Potential physiological involvement of nesfatin-1 in regulating swine granulosa cell functions

2020 ◽  
Vol 32 (3) ◽  
pp. 274
Author(s):  
R. Ciccimarra ◽  
S. Bussolati ◽  
F. Grasselli ◽  
S. Grolli ◽  
M. Paolucci ◽  
...  
Keyword(s):  
Granulosa Cell ◽  
Granulosa Cells ◽  
Metabolic Regulation ◽  
Developmental Stages ◽  
Quantitative Polymerase Chain Reaction ◽  
Ovarian Follicle ◽  
Redox Status ◽  
Gene Encoding ◽  
Cell Functions ◽  
Vessel Growth

Nesfatin-1 has recently been indicated as a pleiotropic molecule that is primarily involved in the metabolic regulation of reproductive functions acting at hypothalamic level. The aim of this study was to explore the local action of nesfatin-1 in swine ovarian follicles. Nucleobindin 2 (NUCB2) was verified using real-time quantitative polymerase chain reaction in swine granulosa cells from different sized follicles and nesfatin-1 was localised by immunohistochemistry in sections of the whole porcine ovary. The effects of different concentrations of nesfatin-1 on cell growth, steroidogenesis and the redox status of granulosa cells were determined invitro. In addition, the effects of nesfatin-1 were evaluated in an angiogenesis bioassay because vessel growth is essential for ovarian follicle function. Immunohistochemistry revealed intense positivity for nesfatin-1 in swine granulosa cells in follicles at all developmental stages. Expression of the gene encoding the precursor protein NUCB2 was higher in granulosa cells from large rather than from medium and small follicles. Further, nesfatin-1 stimulated cell proliferation and progesterone production and interfered with redox status by modifying nitric oxide production and non-enzyme scavenging activity in granulosa cells from large follicles. Moreover, nesfatin-1 exhibited a stimulatory effect on angiogenesis. This study demonstrates, for the first time, that nesfatin-1 is physiologically present in the swine ovarian follicle, where it may impair granulosa cell functions.

Melatonin potentially acts directly on swine ovary by modulating granulosa cell function and angiogenesis

2017 ◽  
Vol 29 (12) ◽  
pp. 2305 ◽  
Author(s):  
Giuseppina Basini ◽  
Simona Bussolati ◽  
Roberta Ciccimarra ◽  
Francesca Grasselli
Keyword(s):  
Granulosa Cell ◽  
Granulosa Cells ◽  
Cell Function ◽  
Ovarian Follicle ◽  
Hormone Release ◽  
Follicle Growth ◽  
Reproductive Effects ◽  
Vessel Growth ◽  
Gonadotrophin Releasing Hormone ◽  
Local Modulation

Melatonin exerts well-known reproductive effects, mainly acting on hypothalamic gonadotrophin-releasing hormone release. More recent data suggest that melatonin acts directly at the ovarian level, even if, at present, these aspects have been only partly investigated. Swine follicular fluid contains melatonin and its concentration is significantly reduced during follicular growth. Therefore, the present study was undertaken to examine the effects of melatonin, used at physiological concentrations, on cultured swine granulosa cells collected from small (<3 mm) and large (>5 mm) follicles on the main parameters of granulosa cell function such as proliferation and steroidogenesis, namely oestradiol 17β and progesterone (P4) production. Moreover, the effects of melatonin on superoxide anion and nitric oxide (NO) generation by swine granulosa cells were also investigated. Finally, since angiogenesis is crucial for follicle growth, the effects of melatonin on new vessel growth were studied. Collected data indicate that melatonin interferes with cultured granulosa cell proliferation and steroidogenesis, specifically in terms of P4 production and NO output. In addition, the events of physiological follicular angiogenesis were stimulated by melatonin as evidenced by angiogenesis bioassay. Therefore, we suggest that physiological melatonin concentrations could potentially be involved in local modulation of swine ovarian follicle function.

scholarly journals Effects of Orexin B on Swine Granulosa and Endothelial Cells

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1812
Author(s):  
Francesca Grasselli ◽  
Simona Bussolati ◽  
Stefano Grolli ◽  
Rosanna Di Lecce ◽  
Cecilia Dall’Aglio ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Cell Growth ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Superoxide Anion ◽  
Ovarian Follicle ◽  
Reducing Power ◽  
Redox Status ◽  
Fibrin Gel

In addition to the well-known central modulatory role of orexins, we recently demonstrated a peripheral involvement in swine granulosa cells for orexin A and in adipose tissue for orexin B (OXB). The aim of present research was to verify immunolocalization of OXB and its potential role in modulating the main features of swine granulosa cells. In particular, we explored the effects on granulosa cell proliferation (through the incorporation of bromodeoxyuridine), cell metabolic activity (as indirect evaluation by the assessment of ATP), steroidogenic activity (by immunoenzymatic examination) and redox status (evaluating the production of superoxide anion by means of the WST test, production of nitric oxide through the use of the Griess test and the non-enzymatic reducing power by FRAP test). Our data point out that OXB does not modify granulosa cell growth, steroidogenesis and superoxide anion generation. On the contrary, the peptide stimulates (p < 0.05) nitric oxide output and non-enzymatic reducing power. Since new vessel growth is crucial for ovarian follicle development, a further aim of this study was to explore the expression of prepro-orexin and the effects of OXB on swine aortic endothelial cells. We found that the peptide is ineffective in modulating cell growth, while it inhibits redox status parameters. In addition, we demonstrated a stimulatory effect on angiogenesis evaluated in fibrin gel angiogenesis assay. Taken together, OXB appears to be potentially involved in the modulation of redox status in granulosa and endothelial cells and we could argue an involvement of the peptide in the follicular angiogenic events.

scholarly journals Gene Expression Profiling Reveals Cyp26b1 to Be an Activin Regulated Gene Involved in Ovarian Granulosa Cell Proliferation

Endocrinology ◽  
2011 ◽  
Vol 152 (1) ◽  
pp. 303-312 ◽  
Author(s):  
Jingjing L. Kipp ◽  
Ann Golebiowski ◽  
Guadalupe Rodriguez ◽  
Michael Demczuk ◽  
Signe M. Kilen ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Developmental Stages ◽  
Ovarian Follicle ◽  
Temporal Correlation ◽  
Activin A ◽  
Ovarian Follicle Development ◽  
Microarray Study ◽  
Ovarian Granulosa Cell

Abstract Activin, a member of the TGF-β superfamily, is an important modulator of FSH synthesis and secretion and is involved in reproductive dysfunctions and cancers. It also regulates ovarian follicle development. To understand the mechanisms and pathways by which activin regulates follicle function, we performed a microarray study and identified 240 activin regulated genes in mouse granulosa cells. The gene most strongly inhibited by activin was Cyp26b1, which encodes a P450 cytochrome enzyme that degrades retinoic acid (RA). Cyp26b1 has been shown to play an important role in male germ cell meiosis, but its expression is largely lost in the ovary around embryonic d 12.5. This study demonstrated that Cyp26b1 mRNA was expressed in granulosa cells of follicles at all postnatal developmental stages. A striking inverse spatial and temporal correlation between Cyp26b1 and activin-βA mRNA expression was observed. Cyp26b1 expression was also elevated in a transgenic mouse model that has decreased activin expression. The Cyp26 inhibitor R115866 stimulated the proliferation of primary cultured mouse granulosa cells, and a similar effect was observed with RA and activin. A pan-RA receptor inhibitor, AGN194310, abolished the stimulatory effect of either RA or activin on granulosa cell proliferation, indicating an involvement of RA receptor-mediated signaling. Overall, this study provides new insights into the mechanisms of activin action in the ovary. We conclude that Cyp26b1 is expressed in the postnatal mouse ovary, regulated by activin, and involved in the control of granulosa cell proliferation.

scholarly journals MiRNA-143 mediates the proliferative signaling pathway of FSH and regulates estradiol production

2017 ◽  
Vol 234 (1) ◽  
pp. 1-14 ◽  
Author(s):  
Li Zhang ◽  
XiaoXin Zhang ◽  
Xuejing Zhang ◽  
Yu Lu ◽  
Lei Li ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Loss Of Function ◽  
Cellular Processes ◽  
Cell Functions ◽  
Genes Expression ◽  
Underlying Mechanisms

MicroRNAs (MiRNAs) play important regulatory roles in many cellular processes. MiR-143 is highly enriched in the mouse ovary, but its roles and underlying mechanisms are not well understood. In the current study, we show that miR-143 is located in granulosa cells of primary, secondary and antral follicles. To explore the specific functions of miR-143, we transfected miR-143 inhibitor into primary cultured granulosa cells to study the loss of function of miR-143 and the results showed that miR-143 silencing significantly increased estradiol production and steroidogenesis-related gene expression. Moreover, our in vivo and in vitro studies showed that follicular stimulating hormone (FSH) significantly decreased miR-143 expression. This function of miR-143 is accomplished by its binding to the 3’-UTR of KRAS mRNA. Furthermore, our results demonstrated that miR-143 acts as a negative regulating molecule mediating the signaling pathway of FSH and affecting estradiol production by targeting KRAS. MiR-143 also negatively acts in regulating granulosa cells proliferation and cell cycle-related genes expression. These findings indicate that miR-143 plays vital roles in FSH-induced estradiol production and granulosa cell proliferation, providing a novel mechanism that involves miRNA in regulating granulosa cell functions.

scholarly journals Theca: the forgotten cell of the ovarian follicle

Reproduction ◽  
2010 ◽  
Vol 140 (4) ◽  
pp. 489-504 ◽  
Author(s):  
J M Young ◽  
A S McNeilly
Keyword(s):  
Granulosa Cells ◽  
Developmental Stages ◽  
Vascular System ◽  
Ovarian Follicle ◽  
Primary Follicle ◽  
Diverse Range ◽  
Theca Cells ◽  
Structural Support ◽  
Highly Active ◽  
Thecal Cells

Theca cells function in a diverse range of necessary roles during folliculogenesis; to synthesize androgens, provide crosstalk with granulosa cells and oocytes during development, and provide structural support of the growing follicle as it progresses through the developmental stages to produce a mature and fertilizable oocyte. Thecal cells are thought to be recruited from surrounding stromal tissue by factors secreted from an activated primary follicle. The precise origin and identity of these recruiting factors are currently not clear, but it appears that thecal recruitment and/or differentiation involves not just one signal, but a complex and tightly controlled combination of multiple factors. It is clear that thecal cells are fundamental for follicular growth, providing all the androgens required by the developing follicle(s) for conversion into estrogens by the granulosa cells. Their function is enabled through the establishment of a vascular system providing communication with the pituitary axis throughout the reproductive cycle, and delivering essential nutrients to these highly active cells. During development, the majority of follicles undergo atresia, and the theca cells are often the final follicular cell type to die. For those follicles that do ovulate, the theca cells then undergo hormone-dependent differentiation into luteinized thecal cells of the corpus luteum. While the theca is an essential component of follicle development and ovulation, we do not yet fully understand the control of recruitment and function of theca cells, an important consideration since their function appears to be altered in certain causes of infertility.

scholarly journals The Phytoestrogen Quercetin Impairs Steroidogenesis and Angiogenesis in Swine Granulosa Cells In Vitro

2009 ◽  
Vol 2009 ◽  
pp. 1-8 ◽  
Author(s):  
Sujen Eleonora Santini ◽  
Giuseppina Basini ◽  
Simona Bussolati ◽  
Francesca Grasselli
Keyword(s):  
Granulosa Cell ◽  
Granulosa Cells ◽  
Follicular Development ◽  
Redox Status ◽  
Negative Influence ◽  
Animal Nutrition ◽  
Follicle Development ◽  
Vascular Endothelial ◽  
Endothelial Growth Factor

Experimental evidence documents that nutritional phytoestrogens may interact with reproductive functions but the exact mechanism of action is still controversial. Since quercetin is one of the main flavonoids in livestock nutrition, we evaluated its possible effects on cultured swine granulosa cell proliferation, steroidogenesis, and redox status. Moreover, since angiogenesis is essential for follicle development, the effect of the flavonoid on Vascular Endothelial Growth Factor output by granulosa cells was also taken into account. Our data evidence that quercetin does not affect granulosa cell growth while it inhibits progesterone production and modifies estradiol production in a dose-related manner. Additionally, the flavonoid interferes with the angiogenic process by inhibiting VEGF production as well as by altering redox status. Since steroidogenesis and angiogenesis are strictly involved in follicular development, these findings appear particularly relevant, pointing out a possible negative influence of quercetin on ovarian physiology. Therefore, the possible reproductive impact of the flavonoid should be carefully considered in animal nutrition.

scholarly journals Defective Gonadotropin-Dependent Ovarian Folliculogenesis and Granulosa Cell Gene Expression in Inhibin-Deficient Mice

Endocrinology ◽  
2010 ◽  
Vol 151 (10) ◽  
pp. 4994-5006 ◽  
Author(s):  
Ankur K. Nagaraja ◽  
Brooke S. Middlebrook ◽  
Saneal Rajanahally ◽  
Michelle Myers ◽  
Qinglei Li ◽  
...  
Keyword(s):  
Gene Expression ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Ovarian Follicle ◽  
Tumor Development ◽  
Cell Communication ◽  
Granulosa Cell Tumor ◽  
Tumor Formation ◽  
Ovarian Follicle Development ◽  
Female Mice

Inhibin-α knockout (Inha−/−) female mice develop sex cord-stromal ovarian cancer with complete penetrance and previous studies demonstrate that the pituitary gonadotropins (FSH and LH) are influential modifiers of granulosa cell tumor development and progression in inhibin-deficient females. Recent studies have demonstrated that Inha−/− ovarian follicles develop precociously to the early antral stage in prepubertal mice without any increase in serum FSH. These studies suggest that in the absence of inhibins, granulosa cells differentiate abnormally and thus at sexual maturity may undergo an abnormal response to gonadotropin signaling contributing to tumor development. To test this hypothesis, we stimulated immature wild-type and Inha−/− female mice with gonadotropin analogs prior to tumor formation and subsequently examined gonadotropin-induced ovarian follicle development as well as preovulatory and human chorionic gonadotropin-induced gene expression changes in granulosa cells. We find that at 3 wk of age, inhibin-deficient ovaries do not show further antral development or undergo cumulus expansion. In addition, there are widespread alterations in the transcriptome of gonadotropin-treated Inha−/− granulosa cells, with significant changes in genes involved in extracellular matrix and cell-cell communication. These data indicate the gonadotropins initiate an improper program of cell differentiation prior to tumor formation in the absence of inhibins.

scholarly journals Proliferation of Rat Granulosa Cells during the Periovulatory Interval

Endocrinology ◽  
2005 ◽  
Vol 146 (1) ◽  
pp. 414-422 ◽  
Author(s):  
Jennifer D. Cannon ◽  
Mary Cherian-Shaw ◽  
Charles L. Chaffin
Keyword(s):  
Cell Proliferation ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Cyclin E ◽  
Ovarian Follicle ◽  
Terminal Differentiation ◽  
Cell Types ◽  
Cyclin D2 ◽  
Protein Levels ◽  
Serum Gonadotropin

Granulosa cell proliferation during luteinization and terminal differentiation has historically been assumed to decline rapidly after an ovulatory stimulus. In contrast, terminal differentiation in other cell types has recently been associated with a transient increase in proliferation, suggesting that this may occur in the ovarian follicle. The goal of the current study was to test the hypothesis that an ovulatory stimulus to rats results in additional granulosa cell proliferation before cell cycle arrest. Immature rats were given a single injection of pregnant mare serum gonadotropin (PMSG) followed by human chorionic gonadotropin (hCG) to initiate periovulatory events. The proportion of granulosa cells in S phase did not change until 12 h after hCG, although the majority of the post-hCG proliferation was localized to cumulus granulosa cells for up to 10 h after hCG. The expression of cyclin D2 mRNA did not decline until 12 h after hCG, although both cyclin-dependent kinase (Cdk)4 and Cdk6 mRNA increased at 6 h. Protein levels of cyclin D2 and Cdk4 did not change as a result of hCG, whereas cyclin E increased 6 h after hCG. Kinase activity of Cdk2 dropped markedly by 4 h after hCG, but a slight increase in activity was evident 6–8 h after hCG. These data suggest that cumulus granulosa cells continue to proliferate for up to 10 h after an ovulatory stimulus, possibly via cyclin E/Cdk2. It is concluded that proliferation is maintained in granulosa cells in the proximity of the oocyte during luteinization of the rat follicle.

scholarly journals Granulosa cell biomarkers to predict pregnancy in ART: pieces to solve the puzzle

Reproduction ◽  
2017 ◽  
Vol 153 (2) ◽  
pp. R69-R83 ◽  
Author(s):  
Richard J Kordus ◽  
Holly A LaVoie
Keyword(s):  
Gene Expression ◽  
Granulosa Cell ◽  
Live Birth ◽  
Granulosa Cells ◽  
Expression Profiles ◽  
Ovarian Follicle ◽  
Critical Discussion ◽  
Developmental Competence ◽  
Published Data

Cumulus and mural granulosa cells of the ovarian follicle surround and interact with the developing oocyte. These follicular cells reflect the oocyte’s overall health and may indicate subsequent developmental competence of embryos. Biomarkers of granulosa cells associated with individual oocytes could potentially be used in assisted reproduction to indicate which embryos have the best chance of implanting in the uterus and completing gestation. In this review, we have performed a comprehensive assessment of the recent literature for human cumulus and mural granulosa cell mRNA biomarkers as they relate to pregnancy and live birth. A critical discussion of variables affecting granulosa gene expression profiles for in vitro fertilization patients, including patient demographics and ovarian stimulation regimens, is presented. Although studies with microarray data were evaluated, this synopsis focuses on expressed genes that have been validated by quantitative RT-PCR. Furthermore, we summarize the current published data that support or refute identified granulosa expressed genes as potential biomarkers of embryos that give rise to ongoing pregnancy and live birth. Finally, we review studies that offer predictive models for embryo selection for uterine transfer based on biomarkers that show differential gene expression.

Cobalt chloride, a hypoxia-mimicking agent, modulates redox status and functional parameters of cultured swine granulosa cells

2005 ◽  
Vol 17 (7) ◽  
pp. 715 ◽  
Author(s):  
Francesca Grasselli ◽  
Giuseppina Basini ◽  
Simona Bussolati ◽  
Federico Bianco
Keyword(s):  
Granulosa Cells ◽  
Cobalt Chloride ◽  
Ovarian Follicle ◽  
Redox Status ◽  
Early Embryo ◽  
Vascular Endothelial ◽  
Chemical Hypoxia ◽  
Endothelial Growth Factor ◽  
O2 Sensing ◽  
Stimulation Of

Hypoxia occurs physiologically during ovarian follicle growth; this deprivation represents a triggering stimulus for the production of vascular endothelial growth factor (VEGF) by proliferating granulosa cells, which are mostly responsible for the growth of the follicle. Moreover, the steroidogenic activity of these cells ensures a receptive environment for the implantation and development of the early embryo. The present paper reports the adaptive response of swine granulosa cells to cobalt chloride (CoCl2), a chemical hypoxia-mimicking agent. The effects of the treatment were evaluated on cell proliferation, steroidogenesis and VEGF production. In addition, because mithocondrial reactive oxygen species (ROS) are possibly involved in O2 sensing, ROS levels and scavenging enzyme activity were investigated. In the present study, CoCl2 had no effect on progesterone production, although it significantly reduced oestradiol synthesis. The addition of CoCl2 to granulosa cell culture stimulated VEGF production and the generation of hydrogen peroxide. Chemical hypoxia had different effects on scavenger enzyme activities: the activity of superoxide dismutase was enhanced, that of peroxidase reduced and catalase activity was unaffected. The net result is a ‘pro-oxidant’ state, which appears to be possibly involved in the stimulation of VEGF production, thus inducing follicular angiogenesis.

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