scholarly journals Simulated Microgravity Induces the Proliferative Inhibition and Morphological Changes in Porcine Granulosa Cells

2021 ◽  
Vol 43 (3) ◽  
pp. 2210-2219
Author(s):  
Truong Xuan Dai ◽  
Hoang Nghia Son ◽  
Ho Nguyen Quynh Chi ◽  
Hoang Nghia Quang Huy ◽  
Nguyen Thai Minh ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Simulated Microgravity ◽  
Physiological Function ◽  
Morphological Changes ◽  
Cell Number ◽  
Reproductive Hormones ◽  
Cyclin Dependent Kinase ◽  
Ovarian Cells ◽  
Porcine Granulosa Cells ◽  
M Phase

Astronauts are always faced with serious health problems during prolonged spaceflights. Previous studies have shown that weightlessness significantly affects the physiological function of female astronauts, including a change in reproductive hormones and ovarian cells, such as granulosa and theca cells. However, the effects of microgravity on these cells have not been well characterized, especially in granulosa cells. This study aimed to investigate the effects of simulated microgravity (SMG) on the proliferation and morphology of porcine granulosa cells (pGCs). pGC proliferation from the SMG group was inhibited, demonstrated by the reduced O.D. value and cell density in the WST-1 assay and cell number counting. SMG-induced pGCs exhibited an increased ratio of cells in the G0/G1 phase and a decreased ratio of cells in the S and G2/M phase. Western blot analysis indicated a down-regulation of cyclin D1, cyclin-dependent kinase 4 (cdk4), and cyclin-dependent kinase 6 (cdk6), leading to the prevention of the G1-S transition and inducing the arrest phase. pGCs under the SMG condition showed an increase in nuclear area. This caused a reduction in nuclear shape value in pGCs under the SMG condition. SMG-induced pGCs exhibited different morphologies, including fibroblast-like shape, rhomboid shape, and pebble-like shape. These results revealed that SMG inhibited proliferation and induced morphological changes in pGCs.

scholarly journals Effects of simulated microgravity on the morphology of mouse embryonic fibroblasts (MEFs)

2020 ◽  
Vol 25 (6) ◽  
pp. 2156-2160
Author(s):  
HOANG NGHIA SON ◽  
◽  
HO NGUYEN QUYNH CHI ◽  
LE NGOC PHUONG THANH ◽  
TRUONG THI HAN ◽  
...  
Keyword(s):  
Simulated Microgravity ◽  
Morphological Changes ◽  
Mouse Embryonic Fibroblast ◽  
Phase Cell ◽  
Control Group ◽  
Mouse Embryonic Fibroblasts ◽  
Embryonic Fibroblasts ◽  
Cell Ratio ◽  
M Phase ◽  
Filament Bundles

This study aimed to assess the effects of simulated microgravity on mouse embryonic fibroblast (MEF) morphology. The results showed that the area of MEFs under simulated microgravity was 7843.39 ± 551.31 µm2 which was lower than the control group (9832.72 ± 453.86 µm2). The nuclear area of MEFs under simulated microgravity (290.76 ± 4.58 µm2) and the control group (296.8 ± 4.58 µm2) did not statistically differ. In addition, the nuclear shape value of the MEFs under simulated microgravity and the control group did not statistically differ (0.86 ± 0.006 vs. 0.87 ± 0.003, respectively). The nuclear intensity of MEFs under simulated microgravity (19361 ± 852) was higher than the control group (16997 ± 285). Moreover, the flow cytometry analysis indicated the reduced G0/G1 phase cell ratio and the increased S phase and G2/M phase cell ratio in MEFs under simulated microgravity. Simulated microgravity also induced a decrease in diameter of actin filament bundles of the MEFs under simulated microgravity (1.61 ± 0.33 µm) compared to the control group (1.79 ± 0.32 µm). These results revealed that simulated microgravity is capable of inducing the morphological changes of mouse embryonic fibroblasts.

Inhibin production by porcine granulosa and luteal cells: development and biological validation of a RIA

1989 ◽  
Vol 120 (4) ◽  
pp. 511-518 ◽  
Author(s):  
U. Michel ◽  
H. Jarry ◽  
M. Metten ◽  
W. Wuttke
Keyword(s):  
Detection Limit ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Inhibin A ◽  
Serum Free ◽  
Luteal Cells ◽  
Biological Validation ◽  
Ovarian Cells ◽  
Progesterone Secretion ◽  
Porcine Granulosa Cells

Abstract. We describe the development and biological validation of a radioimmunoassay for immuno- and bioactive porcine inhibin. A synthetic 1-32 porcine inhibin peptide was used to raise an antiserum and Tyr-1-32 peptide as tracer. As standard we employed porcine follicular fluid calibrated with the 1-32 α-inhibin. Medium obtained from serum-free cultured porcine granulosa cells was chromatographed on Superose S-12 and Mono-Q. Resulting fractions were analysed for inhibin bio- and immunoreactivity. It is shown that granulosa cells produce at least two types of bioactive inhibins, one being also immunoactive in our RIA. We studied secretion of immunoreactive inhibin from porcine ovarian cells under various conditions: Inhibin secretion from mature and immature granulosa cells can be stimulated by FSH, whereas hCG enhances inhibin secretion only from mature granulosa cells. During extended time of culture, the capability of granulosa cells to secrete inhibin is reduced. In contrast, progesterone secretion from these cells increases; this is due to spontaneous functional luteinization. This assumption is supported by the low inhibin secretion of luteal cells in comparison to granulosa cells. Intracellular inhibin content in luteal cells is below detection limit of the RIA, whereas granulosa cells contain readily detectable amounts of this hormone.

Direct effects of bromocriptine on the steroidogenic capability of porcine granulosa cells

1992 ◽  
Vol 126 (4) ◽  
pp. 338-344 ◽  
Author(s):  
Shusaku Kamada ◽  
Toshiro Kubota ◽  
Makoto Taguchi ◽  
Takeshi Aso
Keyword(s):  
Direct Effect ◽  
Granulosa Cells ◽  
Inhibitory Effect ◽  
Cell Number ◽  
Dependent Manner ◽  
Direct Effects ◽  
Concentration Dependent Manner ◽  
Ovarian Granulosa Cells ◽  
Estradiol Secretion ◽  
Porcine Granulosa Cells

The direct effects of bromocriptine on steroidogenesis were examined in cultured porcine granulosa cells. The following observations were made with bromocriptine: (1) It significantly increased the basal or FSH-stimulated secretion of progesterone in cultured porcine granulosa cells at concentrations exceeding 10−7 mol/l; (2) its inhibitory effect on basal estradiol secretion was demonstrated; (3) it did not influence cell number in cultured porcine granulosa cells; (4) it increased the extracellular accumulation of cAMP in a concentration-dependent manner; and (5) it did not induce a change in cytosolic free Ca2+ concentration. These findings suggest that bromocriptine exerts a direct effect on steroidogenesis in ovarian granulosa cells.

scholarly journals FGF21 Promotes Proliferation and Estradiol Synthesis in Porcine Granulosa Cells

2021 ◽  
Author(s):  
Yamei Hu ◽  
Xiaoge Zhou ◽  
Shengjie Shi ◽  
Yankun Li ◽  
Liang Huang ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Viability ◽  
Granulosa Cells ◽  
Ovarian Tissue ◽  
Follicular Development ◽  
Cell Number ◽  
Cell Cycle Protein ◽  
Protein Levels ◽  
Porcine Granulosa Cells ◽  
Estradiol Synthesis

Abstract Background: The proliferation and estradiol synthesis in granulosa cells (GCs) directly promotes follicular development. Previous studies had found that FGF21 regulated the hypothalamic-pituitary-gonad axis in response to the control of fertility. However, the functions and mechanisms of FGF21 in GCs are unclear.Results: Here, we found that the mRNA and protein levels of FGF21 in the ovarian tissue of high-yielding sows (Sus scrofa) was higher than that in low-yielding sows. Moreover, FGF21 was predominantly expressed in porcine GCs. Additionally, ELISA assay showed estradiol was significantly increased when overexpression of FGF21 in porcine GCs. Meanwhile, overexpressed FGF21 up-regulated both the mRNA and protein levels of key estradiol synthesis-related genes in porcine GCs, including StAR, CYP11A1 and CYP19A1. Corresponsingly, knockdown FGF21 inhibited estradiol levels and its synthesis-related genes expression. Besides, overexpression of FGF21 promoted the proliferation of porcine GCs, displayed as increasing the percentage of S-phase cells in cell cycle and EdU positive cells, including cell viability, and upregulated cell cycle genes, including cell cycle protein B (Cyclin B) and protein E (Cyclin E). Corresponsingly, knockdown FGF21 in porcine GCs suppressed the cell cycle and cell viability, as well as EdU positive cell number.Conclusions: These findings highlight that FGF21 is associated with the development of GCs and may be a novel underlying regulator of porcine follicular development.

A technique for superfusion of isolated granulosa cells. Dynamics of cAMP production and steroidogenesis in response to gonadotropins

1988 ◽  
Vol 117 (4) ◽  
pp. 497-506 ◽  
Author(s):  
Carl Johanson ◽  
Viktor Johanson
Keyword(s):  
Granulosa Cells ◽  
Culture Medium ◽  
Cell Damage ◽  
Adenosine Monophosphate ◽  
Camp Production ◽  
Morphological Examination ◽  
Ovarian Cells ◽  
Temporal Relationships ◽  
Polycarbonate Membranes ◽  
Estradiol 17Β

Abstract. A superfusion model for isolated ovarian cells was developed and characterized in detail. Granulosa cells isolated from pre-ovulatory rat ovarian follicles were placed in superfusion (perifusion) chambers with a volume of 125 μl. Culture medium was pumped through the chambers, collected in 20-min fractions of 600 μl and analysed for cAMP and steroids. Viability was confirmed by morphological examination. The use of polycarbonate membranes to retain the cells in the chambers was abandoned since the membranes caused severe cell damage. The temporal relationships between gonadotropic stimuli and the release of cyclic 3':5'-adenosine monophosphate (cAMP) and steroids was investigated. Within 10 min FSH elicited transient increase in the release of cAMP and progesterone but had no effect on testosterone or estradiol-17β release. Amplitude and duration of the response in cAMP and progesterone release were correlated to concentration and length of the FSH pulse when these parameters were varied within the ranges 1–100 μg/l and 30–270 min, respectively. Compared with the cAMP response, the progesterone response peaked up to 30 min later and lasted 1 to 2 h longer but could not be extended to more than approximately 6 h, not even with longer FSH pulses. These results could indicate a development of desensitization.

Effect of cortisol on neurophysin I/oxytocin and peptidyl glycine-α-amidating mono-oxygenase mRNA expression in bovine luteal and granulosa cells

2013 ◽  
Vol 16 (2) ◽  
pp. 231-239
Author(s):  
A. Ziolkowska ◽  
J. Mlynarczuk ◽  
J. Kotwica
Keyword(s):  
Mrna Expression ◽  
Granulosa Cells ◽  
Molecular Mechanisms ◽  
Hormone Secretion ◽  
Oestrous Cycle ◽  
Luteal Cells ◽  
Ru 486 ◽  
Ovarian Cells ◽  
Key Genes ◽  
Ovary Function

Abstract Cortisol stimulates the synthesis and secretion of oxytocin (OT) from bovine granulosa and luteal cells, but the molecular mechanisms of cortisol action remain unknown. In this study, granulosa cells or luteal cells from days 1-5 and 11-15 of the oestrous cycle were incubated for 4 or 8 h with cortisol (1x10-5, 1x10-7 M). After testing cell viability and hormone secretion (OT, progesterone, estradiol), we studied the effect of cortisol on mRNA expression for precursor of OT (NP-I/OT) and peptidyl glycine-α-amidating mono-oxygenase (PGA). The influence of RU 486 (1x10-5 M), a progesterone receptor blocker and inhibitor of the glucocorticosteroid receptor (GR), on the expression for both genes was tested. Cortisol increased the mRNA expression for NP-I/OT and PGA in granulosa cells and stimulated the expression for NP-I/OT mRNA in luteal cells obtained from days 1-5 and days 11-15 of the oestrous cycle. Expression for PGA mRNA was increased only in luteal cells from days 11-15 of the oestrous cycle. In addition, RU 486 blocked the cortisol-stimulated mRNA expression for NP-I/OT and PGA in both types of cells. These data suggest that cortisol affects OT synthesis and secretion in bovine ovarian cells, by acting on the expression of key genes, that may impair ovary function.

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