VEGF modulates the effects of gonadotropins in granulosa cells

Doyle, L.K.; Walker, Catherine; Donadeu, F.X.

doi:10.1016/j.domaniend.2009.08.008

Cited by 41 publications

(30 citation statements)

References 36 publications

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“…Theca cells were harvested by enzymatic digestion of follicular walls as described (Campbell et al 1998). The two cell types were separately washed in culture media and then cultured in plates pre-coated with fibronectin in a humidified atmosphere with 5% CO 2 at 37 8C under serum-free conditions (McCoy's 5a with 0.02 M HEPES, 3 mM L-glutamine, 0.1% (w/v) BSA, 100 IU/ml penicillin and 0.1 mg/ml streptomycin) with 10 ng/ml bovine insulin, 2.5 mg/ml transferrin and 4 ng/ml sodium selenite, as described (Donadeu & Ascoli 2005, Doyle et al 2010.…”

Section: Follicular Cell Culturesmentioning

confidence: 99%

“…E 2 levels in fluid from follicles R4 mm were measured by RIA as described (Doyle et al 2010) without previous extraction of samples. Follicular fluid concentrations of progesterone were measured by a commercial RIA kit (Siemens Healthcare Diagnostics, Surrey, UK) validated in our laboratory by showing parallelism between serial dilutions of ovine miRNAs and ovarian follicular differentiation follicular fluid and the provided assay standard curves.…”

Section: Experimental Animals and Tissue Collectionmentioning

confidence: 99%

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Identification of miRNAs associated with the follicular–luteal transition in the ruminant ovary

McBride

Carré²,

Sontakke³

et al. 2012

Reproduction

167

152

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Little is known about the involvement of microRNAs (miRNAs) in the follicular-luteal transition. The aim of this study was to identify genome-wide changes in miRNAs associated with follicular differentiation in sheep. miRNA libraries were produced from samples collected at defined stages of the ovine oestrous cycle and representing healthy growing follicles, (diameter, 4.0-5.5 mm), pre-ovulatory follicles (6.0-7.0 mm), early corpora lutea (day 3 post-oestrus) and late corpora lutea (day 9). A total of 189 miRNAs reported in sheep or other species and an additional 23 novel miRNAs were identified by sequencing these libraries. miR-21, miR-125b, let-7a and let-7b were the most abundant miRNAs overall, accounting for 40% of all miRNAs sequenced. Examination of changes in cloning frequencies across development identified nine different miRNAs whose expression decreased in association with the follicular-luteal transition and eight miRNAs whose expression increased during this transition. Expression profiles were confirmed by northern analyses, and experimentally validated targets were identified using miRTarBase. A majority of the 29 targets identified represented genes known to be actively involved in regulating follicular differentiation in vivo. Finally, luteinisation of follicular cells in vitro resulted in changes in miRNA levels that were consistent with those identified in vivo, and these changes were temporally associated with changes in the levels of putative miRNA targets in granulosa cells. In conclusion, this is the first study to characterise genome-wide miRNA profiles during different stages of follicle and luteal development. Our data identify a subset of miRNAs that are potentially important regulators of the follicular-luteal transition.

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Section: Follicular Cell Culturesmentioning

confidence: 99%

Section: Experimental Animals and Tissue Collectionmentioning

confidence: 99%

Identification of miRNAs associated with the follicular–luteal transition in the ruminant ovary

McBride

Carré²,

Sontakke³

et al. 2012

Reproduction

167

152

View full text Add to dashboard Cite

show abstract

“…White adipose tissue, which is the major organ of VEGF secretion, surrounds the human ovaries and uterus and assists with maintaining their functions. VEGF has been detected in the follicular fluid of PCOS females undergoing controlled ovarian hyperstimulation, which modulates the effects of gonadotrophins in GCs (8,10). AMH is exclusively produced by ovarian GCs of the developing preantral and antral follicles (19,20).…”

Section: Discussionmentioning

confidence: 99%

“…VEGF is expressed in the ovarian granulosa, theca and granulosa lutein cells (9) and acts by binding to the tyrosine kinase receptors, VEGFR1 and 2 in the ovary. Marked production and secretion of follicular VEGF occur in response to gonadotrophin stimulation (10). Furthermore, hypersecretion of VEGF is frequently observed in patients with polycystic ovarian syndrome (PCOS) (11,12).…”

Section: Introductionmentioning

confidence: 99%

Vascular endothelial growth factor induces anti-Müllerian hormone receptor 2 overexpression in ovarian granulosa cells of in vitro fertilization/intracytoplasmic sperm injection patients

Yanqiu

Lü

Liu

et al. 2016

Molecular Medicine Reports

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“…VEGF alone does not have an effect on cell proliferation and apoptosis, but this factor intensifies the cytoprotective effect of FSH and estradiol in granulosa cells in culture . VEGF and its receptor VEGFR-2 play important roles in mediating and/or enhancing the effects of gonadotropins on granulosa cells (Doyle et al, 2010). Research shows that RNAi of VEGF using a plasmid-based strategy can stably silence VEGF expression in keloid fibroblasts (Zhang et al, 2008), decrease tumor vascularity and growth in vivo (Shen et al, 2007) and inhibit the growth of murine ovarian cancer cell lines in vitro (Zhang et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

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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VEGF modulates the effects of gonadotropins in granulosa cells

Cited by 41 publications

References 36 publications

Identification of miRNAs associated with the follicular–luteal transition in the ruminant ovary

Identification of miRNAs associated with the follicular–luteal transition in the ruminant ovary

Vascular endothelial growth factor induces anti-Müllerian hormone receptor 2 overexpression in ovarian granulosa cells of in vitro fertilization/intracytoplasmic sperm injection patients

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

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