Vascular endothelial growth factor (VEGF) suppresses ovarian granulosa cell apoptosis in vitro

Kosaka, Naomichi; Sudo, Natsuko; Miyamoto, Akio; Shimizu, Takashi

doi:10.1016/j.bbrc.2007.09.061

Cited by 61 publications

(42 citation statements)

References 23 publications

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“…Bax is said to be responsible for the formation of pores on the mitochondrion by regulating the voltage-dependent anion channel. Our result showing significant inhibition of Bax by VEGFA knockdown was contrary to findings of Kosaka et al (2007), who showed no significant difference. The tumor suppressor protein p53, which has a transcription-independent role in apoptosis, also interacts with Bax to promote its activation as well as insertion into the mitochondrial membrane.…”

Section: Discussioncontrasting

confidence: 99%

“…This method provides a novel tool to study the function of VEGFA and its receptors and apoptosis genes of MGC in vitro. Previous reports have indicated that VEGF and its receptors, VEGFR-1 and VEGFR-2, are expressed in pig follicle cells (Barboni et al, 2000), bovine granulosa cells (Einspanier et al, 2002;Greenaway et al, 2004) and MGC (Wulff et al, 2002) in a coordinated fashion in which they protect these cells from apoptosis, which demonstrates that VEGF functions as a survival factor for ovarian follicle and granulosa cells (Kosaka et al, 2007). Therefore, the inhibition of the VEGF gene by RNAi strategy to study its function in MGC requires an understanding of the related effects of this strategy downstream of the VEGF-related genes.…”

Section: Discussionmentioning

confidence: 95%

“…Unlike VEGF, which is produced by most cell types, research has previously demonstrated that the VEGF receptors Flt-1 and Flk-1 are coexpressed in vascular cells (Ferrara, 2002) including granulosa cells in which they interact to protect these cells against apoptotic cell death, and inhibition of VEGF causes a decrease in Flt and Flk expression (Greenaway et al, 2004;Kosaka et al, 2007). Flt-1 is believed to be expressed in ovarian follicles that consist of oocyte, granulosa cells and the cells of the internal and external theca layer (Okamura et al 2001).…”

Section: Discussionmentioning

confidence: 99%

“…It has been reported that vector-based siRNA can successfully knockdown specific gene expression in mammalian cells, including granulosa cells (Zhang et al, 2003;Kosaka et al, 2007). In this study, we constructed four shRNA expressing plasmids targeting the mouse VEGFA gene (VEGF1051, VEGF1359, VEGF1462, and VEGF1487) with the highest score and an shNC (Table 1) …”

Section: Construction Of Shrna Expression Vector Against Vegfmentioning

confidence: 99%

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Inhibition of vascular endothelial growth factor A expression in mouse granulosa cells by lentivector-mediated RNAi

Ally

Zou²,

Jiang³

et al. 2012

Genet. Mol. Res.

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ABSTRACT. Vascular endothelial growth factor (VEGF) has been found responsible for the induction of proliferation and differentiation in granulosa cells. We constructed four short hairpin RNA (shRNA) expression plasmids targeting the mouse VEGFA gene, and examined their effect on VEGF expression in mouse granulosa cells (MGC) in vitro. Four different shRNA oligonucleotides targeting the coding sequence of mouse VEGFA mRNA and one negative control (shNC) were designed and cloned into a pGPU6/GFP/Neo siRNA expression vector, and transiently transfected into MGC. At 48 h post-transfection, total RNA was extracted from the cells and subjected to qRT-PCR analysis. The most effective interference vector, shVEGF1487 was chosen for lentiviral construction. The recombinant plasmid was then transfected into 293FT cells via Lipofectamine TM 2000-mediated gene transfer, for the production of lentivirus, and then concentrated via ultracentrifugation. This lentiviral vector was then used for the transduction of MGC. VEGFA gene expression, apoptosis genes and VEGFA receptor genes were detected by qRT-PCR, the VEGFA protein level in culture media by ELISA assay and protein levels in MGC by Western blot analysis. The four VEGFA expression plasmids were successfully constructed and the most effective interference vector, shVEGF1487, was chosen for lentiviral production and MGC transduction. There was significant knockdown of the VEGFA gene, receptor genes and apoptosis genes for all the shVEGF constructs, compared with the shNC and Mock controls. The lentiviral vector also gave significant knockdown of the VEGFA gene. Protein levels were lower for most of the shVEGFs based on Western blot analysis with exception of VEGF1359; in this case, it was higher than shNC but lower than for the Mock group. Lentivector-transduced MGC also gave lower levels of protein. We conclude that shVEGF expression plasmids and lentivector carrying RNAi are promising tools for the inhibition of VEGF, the corresponding receptor genes, and apoptosis gene expression in MGC.

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Section: Discussioncontrasting

confidence: 99%

Section: Discussionmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 99%

Section: Construction Of Shrna Expression Vector Against Vegfmentioning

confidence: 99%

See 2 more Smart Citations

Inhibition of vascular endothelial growth factor A expression in mouse granulosa cells by lentivector-mediated RNAi

Ally

Zou²,

Jiang³

et al. 2012

Genet. Mol. Res.

View full text Add to dashboard Cite

show abstract

“…VEGF is a key regulator of physiological angiogenesis during embryogenesis, skeletal growth and reproductive functions. Evidences demonstrate that VEGF and its receptors protect follicle and granulosa cells from apoptosis, suggesting that VEGF functions as a survival factor (Kosaka et al, 2007). In this study we identified under expression of members of VEGF pathway, including PI3K and Fig.…”

Section: Discussionmentioning

confidence: 92%

Deregulation of key signaling pathways involved in oocyte maturation in FMR1 premutation carriers with Fragile X-associated primary ovarian insufficiency

Álvarez-Mora

Rodríguez‐Revenga

Madrigal

et al. 2015

Gene

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Co‐ordinated expression of the VEGF system components in granulosa cells to develop a proangiogenic autocrine milieu during ovarian follicle development

Zamora‐Gutiérrez

Guzmán

Hernández‐Coronado

et al. 2018

Molecular Reproduction Devel

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In the present study, we investigated the temporal relationship between angiogenic and antiangiogenic vascular endothelial growth factor isoforms (VEGFxxxa and VEGFxxxb, respectively), the receptors VEGFR1 and VEGFR2, their soluble forms, and the kinases and the splicing factors regulating the synthesis of VEGF isoforms in healthy and atretic antral follicles. The results show a higher (p < 0.05) messenger RNA (mRNA) expression of VEGF120a, VEGF164a, and VEGF120b in healthy than in atretic follicles, but the mRNA expression of VEGF164b was not detected. The mRNA of serine–arginine protein kinase 1 ( SRPK1) was higher ( p < 0.05) in large healthy follicles than in large atretic follicles. In contrast, atretic follicles had higher mRNA expression of a soluble form of the receptor 2 of VEGF ( sVEGFR2) than healthy follicles ( p < 0.05). Additionally, we observed a positive relationship ( p < 0.05) between SRPK1 and serine–arginine‐rich splicing factor 1 ( SRSF1) with the angiogenic isoforms VEGF120a and VEGF164a and between CDC‐like kinases‐1 ( CLK1) and SRSF6 with the antiangiogenic VEGF120b isoform. Principal components analysis (PCA) resulted in two PC explaining 71% of the variation, which was formed by the VEGF isoforms, the kinases and the splicing factor (PC1) and by the VEGF receptors (PC2). When PC analysis was carried out within follicular health status, there were no differences for PC1 between follicular status, whereas PC2 differed between healthy and atretic follicles. In conclusion, the higher mRNA expression for VEGF120a and VEGF164a, the low expression of sVEGFR2, and absent expression of mRNA for VEGF164b provide evidence of a proangiogenic autocrine milieu to support granulosa cells during follicle development.

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Vascular endothelial growth factor (VEGF) suppresses ovarian granulosa cell apoptosis in vitro

Cited by 61 publications

References 23 publications

Inhibition of vascular endothelial growth factor A expression in mouse granulosa cells by lentivector-mediated RNAi

Inhibition of vascular endothelial growth factor A expression in mouse granulosa cells by lentivector-mediated RNAi

Deregulation of key signaling pathways involved in oocyte maturation in FMR1 premutation carriers with Fragile X-associated primary ovarian insufficiency

Co‐ordinated expression of the VEGF system components in granulosa cells to develop a proangiogenic autocrine milieu during ovarian follicle development

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