Expression of Anti-Mullerian Hormone Protein during Early Follicular Development in the Primate Ovary in Vivo Is Influenced by Suppression of Gonadotropin Secretion and Inhibition of Vascular Endothelial Growth Factor

Anti-Mullerian hormone (AMH) plays a role during early follicular development and selection. The aim of this study was to determine the pattern of AMH protein expression in the marmoset ovary and to investigate the effects of inhibition of gonadotropins or vascular endothelial growth factor (VEGF) activity on AMH expression in vivo. GnRH antagonist or VEGF Trap, a soluble decoy receptor, was administered on d 0 or 5 of the follicular phase of the cycle, and ovaries were collected at the end of the follicular phase (d 10). AMH protein was expressed in the marmoset ovary in granulosa cells from the primary stage, with the most abundant staining at the preantral and early antral stages. Inhibition of gonadotropin secretion or VEGF activity between d 0–10 of the cycle decreased AMH expression in early preantral follicles (P < 0.01), and AMH expression was decreased in late preantral follicles in the presence of the VEGF Trap (P < 0.01), compared with controls. There was significantly less AMH expression in early antral follicles with both treatments (P < 0.01), and a decrease in the ratio of oocyte-associated/basement-membrane-associated granulosa cell expression of AMH (P < 0.05). When treatments were administered from d 5–10 of the cycle, both VEGF Trap and GnRH antagonist decreased AMH expression in preantral follicles (P < 0.01) but had no significant effect on early antral follicles. In conclusion, VEGF and gonadotropins are involved in the regulation of expression of AMH in the marmoset. This AMH expression may be a marker of abnormal folliculogenesis in the absence of gonadotropin stimulation or functional angiogenesis.

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Thrombospondin-1 Expression Is Increased during Follicular Atresia in the Primate Ovary

Endocrinology ◽

10.1210/en.2007-0835 ◽

2007 ◽

Vol 149 (1) ◽

pp. 185-192 ◽

Cited By ~ 25

Author(s):

Fiona H. Thomas ◽

Helen Wilson ◽

Audrey Silvestri ◽

Hamish M. Fraser

Keyword(s):

Endothelial Cells ◽

Granulosa Cells ◽

Follicular Phase ◽

Follicular Atresia ◽

Gonadotropin Secretion ◽

Antral Follicles ◽

Cd36 Expression ◽

Mrna And Protein Expression ◽

Vegf Trap

Thrombospondin (TSP)-1 is an antiangiogenic extracellular matrix glycoprotein that modulates several aspects of cellular function. The aim of this study was to determine the pattern of TSP-1 mRNA and protein expression as well as expression of its receptor CD36 in the marmoset ovary and to investigate the effects of inhibition of gonadotropins or VEGF activity on TSP-1 and CD36 expression in vivo. GnRH antagonist or VEGF Trap, a soluble decoy receptor, was administered on d 0 of the follicular phase of the cycle, and ovaries were collected at the end of the follicular phase (d 10). TSP-1 mRNA and protein were present in granulosa cells of preantral and antral follicles, with the highest staining at the late secondary and tertiary stages. Moreover, expression of TSP-1 mRNA and protein was significantly increased in tertiary follicles undergoing atresia. CD36 protein was detected in granulosa cells of preantral and antral follicles as well as in endothelial cells of large vessels. Inhibition of gonadotropin secretion or VEGF activity had no effect on TSP-1 expression; however, expression of CD36 protein was inhibited by the VEGF Trap. In conclusion, TSP-1 may be involved in the cessation of angiogenesis in follicles undergoing atresia; alternatively, TSP-1 may act on granulosa and/or endothelial cells to promote follicular atresia in the ovary. Angiogenesis is likely to involve a balance between pro- and antiangiogenic factors. Our results suggest that loss of VEGF activity does not regulate TSP-1 expression directly but may influence TSP-1 activity via down-regulation of the CD36 receptor.

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A Thrombospondin-Mimetic Peptide, ABT-898, Suppresses Angiogenesis and Promotes Follicular Atresia in Pre- and Early-Antral Follicles in Vivo

Endocrinology ◽

10.1210/en.2010-0283 ◽

2010 ◽

Vol 151 (12) ◽

pp. 5905-5915 ◽

Cited By ~ 33

Author(s):

Samantha A. Garside ◽

Jack Henkin ◽

Keith D. Morris ◽

Suzanne M. Norvell ◽

Fiona H. Thomas ◽

...

Keyword(s):

Granulosa Cells ◽

Polycystic Ovary ◽

Follicular Phase ◽

Vascular Endothelial ◽

Ovulatory Cycle ◽

Antral Follicles ◽

Angiogenesis Assay ◽

Endothelial Growth Factor

Using a novel in vitro angiogenesis assay, we previously showed that thrombospondin (TSP)-1 has antiangiogenic effects on rat follicles and induces apoptosis in granulosa cells in vitro. ABT-898 is an octapeptide mimetic of TSP-1 closely related to ABT-510. Here, we demonstrate the inhibitory effects of ABT-898 on follicular angiogenesis and its proapoptotic effect on granulosa cells. To investigate the potential of this peptide to inhibit follicular angiogenesis in vivo, marmoset monkeys were treated with 2.5 mg/kg ABT-898 twice daily throughout the follicular phase of the cycle. Although treatment did not block emergence of dominant follicles, angiogenesis was reduced in preantral and early-antral follicles. Furthermore, the incidence of atresia at these follicle stages was increased. To investigate whether treatment with ABT-898 would interfere with the timing or duration of the normal ovulatory rise in plasma progesterone, marmosets were treated with a depot formulation containing 25 mg ABT-898 at the start of the follicular phase, with a second injection after 2 wk. Despite active concentrations of peptide being maintained in the circulation, no apparent effects on the ovulatory cycle were observed. Taken together, these results indicate that ABT-898 is capable of having a dual effect by inhibiting follicular angiogenesis and promoting atresia of antral follicles in vivo but does not prevent ovulation or induce luteolysis, as has been observed with direct vascular endothelial growth factor inhibitors. These results suggest that ABT-898 could be a novel therapeutic to inhibit abnormal angiogenesis and induce atresia of accumulated follicles in polycystic ovary syndrome.

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Histological study of capuchin monkey (Cebus apella) ovarian follicles

Acta Amazonica ◽

10.1590/s0044-59672004000300015 ◽

2004 ◽

Vol 34 (3) ◽

pp. 495-501 ◽

Cited By ~ 8

Author(s):

Sheyla Farhayldes Souza Domingues ◽

Luiz Viana Diniz ◽

Sonia Helena Costa Furtado ◽

Otavio Mitio Ohashi ◽

David Rondina ◽

...

Keyword(s):

Qualitative Data ◽

Histological Study ◽

Follicular Development ◽

Cebus Apella ◽

Neotropical Primates ◽

Preantral Follicles ◽

Antral Follicles ◽

The Right

The present study aimed to obtain quanti-qualitative data about the follicular ovarian population in Cebus apella females. Seven ovaries were obtained from 4 C. apella adult females. The ovaries were subjected to light microscopy. The number of preantral and antral follicles for each ovary was estimated using the Fractionator method. The preantral follicles were classified into primordial, transitional, primary and secondary follicles. Antral follicles were those that presented an antral cavity. All counted follicles were classified as normal or degenerated. The diameter of the follicles, oocytes and their nuclei were determined to accompany the follicular development. All results were represented as mean ± SE. The number of preantral follicles was 56,938 ± 21,888 and 49,133 ± 26,896 for the right and left ovaries, respectively. The percentage of normal follicles was 80 ± 4.95%. The follicular diameter ranged from 22 ± 0.5 µm to 61.2 ± 4.0 µm. Regarding the antral follicles, the number of normal and degenerate follicles per ovary were 60.0 ± 19.0 and 3 ± 1.8 follicles, respectively. The antral follicular diameter was 514.4 + 56.6 µm. In conclusion, the information obtained in this study can be used as a parameter for subsequent in vivo or in vitro studies about folliculogenesis in non-human neotropical primates of the C. apella species.

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Placental Growth Factor Is Required for Ovulation, Luteinization, and Angiogenesis in Primate Ovulatory Follicles

Endocrinology ◽

10.1210/en.2017-00739 ◽

2017 ◽

Vol 159 (2) ◽

pp. 710-722 ◽

Cited By ~ 9

Author(s):

Hannah R Bender ◽

Heidi A Trau ◽

Diane M Duffy

Keyword(s):

Growth Factor ◽

Granulosa Cell ◽

Follicular Fluid ◽

Follicular Development ◽

Placental Growth Factor ◽

Vascular Endothelial ◽

Follicle Rupture ◽

Ovulatory Follicles ◽

Endothelial Growth Factor

Abstract Placental growth factor (PGF) is member of the vascular endothelial growth factor (VEGF) family of angiogenesis regulators. VEGFA is an established regulator of ovulation and formation of the corpus luteum. To determine whether PGF also mediates aspects of ovulation and luteinization, macaques received gonadotropins to stimulate multiple follicular development. Ovarian biopsies and whole ovaries were collected before (0 hours) and up to 36 hours after human chorionic gonadotropin (hCG) administration to span the ovulatory interval. PGF and VEGFA were expressed by both granulosa cells and theca cells. In follicular fluid, PGF and VEGFA levels were lowest before hCG. PGF levels remained low until 36 hours after hCG administration, when PGF increased sevenfold to reach peak levels. Follicular fluid VEGFA increased threefold to reach peak levels at 12 hours after hCG, then dropped to intermediate levels. To explore the roles of PGF and VEGFA in ovulation, luteinization, and follicular angiogenesis in vivo, antibodies were injected into the follicular fluid of naturally developed monkey follicles; ovariectomy was performed 48 hours after hCG, with ovulation expected about 40 hours after hCG. Intrafollicular injection of control immunoglobulin G resulted in no retained oocytes, follicle rupture, and structural luteinization, including granulosa cell hypertrophy and capillary formation in the granulosa cell layer. PGF antibody injection resulted in oocyte retention, abnormal rupture, and incomplete luteinization, with limited and disorganized angiogenesis. Injection of a VEGFA antibody resulted in oocyte retention and very limited follicle rupture or structural luteinization. These studies demonstrate that PGF, in addition to VEGFA, is required for ovulation, luteinization, and follicular angiogenesis in primates.

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In vitro and in vivo regulation of follicular formation and activation in cattle

Reproduction Fertility and Development ◽

10.1071/rd10250 ◽

2011 ◽

Vol 23 (1) ◽

pp. 15 ◽

Cited By ~ 27

Author(s):

Joanne E. Fortune ◽

Ming Y. Yang ◽

Wanzirai Muruvi

Keyword(s):

Follicular Development ◽

First Trimester ◽

Vascular Endothelial ◽

Second Trimester ◽

Primordial Follicles ◽

Kit Ligand ◽

Secondary Stage ◽

Endothelial Growth Factor

The establishment of a stockpile of non-growing, primordial follicles and its gradual depletion through activation of primordial follicles are essential processes for female fertility. However, the mechanisms that regulate follicle formation, the activation of primordial follicles to begin growth and the primary-to-secondary follicle transition are poorly understood, especially in domestic animals and primates. The authors’ laboratory is engaged in studying early stages of follicular development in cattle and this review summarises the progress to date. Bovine follicles begin to form in fetal ovaries around the beginning of the second trimester of pregnancy (about Day 90), but the first activated, primary follicles do not appear until after Day 140. Bovine fetal ovaries produce steroids and production is highest during the first trimester. In vitro, oestradiol and progesterone inhibit follicle formation and acquisition by newly formed follicles of the capacity to activate. Meiotic arrest of the oocyte in the diplotene stage of first prophase does not occur until after follicle formation and is correlated with acquisition of the capacity to activate. This may explain the gap between follicle formation and appearance of the first activated follicles. Once capacity to activate has been acquired, it seems likely that activation in vivo is controlled by the balance between stimulators and inhibitors of activation. Insulin and kit ligand stimulate and anti-Müllerian hormone (AMH) inhibits activation in vitro. Few bovine follicles transition from the primary to the secondary stage in vitro, but this transition is increased by medium supplements, testosterone and vascular endothelial growth factor (VEGF).

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

Reproduction Fertility and Development ◽

10.1071/rd18508 ◽

2020 ◽

Vol 32 (3) ◽

pp. 259 ◽

Cited By ~ 2

Author(s):

Xiaomeng Gao ◽

Jinbi Zhang ◽

Zengxiang Pan ◽

Qifa Li ◽

Honglin Liu

Keyword(s):

Vascular Endothelial Growth Factor ◽

Growth Factor ◽

Granulosa Cells ◽

Molecular Level ◽

Follicular Development ◽

Vascular Endothelial ◽

Antral Follicles ◽

Physiological Processes ◽

Endothelial Growth Factor

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 (3–5mm 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.

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Oestrogen and progesterone receptors in the marmoset endometrium: changes during the ovulatory cycle, early pregnancy and after inhibition of vascular endothelial growth factor, GnRH or ovariectomy

Reproduction ◽

10.1530/rep-06-0266 ◽

2007 ◽

Vol 134 (2) ◽

pp. 341-353 ◽

Cited By ~ 7

Author(s):

Audrey Silvestri ◽

Hamish M Fraser

Keyword(s):

Vascular Endothelial Growth Factor ◽

Growth Factor ◽

Gnrh Antagonist ◽

Vascular Endothelial ◽

Ovulatory Cycle ◽

Secretory Phase ◽

Proliferative Phase ◽

Endothelial Growth Factor ◽

Vegf Trap ◽

In Pregnancy

Marmosets are widely used, but detailed studies on localisation of endometrial oestrogen receptors α and β (ERα and ERβ ), and the progesterone receptor (PR) are lacking. These receptors were localised and semi-quantitatively analysed throughout the ovulatory cycle, weeks 2, 3 and 4 of pregnancy and after treatment with GnRH antagonist, vascular endothelial growth factor (VEGF) Trap or ovariectomy. The PR in epithelial cells increased markedly between the mid- and late proliferative phases before declining in the mid-secretory phase and pregnancy. PR in stromal cells was present throughout the cycle and levels were maintained in pregnancy. ERα was present at the mid-proliferative phase and increased in glands at the late proliferative and early secretory phases, before declining at the late secretory phase and week 4 of pregnancy. Stromal ERα showed a similar trend, but decreased earlier, by the mid-secretory phase. ERβ was highly expressed in epithelial cells throughout the cycle and in pregnancy. In stroma, increases in ERβ expression were observed at the late proliferative phase with the staining index decreasing by half as the secretory phase progressed and in pregnancy. GnRH antagonist, VEGF Trap or ovariectomy caused significant reductions in PR and ERβ expression, but not in ERα when compared with the late proliferative phase of the normal cycle. Endothelial cells expressed ERβ , but not ERα or PR. It is concluded that the steroid receptor profile in the marmoset endometrium is generally similar to the human and should provide a useful model for studies on hormonal manipulation of the endometrium.

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Regulation and manipulation of angiogenesis in the primate corpus luteum

Reproduction ◽

10.1530/rep.0.1210355 ◽

2001 ◽

pp. 355-362 ◽

Cited By ~ 34

Author(s):

HM Fraser ◽

SF Lunn

Keyword(s):

Vascular Endothelial Growth Factor ◽

Growth Factor ◽

Corpus Luteum ◽

Follicular Development ◽

Vascular Endothelial ◽

Primate Model ◽

Luteal Function ◽

Short Period ◽

Endothelial Growth Factor

Intense physiological angiogenesis occurs during the early stages of luteal development, providing a model in which the complex processes regulating the angiogenic pathway may be studied. Here, a working hypothesis is presented to explain the diverse changes in the vasculature of the corpus luteum that occur over a short period, based around changes in vascular endothelial growth factor, the angiopoietins and matrix metalloproteinases. An illustration is given of how angiogenesis can be monitored in a primate model and how the role of individual angiogenic factors such as vascular endothelial growth factor may be explored in vivo. Because of the marked effect of inhibition of angiogenesis on luteal function, it is predicted that the normal processes of follicular development, ovulation and luteal function could all be profoundly influenced by the manipulation of angiogenesis.

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Follicular development and the expression of BAX and vascular endothelial growth factor in transplanted ovaries in uni- and bilateral ovariectomized mice: An experimental study

International Journal of Reproductive BioMedicine ◽

10.18502/ijrm.v19i4.9062 ◽

2021 ◽

Author(s):

Maryam Dehghan ◽

Shirin Shahbazi ◽

Mojdeh Salehnia

Keyword(s):

Vascular Endothelial Growth Factor ◽

Growth Factor ◽

Follicular Development ◽

Control Group ◽

Vascular Endothelial ◽

Vegf Expression ◽

Intact Control ◽

Antral Follicles ◽

Ovariectomized Mice ◽

Endothelial Growth Factor

Background: Several conflicting results have been reported on the survival and function of transplanted ovaries. Objective: Evaluation of the follicular development and the expression of vascular endothelial growth factor (VEGF) and Bcl-2-associated X protein (BAX) in ovaries transplanted into uni- and bilaterally ovariectomized mice. Materials and Methods: In this experimental study, 40 female NMRI mice (21-days-old, 12-15 gr) were ovariectomized uni- and bilaterally (n = 20/ group), while the 8-wk-old mice were considered as intact control group (n = 6). 5 weeks after transplantation at the proestrus stage, the morphology of recovered transplanted ovaries and the proportion of follicles were studied at different developmental stages. The apoptosis cell death by pro-apoptotic protein BAX and the expression of VEGF were evaluated using immunohistochemistry. Results: In the bilaterally ovariectomized mice, among the 455 counted normal follicles, a lower rate of primordial and primary follicles and a higher rate of preantral and antral follicles were observed (p = 0.002). However, the percentages of preantral and antral follicles, and the corpus luteum were significantly lower in the intact control group (among the 508 counted normal follicles in this group) compared to other transplanted groups (p = 0.002). The number of BAX-positive cells in all groups was not significantly different. The VEGF expression was prominent in vessels of the corpus luteum, and also in the theca layer of large follicles of studied groups. Conclusion: Early discharge of ovarian reserve was prominent in the bilaterally ovariectomized group but the incidence of apoptotic cells and VEGF expression as angiogenic factor did not differ in both ovariectomized mice. Thus, unilaterally ovariectomy has less side effects on the ovarian reserve compared to bilateral ovariectomy. Key words: Autotransplantation, BAX protein, Vascular endothelial growth factor, Ovariectomy, Mice.

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