Changes of Messenger RNA Expression of Angiogenic Factors and Related Receptors During Follicular Development in Gilts1

Shimizu, Takashi; Jiang, Junfeng; Sasada, Hiroshi; Sato, Eimei

doi:10.1095/biolreprod.102.006734

Cited by 75 publications

(94 citation statements)

References 50 publications

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“…X81380): 5 0 -GAAGTGGT-GAAGTTCATGGA-3 0 (forward) and 5 0 -GCCTTGCAACGC-GAGTCTGT-3 0 (reverse) -b-Actin (accession no. U07786, (Shimizu et al 2002)): 5 0 -ATCGTGCGGGACATCAAGGA-3 0 (forward) and 5 0 -AG-GAGGGAGGGCTGGAAGAG-3 0 (reverse).…”

Section: Pcr Analysesmentioning

confidence: 99%

Vascular supply as a discriminating factor for pig preantral follicle selection

Martelli¹,

Bernabò²,

Berardinelli³

et al. 2009

Reproduction

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This research analyses how somatic and vascular compartments change during preantral follicle growth. To address this aim, theca-granulosa (somatic) proliferation indexes (PIs), proportion of proliferating endothelial cells (PE), vascular area (VA) and vascular endothelial growth factor A (VEGFA) expression were simultaneously recorded on single healthy preantral follicles, classified into six different stages on the basis of the diameter and the granulosa layers. An autonomous blood vessel network starts to appear only in class 3. Vascular remodelling requires VEGFA expression, and VEGFA mRNA and VA significantly increase between class 3 and classes 4 and 5 and, further, in class 6. In addition, a positive correlation exists between these parameters in classes 3-5. Despite variation in angiogenesis results from classes 3 to 5, the statistical analysis reveals that the vascular parameters are positively and strictly correlated with somatic PIs. Conversely, class 6, also characterized by higher values of somatic PIs, displays a stable proportion of PEs (y40%) without showing any correlation among the different parameters analysed. To identify follicular subpopulations within different classes, a multivariate hierarchical cluster analysis was performed. This analysis reveals that the majority of classes 3 and 4 are quiescent follicles or structures that grow very slowly. Class 5 represents a transitory category, where half of the follicles maintain a low activity and the remaining express significantly higher levels of granulosa PI and VA. The follicles with this high activity are probably able to reach class 6 becoming dominant structures where somatic and vascular parameters are constantly on high levels and the VA remains the unique differentiating element.

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Section: Pcr Analysesmentioning

confidence: 99%

Vascular supply as a discriminating factor for pig preantral follicle selection

Martelli¹,

Bernabò²,

Berardinelli³

et al. 2009

Reproduction

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“…Actually, the expression and localization of pig VEGF receptors have been analyzed during the follicular growth phase (Shimizu et al 2002) and in CL (Boonyaprakob et al 2003). No information, on the contrary, is available during the transition from the preovulatory to the periovulatory follicular stage.…”

Section: Follicular Vegf In Pig Vascular Remodelling · a Martelli Andmentioning

confidence: 99%

“…To this end, the present study was performed to describe the VEGF spatio-temporal expression pattern and the endothelial cell system involved in blood vessel remodelling during the periovulatory period in prepubertal gilts. In order to describe angiogenesis at a precise phase of follicle development (preovulatory and periovulatory periods), we have therefore adopted a validated hormonal treatment (Mattioli et al 2001, Shimizu et al 2002, Galeati et al 2003) that induces follicle growth by using 1250 IU equine chorionic gonadotrophin (eCG) and stimulates ovulation with 750 IU human chorionic gonadotrophin (hCG).…”

Section: Introductionmentioning

confidence: 99%

Spatio-temporal analysis of vascular endothelial growth factor expression and blood vessel remodelling in pig ovarian follicles during the periovulatory period

Martelli¹,

Berardinelli²,

Russo³

et al. 2006

Journal of Molecular Endocrinology

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Vascular endothelial growth factor (VEGF) expression pattern and blood vessel remodelling were evaluated during the transition from the preovulatory follicle to the corpus luteum (CL). To this end, prepubertal gilts were treated with equine chorionic gonadotrophin (eCG) to collect preovulatory follicles (60 h after eCG) and with human chorionic gonadotrophin (hCG) to obtain periovulatory follicles 18 h and 36 h later. The VEGF mRNA content was analysed by in situ hybridization, while protein localization in follicular fluid (FF) and in granulosa and theca compartments was evaluated by ELISA, immunohistochemistry or western blot. Blood vessel architecture and vascular area (VA) were investigated using immunohistochemistry for von Willenbrand Factor, a specific endothelial marker. Vascular remodelling was finally tested using Ki-67 immunocytochemistry as a proliferation marker, or -smooth muscle actin ( -SMA) as a specific mural cell marker. eCG-treated follicles showed high VEGF levels and two concentric blood vessel networks composed of proliferating endothelial cells without any association with mural components. hCG injection inhibited VEGF synthesis in the granulosa compartment and, as a consequence, the protein fell within the FF. In parallel, endothelial cell proliferation stopped and the VA decreased. Close to ovulation, VEGF production restarted in both follicular compartments and VEGF mRNA content significantly increased in the theca layer. Changes in follicular VEGF secretion were observed; the protein disappeared from FF and was observed in the extracellular matrix. An active angiogenesis characterized the follicle; endothelial cell proliferation was associated with a recruitment of -SMA-positive mural cells. The data presented in this work showed that, in the phases preceding ovulation, a complete vascular remodelling occurs, characterized by both an evident neovascularization and the appearance of blood vessels presenting smooth musculature which could be involved in CL formation after ovulation.

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“…The fms-like-tyrosine kinase-1 (Flt-1) and fetal liver kinase-1 (Flk-1) bind to VEGF with high affinity. The Flt-1 (or VEGFR-1) and Flk-1 (or VEGFR-2) genes are expressed in the granulosa (Berisha et al 2000) and theca (Shimizu et al 2002) cells of follicles in many species in association with the activation of granulosa cell function and the process of follicle growth in the ovary.…”

Section: Introductionmentioning

confidence: 99%

Hormonal regulation and differential expression of neuropilin (NRP)-1 and NRP-2 genes in bovine granulosa cells

Shimizu¹,

Jayawardana²,

Nishimoto³

et al. 2006

Reproduction

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Although much is known about the biology of vascular endothelial growth factor (VEGF) and its receptors, little is known about the role of the VEGF receptors neuropilin (NRP)-1 and NRP-2 in the process of bovine follicle development. The aim of the present study was to examine the hormonal regulation of NRP-1 and NRP-2 mRNAs by real-time PCR in follicles from the bovine ovary and in cultured granulosa cells. The NRP-1 gene was expressed in the granulosa and theca cells in the post-selection (POF) and pre-selection (PRF) follicles in the bovine ovary. In contrast, the NRP-2 gene was expressed only in the theca cells in the POF and the PRF. The level of NRP-1 mRNA was significantly increased by treatment of the cultured granulosa cells with 10 ng/ml estradiol (E2). In contrast, the addition of progesterone (P4) to the culture medium decreased the expression of the NRP-1 gene. The level of NRP-1 mRNA was increased by 10 ng/ml E2 with or without 1 ng/ml P4, but the level of NRP-1 mRNA was decreased if the P4 level was increased to 10 ng/ml, even when 1 ng/ml E2 was also added. Folliclestimulating hormone did not stimulate the expression of the NRP-1 gene. These results are the first data showing that NRP-1, but not NRP-2, is expressed in the granulosa cells of bovine follicles and that NRP-1 gene expression is regulated by sex steroids. Our findings suggest the involvement of NRP-1 in follicle development in the cow.

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Changes of Messenger RNA Expression of Angiogenic Factors and Related Receptors During Follicular Development in Gilts1

Cited by 75 publications

References 50 publications

Vascular supply as a discriminating factor for pig preantral follicle selection

Vascular supply as a discriminating factor for pig preantral follicle selection

Spatio-temporal analysis of vascular endothelial growth factor expression and blood vessel remodelling in pig ovarian follicles during the periovulatory period

Hormonal regulation and differential expression of neuropilin (NRP)-1 and NRP-2 genes in bovine granulosa cells

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