Register      Login
Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

The distribution and expression of vascular endothelial growth factor A (VEGFA) during follicular development and atresia in the pig

Xiaomeng Gao A , Jinbi Zhang https://orcid.org/0000-0002-9492-3425 A C , Zengxiang Pan A B C , Qifa Li A and Honglin Liu A
+ Author Affiliations
- Author Affiliations

A College of Animal Science and Technology, Nanjing Agriculture University, Nanjing 210095, P. R. China.

B National Experimental Teaching Demonstration Center of Animal Science, Nanjing 210095, P. R. China.

C Corresponding authors. Emails: zhangjinbi@njau.edu.cn; owwa@njau.edu.cn

Reproduction, Fertility and Development 32(3) 259-266 https://doi.org/10.1071/RD18508
Submitted: 10 December 2018  Accepted: 19 June 2019   Published: 24 September 2019

Abstract

The involvement of vascular endothelial growth factor A (VEGFA) in ovarian physiological processes has been widely reported, but the location and role of VEGFA during follicular atresia remain unknown. This study investigated the distribution and expression of VEGFA during porcine follicular development and atresia. Pig ovaries were obtained, individual medium-sized (3–5 mm in diameter) antral follicles were separated and classified into healthy, early atretic or progressively atretic groups. Immunobiology and quantitative techniques were used to investigate the varied follicular distribution of VEGFA at both the morphological and molecular level. The results indicated that VEGFA protein expression peaked in tertiary follicles, mostly distributed in the thecal and inner granulosa layers, during follicular development while VEGFA mRNA was mainly expressed in the inner granulosa layers. Additionally, healthy antral follicles showed a significantly higher expression of VEGFA than atretic follicles in both theca and granulosa cells. Knockdown of VEGFA using siRNA revealed an antiapoptosis effect of VEGFA in cultured pig granulosa cells. Our results increase the knowledge of VEGFA functions in follicles.

Additional keywords: follicular angiogenesis, granulosa cell apoptosis, Sus scrofa.


References

Abbas, M. M., Evans, J. J., Sin, I. L., Gooneratne, A., Hill, A., and Benny, P. S. (2003). Vascular endothelial growth factor and leptin: regulation in human cumulus cells and in follicles. Acta Obstet. Gynecol. Scand. 82, 997–1003.
Vascular endothelial growth factor and leptin: regulation in human cumulus cells and in follicles.Crossref | GoogleScholarGoogle Scholar | 14616272PubMed |

Berisha, B., Schams, D., Kosmann, M., Amselgruber, W., and Einspanier, R. (2000). Expression and localisation of vascular endothelial growth factor and basic fibroblast growth factor during the final growth of bovine ovarian follicles. J. Endocrinol. 167, 371–382.
Expression and localisation of vascular endothelial growth factor and basic fibroblast growth factor during the final growth of bovine ovarian follicles.Crossref | GoogleScholarGoogle Scholar | 11115764PubMed |

Bianco, F., Basini, G., Santini, S., and Grasselli, F. (2005). Angiogenic activity of swine granulosa cells: effects of hypoxia and the role of VEGF. Vet. Res. Commun. 29, 157–159.
Angiogenic activity of swine granulosa cells: effects of hypoxia and the role of VEGF.Crossref | GoogleScholarGoogle Scholar | 16244944PubMed |

Cerrillo, M., Rodriguez, S., Mayoral, M., Pacheco, A., Martínez-Salazar, J., and Velasco, J. A. G. (2008). Differential regulation of VEGF and VE-Cadherin by triggering final oocyte maturation with GnRH agonist versus hCG in IVF cycles. Fertil. Steril. 90, S74.
Differential regulation of VEGF and VE-Cadherin by triggering final oocyte maturation with GnRH agonist versus hCG in IVF cycles.Crossref | GoogleScholarGoogle Scholar |

Doyle, L. K., Walker, C. A., and Donadeu, F. X. (2010). VEGF modulates the effects of gonadotropins in granulosa cells. Domest. Anim. Endocrinol. 38, 127–137.
VEGF modulates the effects of gonadotropins in granulosa cells.Crossref | GoogleScholarGoogle Scholar | 19815366PubMed |

Greenwald, G. S. (1989). Temporal and topographic changes in DNA synthesis after induced follicular atresia. Biol. Reprod. 41, 175–181.
Temporal and topographic changes in DNA synthesis after induced follicular atresia.Crossref | GoogleScholarGoogle Scholar | 2804206PubMed |

Huang, Z., and Wells, D. (2010). The human oocyte and cumulus cells relationship: new insights from the cumulus cell transcriptome. Mol. Hum. Reprod. 16, 715–725.
The human oocyte and cumulus cells relationship: new insights from the cumulus cell transcriptome.Crossref | GoogleScholarGoogle Scholar | 20435609PubMed |

Jiang, J. Y., Macchiarelli, G., Tsang, B. K., and Sato, E. (2003). Capillary angiogenesis and degeneration in bovine ovarian antral follicles. Reproduction 125, 211–223.
Capillary angiogenesis and degeneration in bovine ovarian antral follicles.Crossref | GoogleScholarGoogle Scholar | 12578535PubMed |

Kosaka, N., Sudo, N., Miyamoto, A., and Shimizu, T. (2007). Vascular endothelial growth factor (VEGF) suppresses ovarian granulosa cell apoptosis in vitro. Biochem. Biophys. Res. Commun. 363, 733–737.
Vascular endothelial growth factor (VEGF) suppresses ovarian granulosa cell apoptosis in vitro.Crossref | GoogleScholarGoogle Scholar | 17904528PubMed |

Livak, K. J., and Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25, 402–408.
Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method.Crossref | GoogleScholarGoogle Scholar | 11846609PubMed |

López Albors, O., Olsson, F., Llinares, A. B., Gutiérrez, H., Latorre, R., Candanosa, E., Guillén-Martínez, A., and Izquierdo-Rico, M. J. (2017). Expression of the vascular endothelial growth factor system (VEGF) in the porcine oviduct during the estrous cycle. Theriogenology 93, 46–54.
Expression of the vascular endothelial growth factor system (VEGF) in the porcine oviduct during the estrous cycle.Crossref | GoogleScholarGoogle Scholar | 28257866PubMed |

Macchiarelli, G., Nottola, S. A., Vizza, E., Familiari, G., Kikuta, A., Murakami, T., and Motta, P. M. (1993). Microvasculature of growing and atretic follicles in the rabbit ovary: a SEM study of corrosion casts. Arch. Histol. Cytol. 56, 1–12.
Microvasculature of growing and atretic follicles in the rabbit ovary: a SEM study of corrosion casts.Crossref | GoogleScholarGoogle Scholar | 8499120PubMed |

Manabe, N., Goto, Y., Matsudaminehata, F., Inoue, N., Maeda, A., Sakamaki, K., and Miyano, T. (2004). Regulation mechanism of selective atresia in porcine follicles: regulation of granulosa cell apoptosis during atresia. J. Reprod. Dev. 50, 493–514.
Regulation mechanism of selective atresia in porcine follicles: regulation of granulosa cell apoptosis during atresia.Crossref | GoogleScholarGoogle Scholar | 15514456PubMed |

Marchal, R., Vigneron, C., Perreau, C., Balipapp, A., and Mermillod, P. (2002). Effect of follicular size on meiotic and developmental competence of porcine oocytes. Theriogenology 57, 1523–1532.
Effect of follicular size on meiotic and developmental competence of porcine oocytes.Crossref | GoogleScholarGoogle Scholar | 12054210PubMed |

Mattioli, M., Barboni, B., Turriani, M., Galeati, G., Zannoni, A., Castellani, G., Berardinelli, P., Scapolo, P. A., Mattioli, M., and Barboni, B. (2001). Follicle activation involves vascular endothelial growth factor production and increased blood vessel extension. Biol. Reprod. 65, 1014–1019.
Follicle activation involves vascular endothelial growth factor production and increased blood vessel extension.Crossref | GoogleScholarGoogle Scholar | 11566721PubMed |

McFee, R. M., and Cupp, A. S. (2013). Vascular contributions to early ovarian development: potential roles of VEGFA isoforms. Reprod. Fertil. Dev. 25, 333–342.
Vascular contributions to early ovarian development: potential roles of VEGFA isoforms.Crossref | GoogleScholarGoogle Scholar | 23021322PubMed |

Quintana, R., Kopcow, L., Marconi, G., Sueldo, C., Speranza, G., and Barañao, R. I. (2001). Relationship of ovarian stimulation response with vascular endothelial growth factor and degree of granulosa cell apoptosis. Hum. Reprod. 16, 1814–1818.
Relationship of ovarian stimulation response with vascular endothelial growth factor and degree of granulosa cell apoptosis.Crossref | GoogleScholarGoogle Scholar | 11527881PubMed |

Shimizu, T., Jiang, J. Y., Sasada, H., and Sato, E. (2002). Changes of messenger RNA expression of angiogenic factors and related receptors during follicular development in gilts. Biol. Reprod. 67, 1846–1852.
Changes of messenger RNA expression of angiogenic factors and related receptors during follicular development in gilts.Crossref | GoogleScholarGoogle Scholar | 12444062PubMed |

Stouffer, R. L., Martínezchequer, J. C., Molskness, T. A., Xu, F., and Hazzard, T. M. (2001). Regulation and action of angiogenic factors in the primate ovary. Arch. Med. Res. 32, 567–575.
Regulation and action of angiogenic factors in the primate ovary.Crossref | GoogleScholarGoogle Scholar | 11750732PubMed |

Wang, D., Chen, Y., Yang, Y. J., and Jiang, H. Z. (2012). Expression of VEGF in ovary of ovine through the estrous cycle. Chin. J. Vet. Sci. 32, 151–156.

Yamamoto, S., Konishi, I., Tsuruta, Y., Nanbu, K., Mandai, M., Kuroda, H., Matsushita, K., Hamid, A. A., Yura, Y., and Mori, T. (1997). Expression of vascular endothelial growth factor (VEGF) during folliculogenesis and corpus luteum formation in the human ovary. Gynecol. Endocrinol. 11, 371–381.
Expression of vascular endothelial growth factor (VEGF) during folliculogenesis and corpus luteum formation in the human ovary.Crossref | GoogleScholarGoogle Scholar | 9476086PubMed |

Zhang, J., Lin, F., Pan, Z., Xueshan, M. A., Qifa, L. I., and Liu, H. (2013). Comparative study of methods to determine the follicular atresia extent in pigs. Nanjing Nongye Daxue Xuebao 36, 115–119.

Zhang, J., Liu, Y., Yao, W., Li, Q., Liu, H. L., and Pan, Z. (2018). Initiation of follicular atresia: gene networks during early atresia in pig ovaries. Reproduction 156, 23–33.
Initiation of follicular atresia: gene networks during early atresia in pig ovaries.Crossref | GoogleScholarGoogle Scholar | 29743261PubMed |