous VEGF was indeed expressed at much lower levels in the atrium but highly expressed in the ventricle early in cardiac morphogenesis. These data lead to our hypothesis that cell-autonomous expression of VEGF is a determinant of atrial vs. ventricular cardiomyocyte cell fate. To test this hypothesis, we used a VEGF knock-in mouse model of Sm22Cre x Rosa 26 VEGF. VEGF overexpression in cardiomyocytes (and smooth muscle) at E8.5 resulted in lethality by P1 and thickened atrial and ventricular walls in mutant embryos as characterized by histology (H&E, IF). We further explored the molecular changes underlying this phenotype via microarray and RT-PCR and find disruptions in molecular markers necessary for wall development, specifically: Notch-1, BMP10, Nrg-1. Taken together, our data indicates that aberrant embryonic VEGF signaling disrupts several critical signaling pathways and that overexpression leads to disruption of cardiomyocyte proliferation and cardiac morphogenesis. These findings add to the foundation of better understanding heart development, laying the groundwork for future therapy of congenital and acquired cardiac disease.

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Vascular endothelial growth factor regulates germ cell survival during establishment of spermatogenesis in the bovine testis

Reproduction ◽

10.1530/rep-09-0020 ◽

2009 ◽

Vol 138 (4) ◽

pp. 667-677 ◽

Cited By ~ 47

Author(s):

Kyle C Caires ◽

Jeanene de Avila ◽

Derek J McLean

Keyword(s):

Vascular Endothelial Growth Factor ◽

Growth Factor ◽

Germ Cell ◽

Cell Survival ◽

Germ Cells ◽

Pcr Analysis ◽

Vascular Endothelial ◽

Endothelial Growth Factor ◽

Testis Tissue

Vascular endothelial growth factor-A (VEGFA) is a hypoxia-inducible peptide essential for angiogenesis and targets nonvascular cells in a variety of tissues and cell types. The objective of the current study was to determine the function of VEGF during testis development in bulls. We used an explant tissue culture and treatment approach to test the hypothesis that VEGFA-164 could regulate the biological activity of bovine germ cells. We demonstrate that VEGFA, KDR, and FLT1 proteins are expressed in germ and somatic cells in the bovine testis. Treatment of bovine testis tissue with VEGFA in vitro resulted in significantly more germ cells following 5 days of culture when compared with controls. Quantitative real-time RT-PCR analysis determined that VEGF treatment stimulated an intracellular response that prevents germ cell death in bovine testis tissue explants, as indicated by increased expression of BCL2 relative to BAX and decreased expression of BNIP3 at 3, 6, and 24 h during culture. Blocking VEGF activity in vitro using antisera against KDR and VEGF significantly reduced the number of germ cells in VEGF-treated testis tissue to control levels at 120 h. Testis grafting provided in vivo evidence that bovine testis tissue treated with VEGFA for 5 days in culture contained significantly more differentiating germ cells compared with controls. These findings support the conclusion that VEGF supports germ cell survival and sperm production in bulls.

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Real-time RT-PCR quantitative assays and postmortem degradation profiles of erythropoietin, vascular endothelial growth factor and hypoxia-inducible factor 1 alpha mRNA transcripts in forensic autopsy materials

Legal Medicine ◽

10.1016/j.legalmed.2005.09.001 ◽

2006 ◽

Vol 8 (2) ◽

pp. 132-136 ◽

Cited By ~ 40

Author(s):

Dong Zhao ◽

Bao-Li Zhu ◽

Takaki Ishikawa ◽

Li Quan ◽

Dong-Ri Li ◽

...

Keyword(s):

Vascular Endothelial Growth Factor ◽

Growth Factor ◽

Real Time ◽

Hypoxia Inducible Factor ◽

Forensic Autopsy ◽

Vascular Endothelial ◽

Rt Pcr ◽

Hypoxia Inducible Factor 1 ◽

Mrna Transcripts ◽

Endothelial Growth Factor

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Pulmonary Expression of Vascular Endothelial Growth Factor in Sepsis

Archives of Pathology & Laboratory Medicine ◽

10.5858/2003-127-0331-peoveg ◽

2003 ◽

Vol 127 (3) ◽

pp. 331-335 ◽

Cited By ~ 1

Author(s):

Michael Tsokos ◽

Thomas Pufe ◽

Friedrich Paulsen ◽

Sven Anders ◽

Rolf Mentlein

Keyword(s):

Vascular Endothelial Growth Factor ◽

Growth Factor ◽

Alveolar Macrophages ◽

Sepsis Group ◽

Normal Lung ◽

Vascular Endothelial ◽

Rt Pcr ◽

Vegf Expression ◽

Endothelial Growth Factor

Abstract Context.—Vascular endothelial growth factor (VEGF), an angiogenic and chemotactic peptide, is abundantly expressed in normal lung tissue, especially in alveolar and bronchial epithelium, glandular cells of the bronchi, and activated alveolar macrophages. Objective.—To investigate the role of VEGF in progressively impaired lung function as the major complication and cause of death in septic patients. Materials and Methods.—We evaluated pulmonary VEGF expression in lung autopsy material from septic patients who had been cared for by intensive care medicine using enzyme-linked immunosorbent assay (ELISA), reverse transcriptase–polymerase chain reaction (RT-PCR), and immunohistochemical methods. Results.—Compared with expression in nonseptic control individuals (n = 10), pulmonary VEGF expression as determined by ELISA was significantly (P < .001) decreased in septic patients (n = 8). As monitored by RT-PCR, mRNA for the 2 splice variants, VEGF121 and VEGF165, and for VEGFR-2/KDR were expressed in both groups, the yields being lower in the sepsis group. Samples from septic patients lacked or showed only sparse immunoreaction on bronchial and alveolar epithelium, whereas this reaction was strong in all control samples. However, alveolar macrophages were similarly immunopositive in both groups. Conclusions.—The precise underlying mechanisms for the distinctly different expression of pulmonary VEGF in septic patients and nonseptic control individuals are not clear at present. Particularly the role of VEGF in the development of sepsis-induced lung injury and acute respiratory distress syndrome in mechanically ventilated patients suffering from severe sepsis remains to be clarified.

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Renal vascular endothelial growth factor in neonatal obstructive nephropathy. I. Endogenous VEGF

AJP Renal Physiology ◽

10.1152/ajprenal.00293.2005 ◽

2007 ◽

Vol 292 (1) ◽

pp. F158-F167 ◽

Cited By ~ 12

Author(s):

Laura E. Burt ◽

Michael S. Forbes ◽

Barbara A. Thornhill ◽

Susan C. Kiley ◽

Robert L. Chevalier

Keyword(s):

Vascular Endothelial Growth Factor ◽

Growth Factor ◽

Western Blotting ◽

Neonatal Rat ◽

Obstructive Nephropathy ◽

Sham Operation ◽

Vascular Endothelial ◽

Neonatal Rats ◽

Rt Pcr ◽

Endothelial Growth Factor

Obstructive nephropathy constitutes a major cause of renal impairment in children. Chronic unilateral ureteral obstruction (UUO) impairs maturation of the developing kidney and leads to tubular apoptosis and interstitial inflammation. Vascular endothelial growth factor (VEGF) is involved in recovery from various forms of renal injury. We questioned whether the renal expression of endogenous VEGF and its receptor (VEGFR2/Flk-1) is modified by UUO in early development. Neonatal rats were subjected to partial or complete UUO or sham operation. The distribution of immunoreactive VEGF in each kidney was examined after 7, 14, or 28 days. Adult rats were also subjected to sham operation or complete UUO. Tubular VEGF increased between 14 and 28 days in sham-operated rats and in some partially obstructed neonatal rats but decreased with complete UUO. Parallel changes were found by Western blotting, but not by RT-PCR. Immunoreactive VEGF colocalized with mitochondria in proximal and distal tubules and also appeared in type A intercalated cells, glomerular vascular endothelium, and podocytes. While neonatal microvascular renal VEGFR2 receptor staining was strongly positive regardless of UUO, staining was weak in sham-operated adults but increased following UUO. Parallel changes in VEGFR2 expression were verified by RT-PCR and Western blotting. We conclude that endogenous renal VEGF is developmentally regulated in the neonatal rat and is differentially regulated by partial and complete UUO. Following UUO in the adult, the VEGF receptor is upregulated. Endogenous VEGF may serve an adaptive role in responding to tubular injury caused by UUO and may modulate adaptation by the contralateral kidney.

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Systemic Antibiotic and Nonsteroidal Anti-Inflammatory Drug Treatment Decreases the Level of Endogenous Angiogenic Vascular Endothelial Growth Factor in Inflamed Human Periapical Tissues

Applied Sciences ◽

10.3390/app11114976 ◽

2021 ◽

Vol 11 (11) ◽

pp. 4976

Author(s):

Aleksandra Palatyńska-Ulatowska ◽

Marta Michalska ◽

Anna Drelich ◽

Aleksandra Sałagacka-Kubiak ◽

Ewa Balcerczak ◽

...

Keyword(s):

Gene Expression ◽

Vascular Endothelial Growth Factor ◽

Growth Factor ◽

Control Group ◽

Vascular Endothelial ◽

Rt Pcr ◽

Tissue Samples ◽

Inflammatory Drugs ◽

Anti Inflammatory ◽

Endothelial Growth Factor

Vascular endothelial growth factor (VEGF)-induced angiogenesis contributes to inflammatory bone resorption in humans. Widely documented antagonists to resorption include antibiotics and non-steroidal anti-inflammatory drugs (NSAIDs). The purpose of this study was to investigate the effect of these drugs on proangiogenic VEGF levels in periradicular lesions. Periapical tissue biopsies were obtained from 42 patients with chronic periapical periodontitis. VEGF levels were measured using a commercial ELISA kit in patients divided into groups according to treatment: no drugs (control group, n = 25), NSAIDs (n = 7), antibiotics (n = 5), and NSAIDs and antibiotics (n = 5). Reverse transcriptase (RT) reaction was performed in all the samples under analysis. Presence of VEGFA and VEGFB gene expression was assessed using reverse-transcription-polymerase chain reaction (RT-PCR). ELISA analysis indicated that average VEGF levels in tissue samples of patients treated with NSAIDs (6.097 ± 1.930 ng/mL), antibiotics (5.661 ± 2.395 ng/mL), and NSAIDs and antibiotics (7.142 ± 2.601 ng/mL) were significantly lower than in samples of control patients (10.432 ± 4.257 ng/mL, ANOVA p = 0.008). The RT-PCR did not reveal VEGFA gene expression in any of the 42 samples. VEGFB gene expression was found in 26 of 42 samples (69.1%). The use of NSAIDs or antibiotics in patients with exacerbated chronic periodontitis decreases VEGF levels in periapical tissues. Pharmacotherapy may minimize the effects of VEGF on apical periodontitis progression in that way.

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Quantitative RT-PCR as an alternative to late-stage bioassays for vascular endothelial growth factor

Journal of Pharmaceutical and Biomedical Analysis ◽

10.1016/j.jpba.2008.02.011 ◽

2008 ◽

Vol 47 (3) ◽

pp. 460-468 ◽

Cited By ~ 8

Author(s):

Chris J. Burns ◽

Marta M.C.G. Silva ◽

Elaine Gray ◽

C. Jane Robinson

Keyword(s):

Vascular Endothelial Growth Factor ◽

Growth Factor ◽

Late Stage ◽

Vascular Endothelial ◽

Rt Pcr ◽

Endothelial Growth Factor

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Decreased expression of annexin A2 and loss of its association with vascular endothelial growth factor leads to the deficient trophoblastic invasion in preeclampsia

Journal of Basic and Clinical Physiology and Pharmacology ◽

10.1515/jbcpp-2020-0321 ◽

2021 ◽

Vol 0 (0) ◽

Author(s):

Komal Ruikar ◽

Manjunatha Aithala ◽

Praveenkumar Shetty ◽

Udupi Shastry Dinesh ◽

Anil Bargale ◽

...

Keyword(s):

Vascular Endothelial Growth Factor ◽

Growth Factor ◽

Western Blotting ◽

Annexin A2 ◽

Control Group ◽

Decidual Cell ◽

Vascular Endothelial ◽

Rt Pcr ◽

Endothelial Growth Factor ◽

Normotensive Control

Abstract Objectives Preeclampsia (PE) remains the major cause for maternal and foetal mortality and morbidity. Invasion of endovascular trophoblast and remodelling of spiral artery are crucial actions of normal placental development. Non-fulfilment of these processes plays a leading role in the development of preeclampsia. Vascular endothelial growth factor (VEGF) is produced by extravillous trophoblastic tissue and decidual cell population is a well-known angiogenic growth which plays a fundamental role in placental pathogenesis of PE. Annexin A2 (ANXA2) is a profibrinolytic protein receptor required for plasminolysis, which is an important step in the formation of new blood vessel along with VEGF. Role of ANXA2 is poorly studied in context with human reproductive disease like preeclampsia. The purpose of the present study is to examine the expression and association of VEGF and ANXA2 in the term placentas of pregnancies with and without PE. Methods The study group comprised of placental tissues procured from gestations with PE (n=30) and without (n=20) PE. The expression of VEGF and ANXA2 in the placental villous tissue was evaluated quantitatively by means of IHC, western blotting and reverse transcriptase-polymerase chain reaction (RT-PCR). Results Our IHC, western blotting and RT-PCR analysis illustrated the significant decrease in the expression of VEGF and ANXA2 in PE group compared with the normotensive control group (p<0.005). We observed statistically significant positive correlation among the expression of ANXA2 and VEGF in placentas of normotensive control group (p<0.0001). Conclusions The diminished expression of VEGF and ANXA2 in placenta may be associated with the defective angiogenesis and which may possibly play a vital role in PE pathogenesis.

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Characterization of novel VEGF (vascular endothelial growth factor)-C splicing isoforms from mouse

Biochemical Journal ◽

10.1042/bj20100379 ◽

2010 ◽

Vol 428 (3) ◽

pp. 347-358 ◽

Cited By ~ 7

Author(s):

Zhen-Guo Wang ◽

Tipu S. Puri ◽

Richard J. Quigg

Keyword(s):

Vascular Endothelial Growth Factor ◽

Cell Adhesion ◽

Growth Factor ◽

Glutathione Transferase ◽

Pcr Analysis ◽

Vascular Endothelial ◽

Proximal Tubular Epithelial Cells ◽

Splicing Isoforms ◽

Factor C ◽

Endothelial Growth Factor

VEGF (vascular endothelial growth factor)-C is a major growth factor implicated in various physiological processes, such as angiogenesis and lymphangiogenesis. In the present paper, we report the identification of three short VEGF-C splicing isoforms (VEGF-C62, VEGF-C129 and VEGF-C184) from immortalized mouse kidney PTECs (proximal tubular epithelial cells). Semi-quantitative RT (reverse transcription)–PCR analysis showed these isoforms were universally expressed to varying degrees in different tissues with high expression levels in the kidney. In immortalized PTECs and podocytes, VEGF-C62 can activate phosphorylation of FAK (focal adhesion kinase) and promote cell adhesion to substratum. Cell survival was also increased by VEGF-C62 treatment in the absence of serum. VEGF-C62 can also reduce cell proliferation in PTECs and podocytes. Nucleolin was one of the proteins that associated with VEGF-C62 in pull-down assays using GST (glutathione transferase) fusion proteins as bait, indicating different protein binding requirements for VEGF-C62 compared with VEGF-C. In conclusion, these newly identified VEGF-C isoforms represent a new class of proteins, which are potentially involved in epithelial cell adhesion and proliferation through novel receptor pathways.

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