Expression of VEGF165 and VEGF165b during ovarian follicular development

Abstract

Objective: To investigate the role of vascular endothelial growth factor (VEGF)165a, VEGF165b, and VEGF receptor (VEGFR) in the development of bovine follicles.

Methods: We cultured follicular cells that were collected from small, medium, and large sized bovine follicles with estrogen and measured the expression of VEGF, VEGFR2 and VEGF165b by Western blot analysis and immunofluorescence.

Results: The expression of VEGF165 increased in all follicle sizes and the expression of VEGF165b was increased in the small and large follicles after culturing in an estrogen containing medium. The expression of VEGFR2 was increased in the medium and large follicles after culturing with estrogen for 96 h. VEGF165 was activated at 100 ng/mL estrogen in the large follicles for 96 h. In addition, VEGFR2 was upregulated in the medium and large follicles after treated with 100 ng/mL estrogen for 96 h.

Conclusions: This evidence suggests that the expression of VEGF165 and VEGFR is associated with estrogen stimulation during the development of bovine follicles and in an autocrine or paracrine manner. This reveals an advantage during oocyte maturation in vitro.

Keywords

Ovarian follicles / Vascular endothelial growth factor / VEGF / VEGFR2 / VEGF165b / Estrogen stimulation

Conflict of interest statement

The authors declare no conflicts of interest.

Acknowledgements

We thank professor David Bates from the University of Nottingham for supporting chemical and research machine.

Funding

This study received no extramural funding.

Authors’ contributions

Khwanthana Grataitong carried out study design, manuscript writing, and critical discussion. Chinnarat Changsangfa and Bongkoch Turathum made study design, experiment execution, data analysis, manuscript writing, and critical discussion. Morakot Sroyraya performed experiment execution and manuscript review. All authors contributed to the article and approved the submitted version.

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References

[1] EE T. The development of methods for isolation and culture of preantral follicles from bovine and porcine ovaries. Theriogenology 1996; 45: 101-110.

[2] Berisha B, Schams D. Ovarian function in ruminants. Domest Anim Endocrinol 2005; 29(2): 305-317.

[3] Soker S, Gollamudi-Payne S, Fidder H, Charmahelli H, Klagsbrun M. Inhibition of vascular endothelial growth factor (VEGF)-induced endothelial cell proliferation by a peptide corresponding to the exon 7-encoded domain of VEGF165. J Biol Chem 1997; 272(50): 31582-31588.

[4] Kaczmarek MM, Schams D, Ziecik AJ. Role of vascular endothelial growth factor in ovarian physiology- An overview. Reprod Biol 2005; 5(2): 111-136.

[5] Neufeld G, Cohen T, Gengrinovitch S, Poltorak Z. Vascular endothelial growth factor (VEGF) and its receptors. Faseb J 1999; 13(1): 9-22.

[6] Karaman S, Leppänen VM, Alitalo K. Vascular endothelial growth factor signalling in development and disease. Development 2018; 145(14). doi: 10.1242/dev.151019.

[7] Kawano Y, Zeineh Hasan K, Fukuda J, Mine S, Miyakawa I. Production of vascular endothelial growth factor and angiogenic factor in human follicular fluid. Mol Cell Endocrinol 2003; 202(1-2): 19-23.

[8] Jakimiuk AJ, Nowicka MA, Zagozda M, Koziol K, Lewandowski P, Issat T. High levels of soluble vascular endothelial growth factor receptor 1/sFlt1 and low levels of vascular endothelial growth factor in follicular fluid on the day of oocyte retrieval correlate with ovarian hyperstimulation syndrom regardless of the stimulation protocol. J Physiol Pharmacol 2017; 68(3): 477-484.

[9] Yang MY, Fortune JE. Vascular endothelial growth factor stimulates the primary to secondary follicle transition in bovine follicles in vitro. Mol Reprod Dev 2007; 74(9): 1095-1104.

[10] Krüssel JS, Behr B, Milki AA, Hirchenhain J, Wen Y, Bielfeld P, et al. Vascular endothelial growth factor (VEGF) mRNA splice variants are differentially expressed in human blastocysts. Mol Hum Reprod 2001; 7(1): 57-63.

[11] Harper SJ, Bates DO. VEGF-A splicing: The key to anti-angiogenic therapeutics? Nat Rev Cancer 2008; 8(11): 880-887.

[12] Bates DO, Cui TG, Doughty JM, Winkler M, Sugiono M, Shields JD, et al. VEGF165b, an inhibitory splice variant of vascular endothelial growth factor, is down-regulated in renal cell carcinoma. Cancer Res 2002; 62(14): 4123-4131.

[13] Araújo VR, Silva GM, Duarte AB, Magalhães DM, Almeida AP, Gonçalves RF, et al. Vascular endothelial growth factor-A(165) [VEGF-A(165)] stimulates the in vitro development and oocyte competence of goat preantral follicles. Cell Tissue Res 2011; 346(2): 273-281.

[14] Saraiva MV, Celestino JJ, Araújo VR, Chaves RN, Almeida AP, Lima-Verde IB, et al. Expression of follicle-stimulating hormone receptor (FSHR) in goat ovarian follicles and the impact of sequential culture medium on in vitro development of caprine preantral follicles. Zygote 2011; 19(3): 205-214.

[15] Zimmermann RC, Xiao E, Bohlen P, Ferin M. Administration of antivascular endothelial growth factor receptor 2 antibody in the early follicular phase delays follicular selection and development in the rhesus monkey. Endocrinology 2002; 143(7): 2496-2502.

[16] Bruno JB, Celestino JJ, Lima-Verde IB, Lima LF, Matos MH, Araújo VR, et al. Expression of vascular endothelial growth factor (VEGF) receptor in goat ovaries and improvement of in vitro caprine preantral follicle survival and growth with VEGF. Reprod Fertil Dev 2009; 21(5): 679-687.

[17] Qiu Y, Seager M, Osman A, Castle-Miller J, Bevan H, Tortonese DJ, et al. Ovarian VEGF(165)b expression regulates follicular development, corpus luteum function and fertility. Reproduction 2012; 143(4): 501-511.

[18] Nichols JA, Perego MC, Schütz LF, Hemple AM, Spicer LJ. Hormonal regulation of vascular endothelial growth factor A (VEGFA) gene expression in granulosa and theca cells of cattle1. J Anim Sci 2019; 97(7): 3034-3045.

[19] Kosaka N, Sudo N, Miyamoto A, Shimizu T. Vascular endothelial growth factor (VEGF) suppresses ovarian granulosa cell apoptosis in vitro. Biochem Biophys Res Commun 2007; 363(3): 733-737.

[20] Doyle LK, Walker CA, Donadeu FX. VEGF modulates the effects of gonadotropins in granulosa cells. Domest Anim Endocrinol 2010; 38(3): 127-137.

[21] McFee RM, Artac RA, McFee RM, Clopton DT, Smith RA, Rozell TG, et al. Inhibition of vascular endothelial growth factor receptor signal transduction blocks follicle progression but does not necessarily disrupt vascular development in perinatal rat ovaries. Biol Reprod 2009; 81(5): 966-977.

[22] Danforth DR, Arbogast LK, Ghosh S, Dickerman A, Rofagha R, Friedman C. Vascular endothelial growth factor stimulates preantral follicle growth in the rat ovary. Biol Reprod 2003; 68(5): 1736-1741.

[23] Ravindranath N, Little-Ihrig L, Phillips HS, Ferrara N, Zeleznik AJ. Vascular endothelial growth factor messenger ribonucleic acid expression in the primate ovary. Endocrinology 1992; 131(1): 254-260.

[24] Shweiki D, Itin A, Neufeld G, Gitay-Goren H, Keshet E. Patterns of expression of vascular endothelial growth factor (VEGF) and VEGF receptors in mice suggest a role in hormonally regulated angiogenesis. J Clin Invest 1993; 91(5): 2235-2243.

[25] Taylor RN, Mueller MD. Anti-angiogenic treatment of endometriosis: Biochemical aspects. Gynecol Obstet Invest 2004; 57(1): 54-56.

[26] Wulff C, Wilson H, Wiegand SJ, Rudge JS, Fraser HM. Prevention of thecal angiogenesis, antral follicular growth, and ovulation in the primate by treatment with vascular endothelial growth factor Trap R1R2. Endocrinology 2002; 143(7): 2797-2807.

[27] Zimmermann RC, Xiao E, Husami N, Sauer MV, Lobo R, Kitajewski J, et al. Short-term administration of antivascular endothelial growth factor antibody in the late follicular phase delays follicular development in the rhesus monkey. J Clin Endocrinol Metab 2001; 86(2): 768-772.

[28] Varey AH, Rennel ES, Qiu Y, Bevan HS, Perrin RM, Raffy S, et al. VEGF 165 b, an antiangiogenic VEGF-A isoform, binds and inhibits bevacizumab treatment in experimental colorectal carcinoma: Balance of pro- and antiangiogenic VEGF-A isoforms has implications for therapy. Br J Cancer 2008; 98(8): 1366-1379.

[29] Konopatskaya O, Churchill AJ, Harper SJ, Bates DO, Gardiner TA. VEGF165b, an endogenous C-terminal splice variant of VEGF, inhibits retinal neovascularization in mice. Mol Vis 2006; 12: 626-632.

[30] Hua J, Spee C, Kase S, Rennel ES, Magnussen AL, Qiu Y, et al. Recombinant human VEGF165b inhibits experimental choroidal neovascularization. Invest Ophthalmol Vis Sci 2010; 51(8): 4282-4288.

[31] Rennel E, Waine E, Guan H, Schüler Y, Leenders W, Woolard J, et al. The endogenous anti-angiogenic VEGF isoform, VEGF165b inhibits human tumour growth in mice. Br J Cancer 2008; 98(7): 1250-1257.

[32] Merdzhanova G, Gout S, Keramidas M, Edmond V, Coll JL, Brambilla C, et al. The transcription factor E2F1 and the SR protein SC35 control the ratio of pro-angiogenic versus antiangiogenic isoforms of vascular endothelial growth factor-A to inhibit neovascularization in vivo. Oncogene 2010; 29(39): 5392-5403.

[33] Pritchard-Jones RO, Dunn DB, Qiu Y, Varey AH, Orlando A, Rigby H, et al. Expression of VEGF (xxx)b, the inhibitory isoforms of VEGF, in malignant melanoma. Br J Cancer 2007; 97(2): 223-230.

[34] Shimizu T, Jayawardana BC, Tetsuka M, Miyamoto A. Differential effect of follicle-stimulating hormone and estradiol on expressions of vascular endothelial growth factor (VEGF) 120, VEGF164 and their receptors in bovine granulosa cells. J Reprod Dev 2007; 53(1): 105-112.

[35] Abedal-Majed MA, Kurz SG, Springman SA, McNeel AK, Freetly HC, Largen V, et al. Vascular endothelial growth factor A isoforms modulate follicle development in peripubertal heifers independent of diet through diverse signal transduction pathways. Biol Reprod 2020; 102(3): 680-692.

[36] Stassi AF, Gasser F, Velázquez MML, Belotti EM, Gareis NC, Rey F, et al. Contribution of the VEGF system to the follicular persistence associated with bovine cystic ovaries. Theriogenology 2019; 138: 52-65.

[37] Shimizu T, Miyamoto A. Progesterone induces the expression of vascular endothelial growth factor (VEGF) 120 and Flk-1, its receptor, in bovine granulosa cells. Anim Reprod Sci 2007; 102(3-4): 228-237.