Direct actions of androgen, estrogen and anti-Müllerian hormone on primate secondary follicle development in the absence of FSH in vitro

Summary The present study investigated the effects of c-type natriuretic peptide (CNP) on the development of murine preantral follicles during in vitro growth (IVG). Preantral follicles isolated from ovaries of Kunming mice were cultured in vitro. In the culture system, CNP was supplemented in the experimental groups and omitted in the control groups. In Experiment 1, CNP was only supplemented at the early stage and follicle development was evaluated. In Experiments 2 and 3, CNP was supplemented during the whole period of in vitro culture. In Experiment 2, follicle development and oocyte maturity were evaluated. In Experiment 3, follicle development and embryo cleavage after in vitro fertilization (IVF) were assessed. The results showed that in the control groups in all three experiments, granulosa cells migrated from within the follicle and the follicles could not reach the antral stage. In the experimental groups in all three experiments, no migration of granulosa cells was observed and follicle development was assessed as attaining the antral stage, which was significantly superior to that of the control group (P < 0.0001). Oocyte meiotic arrest was effectively maintained, hence giving good developmental competence. In conclusion, CNP supplementation in the culture system during IVG benefited the development of murine preantral follicles.

Download Full-text

Feasibility of Secondary Follicle Isolation, Culture and Achievement of In-Vitro Oocyte Maturation from Superovulated Ovaries: An Experimental Proof-of-Concept Study Using Mice

Journal of Clinical Medicine ◽

10.3390/jcm10132757 ◽

2021 ◽

Vol 10 (13) ◽

pp. 2757

Author(s):

Xia Hao ◽

Amandine Anastácio ◽

Kenny A. Rodriguez-Wallberg

Keyword(s):

Oocyte Maturation ◽

In Vitro Maturation ◽

Follicular Development ◽

Clinical Situation ◽

Experimental Proof ◽

Secondary Follicle ◽

17Β Estradiol ◽

Follicle Size ◽

And Control

Fertility preservation through ovarian stimulation, aiming at cryopreserving mature oocytes or embryos, is sometimes unsuccessful. This clinical situation deserves novel approaches to overcome infertility following cancer treatment in patients facing highly gonadotoxic treatment. In this controlled experimental study, we investigated the feasibility of in-vitro culturing secondary follicles isolated from superovulated ovaries of mice recently treated with gonadotropins. The follicle yields of superovulated ovaries were 45.9% less than in unstimulated controls. Follicles from superovulated ovaries showed faster growth pace during the initial 7 days of culture and secreted more 17β-estradiol by the end of culture vs controls. Parameters reflecting the outcome of follicular development and oocyte maturation competence in vitro were similar between superovulated and control groups, with a similar follicle size at the end of culture and around 70% survival. Nearly half of cultured follicles met the criteria for in-vitro maturation in both groups and approximately 60% of those achieved a mature MII oocyte, similarly in both groups. Over 60% of obtained MII oocytes displayed normal-looking spindle and chromosome configurations, without significant differences between the groups. Using a validated follicle culture system, we demonstrated the feasibility of secondary follicle isolation, in-vitro culture and oocyte maturation with normal spindle and chromosome configurations obtained from superovulated mice ovaries.

Download Full-text

Granulosal and thecal expression of bone morphogenetic protein- and activin-binding protein mRNA transcripts during bovine follicle development and factors modulating their expression in vitro

Reproduction ◽

10.1530/rep-11-0150 ◽

2011 ◽

Vol 142 (4) ◽

pp. 581-591 ◽

Cited By ~ 18

Author(s):

Claire Glister ◽

Leanne Satchell ◽

Phil G Knight

Keyword(s):

Bone Morphogenetic Proteins ◽

Transforming Growth Factor ◽

Ex Vivo ◽

Mrna Level ◽

Transcript Abundance ◽

Transforming Growth Factor Β ◽

Follicle Development ◽

Mrna Levels ◽

Theca Interna

Evidence supports local roles for transforming growth factor β superfamily members including activins and bone morphogenetic proteins (BMP) in follicle development. Access of these ligands to signalling receptors is likely modulated by extracellular binding proteins (BP). In this study, we comparedex vivoexpression of four BPs (chordin, gremlin, noggin and follistatin) in granulosal (GC) and theca interna (TC) compartments of developing bovine antral follicles (1–18 mm). Effects of FSH and IGF on BMP and BP expression by cultured GC, and effects of LH and BMPs on BP expression by cultured TC were also examined. Follicular expression of all four BP transcripts was higher in GC than TC compartments (P<0.001) a finding confirmed by immunohistochemistry. Follicle category affected (P<0.01) gremlin and follistatin mRNA abundance, with a significant cell-type×follicle category interaction for chordin, follistatin and noggin. Noggin transcript abundance was lower (P<0.05) in GC of large ‘E-active’ than ‘E-inactive’ follicles while follistatin mRNA level was higher (P<0.01). FSH enhanced CYP19, FSHR, INHBA and follistatin by GC without affecting BMP or BMP–BP expression. IGF increased CYP19 and follistatin, reduced BMP4, noggin and gremlin but did not affect chordin orFSHRmRNA levels. LH increased TC androgen secretion but had no effect on BMP or BP expression. BMPs uniformly suppressed TC androgen production whilst increasing chordin, noggin and gremlin mRNA levels up to 20-fold (P<0.01). These findings support the hypothesis that extracellular BP, mostly from GC, contribute to the regulation of intrafollicular BMP/activin signalling. Enhancement of thecal BP expression by BMP implies an autoregulatory feedback role to prevent excessive signalling.

Download Full-text

Effects of IGF-I bioavailability on bovine preantral follicular development in vitro

Reproduction ◽

10.1530/rep-06-0382 ◽

2007 ◽

Vol 133 (6) ◽

pp. 1121-1128 ◽

Cited By ~ 55

Author(s):

Fiona H Thomas ◽

Bruce K Campbell ◽

David G Armstrong ◽

Evelyn E Telfer

Keyword(s):

Follicular Development ◽

Follicle Development ◽

Dependent Manner ◽

Igf Binding Proteins ◽

Preantral Follicles ◽

Follicle Diameter ◽

Igf I ◽

Igf Binding ◽

Development In Vitro

The aim of this study was to determine the effect of regulation of IGF-I bioavailability on preantral follicle development in vitro. Bovine preantral follicles were cultured for 6 days in serum-free medium with increasing doses of Long R3 (LR3) IGF-I (an analog with low affinity for IGF-binding proteins (IGFBPs)), or human recombinant IGF-I (hrIGF-I). Follicle diameter and estradiol production were measured every second day. On day 6, ratios of oocyte/follicle diameter and oocyte morphology were assessed by histological examination, and IGFBP-2 and -3 were detected by immunocytochemistry and in situ hybridization respectively. Both types of IGF-I increased follicle diameter in a dose-dependent manner (P < 0.05) and increased estradiol production over control levels (P < 0.05). However, follicles treated with LR3 IGF-I and the highest concentration of hrIGF-I (1000 ng/ml) had smaller oocyte/follicle ratios, and increased oocyte degeneration, compared with controls or follicles treated with physiological concentrations of hrIGF-I (P < 0.05). IGFBPs were detected in cultured preantral follicles, indicating a requirement for regulation of IGF bioavailability during the early stages of follicular development. Specifically, IGFBP-3 mRNA was found to be expressed in oocytes, and IGFBP-2 immunoreactivity was detected in oocytes and granulosa cells of cultured follicles. In summary, the regulation of IGF-I bioavailability by IGFBPs is necessary for the co-ordination of oocyte and follicle development in vitro.

Download Full-text

Refining insulin concentrations in culture medium containing growth factors BMP15 and GDF9: An in vitro study of the effects on follicle development of goats

Animal Reproduction Science ◽

10.1016/j.anireprosci.2017.08.011 ◽

2017 ◽

Vol 185 ◽

pp. 118-127 ◽

Cited By ~ 3

Author(s):

D.J. Dipaz-Berrocal ◽

N.A.R. Sá ◽

D.D. Guerreiro ◽

J.J.H. Celestino ◽

J. Leiva-Revilla ◽

...

Keyword(s):

Growth Factors ◽

Culture Medium ◽

In Vitro Study ◽

Vitro Study ◽

Follicle Development

Download Full-text

2-Hydroxy-4-Methylselenobutanoic Acid (HMSeBA) Promote Follicle Development By Antioxidant Pathway in Gilts

10.21203/rs.3.rs-850421/v1 ◽

2021 ◽

Author(s):

Yanpeng Dong ◽

Sirun Chen ◽

Yalei Liu ◽

Zimei Li ◽

Xinlin Jia ◽

...

Keyword(s):

Gene Expression ◽

Antioxidant Capacity ◽

Granulosa Cells ◽

Body Weight Gain ◽

Control Diet ◽

Negative Control ◽

Follicle Development ◽

Selenium Content ◽

Total Selenium

Abstract Background Dietary 2-hydroxy-4-methylselenobutanoic acid (HMSeBA) supplementation can exert antioxidant effects in poultry, pigs and weaned pigs. However, it is unknown whether HMSeBA could improve the development of follicle by anti-oxidize effects in gilt. This study was conducted to evaluate the effects of dietary HMSeBA supplementation on the follicle development in gilt. A total of 36 gilts were randomly fed the control diet (CON, negative control), Na2SeO3 diet containing 0.3 mg Se/kg (positive control) or the HMSeBA diet containing 0.3 mg Se/kg from weaning to the 19th day after the second estrus. In another study, the effect of HMSeBA on the cells viability, proliferation, release of 17βestradiol (E2 ) and antioxidant capacity were investigated in the mouse ovarian granulosa cells in vitro. Results Results showed that HMSeBA group increased the average daily body weight gain (ADG) and decreased the ratio of feed: gain during day 120 to 176 in gilts ( P < 0.05). The selenium (HMSeBA and Na 2 SeO 3 ) increased the weight of uterine at the third estrus. There was no effect of HMSeBA on the number of large follicles (diameter >5mm), but HMSeBA decreased the gene expression of growth differentiation factor-9 ( GDF-9 ) and bone morphogenetic protein-15 ( BMP-15 ) in cumulus-oocyte complexes (COCs). HMSeBA group increased the total selenium content in serum ( P < 0.05) and liver ( P < 0.01) and tended to increase the total selenium content in ovary ( P = 0.08). HMSeBA group decreased the malondialdehyde (MDA) concentration in the serum, liver and ovary ( P < 0.05), increased the total antioxidant capacity (T-AOC) in the liver, thioredoxin reductase (TrxR) in the ovary ( P < 0.05) and increased the activity of GPx in the serum, liver and ovary ( P < 0.05). Na 2 SeO 3 supplementation decreased MDA and increased the T-AOC in liver, increased the T-SOD and TrxR in the ovary compared with control. At the transcription level, HMSeBA group increased the glutathione peroxidase 2 ( GPx2 ) and TrxR1 ( P < 0.05) expression in the liver, and increased the GPx1 expression ( P < 0.05) in the ovary of gilts compared with Na2SeO3 treatment. Besides, HMSeBA group increased the expressions of superoxide dismutase 1 ( SOD1 ) and Thioredoxin l ( Trx1 ) in the liver. In vitro experiment, HMSeBA improved granulosa cells’ proliferation and E2 secretion ( P < 0.05). HMSeBA and Na 2 SeO 3 both increased the T-AOC and decreased MDA in granulosa cells in vitro. Meanwhile, HMSeBA increased T-SOD, GPx, glutathione reductase (GR) and TrxR activity in granulosa cells in vitro. In addition, HMSeBA up-regulated SOD2 and GPx1 gene expression in the granulosa cells in vitro.Conclusion These results demonstrate directly, HMSeBA was more conducive to absorption and storage of selenium in the liver and ovary in gilt, and beneficial to exert the effect of HMSeBA on the antioxidant function in the liver and ovary of gilt. Moreover, HMSeBA has stronger antioxidant capacity in granular cells in vitro , which is more conducive to promoting follicle development. Therefore, the new type of organic selenium, HMSeBA, could be potentially useful for the control of reproductive processes in gilt.

Download Full-text

Evaluation of amniotic hydrogel’s impact on the development of murine pre-antral follicles in mouse model

Science and Technology Development Journal - Natural Sciences ◽

10.32508/stdjns.v2i4.807 ◽

2019 ◽

Vol 2 (4) ◽

pp. 32-39

Author(s):

Van Ngoc Le Trinh ◽

Hang Thi Kim Tran ◽

Thu Thuy Anh Vo ◽

Tuyet Thi Vi Le ◽

Ha Le Bao Tran

Keyword(s):

In Vitro Maturation ◽

High Demand ◽

Antral Follicles ◽

Secondary Follicle ◽

The Moment ◽

Main Component ◽

Supporting Frame ◽

Mature Process

Global average infertility rate is about 6– 12%, and in Vietnam at around 7.7%. As a result, there is a high demand for treatment, especially for female infertility. In vitro maturation (IVM) was evaluated and proven to be the most popular and promising at the moment. In long-term cultivation, the follicle was observed to extend, therefore, the usage of a supporting frame is quite necessary to maintain follicle’s natural sphere structure as well as completing the mature process. Amniotic membrane is an avascular membrane, composed of collagen, fibronectin, nidogen, proteoglycan, containing a big number of growth – factors with antimicrobial, anti-inflammatory, low immunogenicity and viscoelasticity properties. Amniotic hydrogel owns structure formed with thin fibers to help preserve the main component as collagen, which can turn to gel form at 37 degree Celsius. With those properties, amniotic hydrogel showed high potential as a scaffold for the follicle. When amniotic hydrogel is used as a scaffold for cultivating of secondary follicle (100 – 130 µm), the size of oocyte and follicle increased after 12 days of culturing, along with the formation of antrum. The results demonstrated the possibility to use amniotic hydrogel as a scaffold for the development of the secondary follicle.

Download Full-text

FSH Promotes Rat Follicle Development Through Inhibition of the Hippo Signalling Pathway

10.21203/rs.3.rs-784763/v1 ◽

2021 ◽

Author(s):

Tairen Chen ◽

Mengjing Wu ◽

Yuting Dong ◽

Bin Kong ◽

Yufang Cai ◽

...

Keyword(s):

Western Blot ◽

In Vitro Culture ◽

Granulosa Cells ◽

Signalling Pathway ◽

Control Group ◽

Follicle Development ◽

Follicle Growth ◽

Expression Levels ◽

Hippo Signalling

Abstract Purpose: Whether FSH promotes follicle growth by inhibiting the Hippo signalling pathway.METHODS: Ovaries were cultured in vitro into a control group (no intervention), an FSH group (0.3 IU/mL FSH), and a VP group (10 µg/mL vetiporfin). HE staining and follicle counts were performed at each stage after 3 hours of in vitro culture. Immunohistochemistry was performed to study the expression levels of LATS2, YAP, PLATS2, and PYAP, and their expression levels in each group were also analysed by Western blot.The number of secondary follicles was significantly increased in the FSH group, the arrangement of granulosa cells was neater, the nuclear fixation was reduced, and the number of atretic follicles was decreased in the VP group. The number of secondary follicles was significantly increased, the number of atretic follicles was reduced, and granulosa cell nuclear consolidation was reduced in the VP+FSH group. Immunohistochemistry showed that LATS2 and YAP expression levels were significantly increased and PLATS2 and PYAP expression levels were relatively decreased in the FSH group, PYAP and PLATS2 expression levels were significantly increased and YAP expression was significantly decreased in the VP group, and YAP and LATS2 expression levels were significantly increased and PYAP and PLATS2 expression levels were significantly decreased in the VP+FSH group. By Western blot, LATS2 and YAP were elevated and PYAP and PLAT2 were decreased in the FSH group, LATS2 and YAP were decreased and PYAP and PLATS were significantly elevated in the VP group, and LATS2 and YAP were elevated and PYAP and PLATS2 were decreased in the VP+FSH group.CONCLUSION: FSH promotes follicle development by inhibiting the Hippo signalling pathway.

Download Full-text