scholarly journals THE ROLE OF CUMULUS CELLS AND SERUM IN MOUSE OOCYTE MATURATION IN VITRO

Reproduction ◽  
1973 ◽  
Vol 34 (2) ◽  
pp. 241-245 ◽  
Author(s):  
P. C. CROSS
Keyword(s):  
Oocyte Maturation ◽  
Cumulus Cells ◽  
Mouse Oocyte ◽  
Maturation In Vitro

scholarly journals Hypoxia-inducible factor (HIF1alpha) inhibition modulates cumulus cell function and affects bovine oocyte maturation in vitro†

2020 ◽  
Author(s):  
Aslihan Turhan ◽  
Miguel Tavares Pereira ◽  
Gerhard Schuler ◽  
Ulrich Bleul ◽  
Mariusz P Kowalewski
Keyword(s):  
Oocyte Maturation ◽  
Cell Function ◽  
Hypoxia Inducible Factor ◽  
Cumulus Cell ◽  
Cumulus Cells ◽  
Negative Effects ◽  
Protein Levels ◽  
Maturation In Vitro

Abstract Various metabolic and hormonal factors expressed in cumulus cells are positively correlated with the in vitro maturation (IVM) of oocytes. However, the role of hypoxia sensing both during maturation of cumulus–oocyte complexes (COCs) as well as during the resumption of meiosis remains uncertain. HIF1alpha plays major roles in cellular responses to hypoxia, and here we investigated its role during bovine COC maturation by assessing the expression of related genes in cumulus cells. COCs were divided into the following groups: immature (control), in vitro matured (IVM/control), or matured in the presence of a blocker of HIF1alpha activity (echinomycin, IVM/E). We found an inhibition of cumulus cell expansion in IVM/E, compared with the IVM/control. Transcript levels of several factors (n = 13) were assessed in cumulus cells. Decreased expression of HAS2, TNFAIP6, TMSB4, TMSB10, GATM, GLUT1, CX43, COX2, PTGES, and STAR was found in IVM/E (P < 0.05). Additionally, decreased protein levels were detected for STAR, HAS2, and PCNA (P < 0.05), while activated-Caspase 3 remained unaffected in IVM/E. Progesterone output decreased in IVM/E. The application of PX-478, another blocker of HIF1alpha expression, yielded identical results. Negative effects of HIF1alpha suppression were further observed in the significantly decreased oocyte maturation and blastocyst rates from COCs matured with echinomycin (P < 0.05) or PX-478 (P < 0.05). These results support the importance of HIF1alpha for COC maturation and subsequent embryo development. HIF1alpha is a multidirectional factor controlling intercellular communication within COCs, steroidogenic activity, and oocyte development rates, and exerting effects on blastocyst rates.

scholarly journals Perfluorononanoic acid impedes mouse oocyte maturation by inducing mitochondrial dysfunction and oxidative stress

2021 ◽  
Author(s):  
Xiaofei Jiao ◽  
Ning Liu ◽  
Yiding Xu ◽  
Huanyu Qiao
Keyword(s):  
Oxidative Stress ◽  
Dna Damage ◽  
Oocyte Maturation ◽  
Early Stage ◽  
Spindle Assembly ◽  
Mouse Oocyte ◽  
Toxic Effects ◽  
Germinal Vesicle Breakdown ◽  
Maturation In Vitro

Perfluorononanoic acid (PFNA), a member of PFAS, is frequently detected in human blood and tissues, even in follicular fluid of women. The exposure of PFNA, but not PFOA and PFOS, is positively correlated with miscarriage and increased time to pregnancy. Toxicological studies indicated that PFNA exposure is associated with immunotoxicity, hepatotoxicity, developmental toxicity, and reproductive toxicity in animals. However, there is little information regarding the toxic effects of PFNA on oocyte maturation. In this study, we investigated the toxic effects of PFNA exposure on mouse oocyte maturation in vitro. Our results showed that 600 μM PFNA significantly inhibited germinal vesicle breakdown (GVBD) and polar body extrusion (PBE) in mouse oocytes. Our further study revealed that PFNA induced abnormal metaphase I (MI) spindle assembly, evidenced by malformed spindles and mislocalization of p-ERK1/2 in PFNA-treated oocytes. We also found that PFNA induced abnormal mitochondrial distribution and increased mitochondrial membrane potential. Consequently, PFNA increased reactive oxygen species (ROS) levels, leading to oxidative stress, DNA damage, and eventually early-stage apoptosis in oocytes. In addition, after 14 h culture, PFNA disrupted the formation of metaphase II (MII) spindle in most PFNA-treated oocytes with polar bodies. Collectively, our results indicate that PFNA interferes with oocyte maturation in vitro via disrupting spindle assembly, damaging mitochondrial functions, and inducing oxidative stress, DNA damage, and early-stage apoptosis.

Human oocyte maturation in vitro is improved by co-culture with cumulus cells from mature oocytes

2018 ◽  
Vol 36 (5) ◽  
pp. 508-523 ◽  
Author(s):  
Irma Virant-Klun ◽  
Chris Bauer ◽  
Anders Ståhlberg ◽  
Mikael Kubista ◽  
Thomas Skutella
Keyword(s):  
Oocyte Maturation ◽  
Cumulus Cells ◽  
Human Oocyte ◽  
Maturation In Vitro

338 EFFECTS OF CUMULUS CELLS AND FOLLICULAR FLUID ON PLASMINOGEN ACTIVATOR ACTIVITY AND OOCYTE MATURATION IN VITRO IN THE PIG

2006 ◽  
Vol 18 (2) ◽  
pp. 276
Author(s):  
C.-K. Park ◽  
J.-Y. An ◽  
S.-J. Sa ◽  
H.-T. Cheong ◽  
B.-K. Yang ◽  
...  
Keyword(s):  
Conditioned Medium ◽  
Oocyte Maturation ◽  
Follicular Fluid ◽  
Sodium Dodecyl ◽  
Cumulus Cells ◽  
The Other ◽  
Tissue Type ◽  
Bromophenol Blue ◽  
Maturation In Vitro

Plasminogen activators (PAs) are serine proteases, known to be secreted by a large number of cell type. PAs are reported to play a role in variety of physiologic processes, including fibrinolysis, ovulation, mammary involution, implantation, and fertilization. The present study investigated the effects of cumulus cells and porcine follicular fluid (pFF) on PA activity and oocyte maturation in vitro in the pig. Porcine oocytes were harvested from slaughterhouse ovaries, selected, and matured in modified North Carolina State University-23 (NCSU-23) media. After culture, cumulus-oocyte complexes (COCs) and denuded oocytes (DOs) were separately put into microtubes containing 20 �L of sample buffer [5.0% (w:v) sodium dodecyl sulfate, 20% (v:v) glycerol, and 0.0025% (w:v) bromophenol blue in 0.125 M Tris-HCl buffer] and frozen at -80�C until used for zymographic analysis. Differences in data were evaluated by Duncan's multiple-range test using the General Linear Models procedure in the Statistical Analysis System (SAS Institue, Inc., Cary, NC, USA). To determine the effect of porcine follicular fluid (pFF) on PA activity in porcine oocytes during maturation, the COCs and DOs were incubated in NCSU-23 medium with or without 10% (v/v) pFF for 0, 24, or 48 h. In the presence of cumulus cells, the proportions of oocytes matured to metaphase-II stage were significantly (P < 0.05) higher in medium with pFF than without pFF (69.8% vs. 37.7%, respectively). When COCs and DOs were cultured in the presence of pFF, tissue-type PA (tPA), urokinase-type PA (uPA), and tPA-PA inhibitor (tPA-PAI) were observed in COCs, and PA activities were higher at 48 h than 24 h. However, no PA activity was detected in DOs. Under the same conditions, when COCs and DOs were cultured in the absence of pFF, tPA and tPA-PAI were observed in COCs, and PA activities were increased as duration of culture increased. However, no PA activity was detected in DOs. When porcine oocytes were cultured in the presence of pFF, the activities of tPA-PAI, tPA, and uPA were observed in conditioned medium with COCs and DOs cultured for 24 h and 48 h. In the absence of pFF, PA activities were observed only in conditioned medium with COCs, and no PA activities were detected in conditioned medium with DOs. On the other hand, three plasminogen-dependent lytic bands (tPA-PAI, tPA, and uPA) were observed in pFF cultures. Particularly uPA activity was higher than the other kinds of PA activity. When oocytes and cumulus cells were separated from porcine COCs at 0 h of cultrue, tPA-PAI, tPA, and uPA were detected in cumulus cells at 48-h culture, but no PA activities were in DOs. The presence of pFF and cumulus cells in maturation medium stimulated not only nuclear and cytoplasmic maturation in porcine COCs, but also PA production by cumulus cells and COCs. It is possible that PAs produced by cumulus cells migrated through the gap junction between oocyte and cumulus cells. These results suggest that porcine oocytes have no ability to produce PA themselves.

127 Temporal regulation of molecular pathways, metabolic enzymes and growth factor receptors during bovine oocyte maturation in vitro

2019 ◽  
Vol 31 (1) ◽  
pp. 189
Author(s):  
S. Rajput ◽  
J. Becker ◽  
Y. Yuan ◽  
W. Schoolcraft ◽  
R. Krisher
Keyword(s):  
Growth Factor ◽  
Oocyte Maturation ◽  
Growth Factor Receptors ◽  
Cumulus Cells ◽  
Metabolic Enzymes ◽  
Molecular Pathways ◽  
Temporal Regulation ◽  
Maturation In Vitro ◽  
Significant Change

Although great efforts have been made to improve in vitro oocyte maturation (IVM) medium, we have yet to achieve competence equivalent to in vivo-matured oocytes. The failure in development of culture conditions for IVM yielding high quality eggs is attributed to an incomplete understanding of molecular pathways regulating oocyte and cumulus cell metabolism. The objective of the present study was to characterise the expression and functional activity of cell signalling pathways (mTOR, AKT, 4EBP1, ERK1/2), metabolic enzymes (PKM2, PDH, LDHA, AMPK), and growth factor receptors (IGF1R, IGFIIR, EGFR, FGFR1) in bovine oocytes and cumulus cells before and after in vitro maturation. In vitro-derived cumulus-oocyte complexes were collected at germinal vesicle (GV) and metaphase II (MII) stages (20 cumulus-oocyte complexes per stage; n=3 replicates) and subjected separately to Western blot analysis using antibodies against both phosphorylated (p) and total (t) protein abundance; the ratio p:t was used to determine the activity of each pathway. Results demonstrate increased (P&lt;0.05) mTOR and ERK1/2 signalling, with no change in AKT and 4EBP1 activity, in oocytes during IVM. We observed increased (P&lt;0.05) abundance of oocyte t-ERK from the GV to MII stage, but total expression of AKT, mTOR and 4EBP1 did not change. In cumulus cells, there was a significant (P&lt;0.05) reduction in mTOR and 4EBP1, an increase in AKT, and no significant change in ERK activity. Analysis of metabolic enzymes in oocytes demonstrated increased (P&lt;0.05) PDH, reduced AMPK, and unchanged PKM2 and LDHA phosphorylation during IVM. However, increased expression of t-PKM2 abundance was observed from the GV to MII stage. In cumulus cells, tAMPK abundance was reduced (P&gt;0.05), but no significant change was observed in the activity of other metabolic enzymes analysed during IVM. Finally, we observed abundant expression of IGF2R in the oocyte compared with other growth factor receptors analysed, although IGF2R was significantly (P&lt;0.05) reduced from GV to MII oocytes. In cumulus cells, both IGF1R and IGF2R were highly abundant compared with EGFR and FGFR but did not change during IVM. Data were analysed using one-way ANOVA. Results suggest that regulatory mechanisms including AKT/mTOR/4EBP1 and ERK are entirely different in oocytes and cumulus cells during maturation. An increase in the inhibitory phosphorylation of oocyte PDH (S293) toward the end of maturation suggests low metabolism of pyruvate via the Krebs cycle at that time. Similarly, dephosphorylation of AMPK (T172) suggests reduced AMPK activity and reduced fatty acid oxidation in mature oocytes. In addition, temporal regulation of IGF1R in the oocyte and EGFR in cumulus cells suggests an important role for these growth factor receptors during maturation and that these growth factors could be used to improve IVM medium in the bovine. Collectively, these results increase our understanding of the molecular pathways regulating oocyte metabolism during maturation and provide a strategy to improve the IVM environment for assisted reproductive technology.

scholarly journals Effects of steroids on mouse oocyte maturation in vitro

Reproduction ◽  
1980 ◽  
Vol 60 (2) ◽  
pp. 331-338 ◽  
Author(s):  
D. M. Smith ◽  
D. Y. Tenney
Keyword(s):  
Oocyte Maturation ◽  
Mouse Oocyte ◽  
Maturation In Vitro

scholarly journals The effect of cold shock on mouse oocyte maturation in vitro

Reproduction ◽  
1978 ◽  
Vol 54 (2) ◽  
pp. 401-403
Author(s):  
D. M. Smith ◽  
D. Y. Tenney
Keyword(s):  
Oocyte Maturation ◽  
Cold Shock ◽  
Mouse Oocyte ◽  
Maturation In Vitro
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