• Title, Summary, Keyword: cumulus cell-oocyte complexes

Search Result 52, Processing Time 0.045 seconds

Effect of Cordycepin on the Cumulus Expansion and Meiotic Maturation of Mouse Cumulus-oocyte Complexes in Vitro (Cordycepin이 생쥐의 난구세포 분산과 난자의 성숙에 미치는 영향)

  • Lee, Gy-Soog
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.13 no.2
    • /
    • pp.201-206
    • /
    • 1986
  • These experiments were conducted to know whether RNA syntheis is involved in the cumulus expansion and oocyte maturation of mouse cumulus-oocyte complexes in vitro. Mouse cumulus-oocyte complexes(COC's) were cultured in the presence of cordycepin, an inhibitior of RNA synthesis and its effect on the cumulus expansion and oocyte maturation were examined. The results were as follows. 1. Continuous presence of cordycepin in the medium(200${\mu}g/ml$) inhibited the HCG-induced cumulus cell expansion of mouse complexes. This inhibition was reversible. 2. When the COC'S were preincubated with different concentration of cordycepin plus HCG for 3 hours and then transferred to the plain medium, the HCG induced cumulus expansion was suppressed at $100{\mu}g/ml$ of cordycepin. 3. When the COC'S were cultured with cordycepin after HCG stimulation(3hrs), the cumulus expansion were not suppressed by cordycepin. 4. Oocyte meiotic maturation did not appear to be affected by cordycepin. The data presented here imply that RNA synthesis is involved in the cumulus expansion process and that mouse oocytes are more resistant to RNA synthesis inhibitor than cumulus cells.

  • PDF

Cumulus Expansion and Oocyte Maturation of Oocyte-Cumulus Complexes Isolated from Different Ages of Mice In vitro (생쥐 Age에 따른 난자-난구 복합체의 분산과 성숙에 관한 연구)

  • Lee, Won-Kyo;Kwon, Hyuk-Bang
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.13 no.2
    • /
    • pp.195-200
    • /
    • 1986
  • In order to know when the cumulus cells of mouse follicles get ability to expand in vitro, the oocyte cumulus complexes obtained from different growing ages of mice were cultured in the medium containing HCG and their rate of expansion were observed and at the same time their maturation rate was examined. The growth of follicles was also checked by histological method. It was impossible to isolate the oocyte-cumulus complexes from 13 or 15 days old mouse ovaries. The oocyte-cumulus complexes collected from 17 days old mouse were partially induced to expanded by HCG, and from 19 days, most of the complexes were induced to full expansion. The rate of cumulus cell expansion by HCG and the oocyte maturation increased steadly during the growing ages to adult. Thus, the time for follicles to get competence for expansion and maturation seems to be closely related. Antral follicles were appeared from 17 days old mice and Graafian follicles were seen from 21 days old mice. The competence for cumulus expansion increased during follicle growth up to 21 days old mice.

  • PDF

Effects of Magnetized Medium on In Vitro Maturation of Porcine Cumulus Cell-Oocyte Complexes

  • Kim, Yun-Jung;Lee, Sang-Hee;Jung, Soo-Jung;Park, Choon-Keun
    • Journal of Magnetics
    • /
    • v.19 no.3
    • /
    • pp.241-247
    • /
    • 2014
  • The objective of this study was to study the effect of magnetized water on porcine cumulus cell-oocyte complexes (COCs). Oocytes obtained from female pig were cultured in a medium magnetized at 0, 2000, 4000, and 6000 Gauss (G) for 5 minutes using the neodymium magnet. Subsequently, intracellular hydrogen peroxide ($H_2O_2$) concentration, glutathione (GSH) activity, oocyte membrane integrity, anti-apoptosis factor Bcl-xL expression, and nuclear maturation were analyzed. The intracellular $H_2O_2$ levels in COCs cultured for 44 hours were not significantly different among the variously magnetized samples. However, GSH activity were significantly higher in the magnetized samples compared to the 0 G sample. The Bcl-xL mRNA expression in COCs cultured for 44 hours was higher in the 4000 G sample than other treatment groups. Membrane damage in COCs cultured for 22 and 44 hours was significantly lower in 4000 G group than control group. On the other hand, nuclear stages as maturation indicator significantly increased in 2000, 4000, and 6000 G groups compared to 0 G group. These results indicate that incubation of porcine oocytes and cumulus cells in magnetized medium improves intracellular GSH levels, membrane integrity and nuclear maturation, and inhibits apoptosis in vitro.

Studies on the Metabolic Cooperativity between Ooccte and Cumulus Cells in Mammalian Oocyte Cumulus Complexes in vitro (포유동물 난자-난구 복합체의 Metabolic cooperativity)

  • 고선근;나철호;권혁방
    • The Korean Journal of Zoology
    • /
    • v.31 no.2
    • /
    • pp.81-86
    • /
    • 1988
  • The relationship between cumulus cell expansion, cocyte maturation and metabolic cooperativitiy was investigated by using mouse and pig cocyte-cumulus complexes in vitro. Cocyte germinal vesicle breakdown (GVBD) and cumulus expansion were manipulated with hormones or reagents which increase intracellular cAMP leveL Metabolic cooperativity between oocyte and cumulus cells was assessed by determination of the fraction of radiolabelled uridine marker that was transferred from the cumulus mass to the oocyte. Uptake of uddine marker by mouse and pig cumulus mass was increased by about fourfold of basal level with the stimulation of hormones (human choriononic gonadotrophin, HCG; follicle stimulating hormone, FSH) or cyclic AMP sttmulators (3-isobutyl-1-methylxanthine, IBMX; forskolin) during culture. However, the fraction of uridine that was transferred from the cumulus mass to the cocyte (transfer ratio) was gradually decreased during culture, irrespective with the presence of hormones or stimulators. The decrease of the transfer ratio was not correlated with the state of occyte whether they have GV or not, or with the degree of cumulus expansion. In mouse complexes, HCG induced more significant reducton of transfer ratio than other treatments. These results do not support the idea that modulations of metabolic cooperativity between cumulus cells and oocytes are important for the regulation of meiotic resumption in mammals.

  • PDF

Effect of Human Cord Serum on Oocyte Maturation and Cumulus Cell Expansion (신생아제대혈청이 난자성숙과 난구세포 분산에 미치는 영향)

  • Lee, Yu-Il;Park, Hyun-Jeong;Kwon, Young-Suk
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.25 no.1
    • /
    • pp.9-16
    • /
    • 1998
  • This study was performed to investigate the stimulating effect on oocyte maturation and cumulus cell expansion in TC199 media by human cord serum (HCS) supplementation. Immature mouse oocyte cumulus complexes (OCCs) were cultured in TC199 media supplemented with bovine serum albumin (BSA), HCS and human chorionic gonadotropin (hCG) instead of luteinizing hormone (LH) respectively, and the expression of cumulus expansion and oocyte maturation were observed. After 4hr and 24hr culture with or without OCCs, media containing 0.4% BSA, 10% HCS and 10 IV hCG respectively were collected and analyzed for changing concentrations of estradiol $(E_2)$, progesterone $(P_4)$, testosterone (T), and $PGF_{2\alpha}$. There were no elevation of $E_2$, T, and $PGF_{2\alpha}$ by OCCs culture, but minute elevation of $P_4$ level by 24hr OCCs culture in hCG supplementation (p=0.048). The stimulating pattern of cumulus expansion of OCCs by HCS and hCG supplementation was similar to our previously report using Ham's F-10 media, however oocyte maturation rates after 24hr OCCs culture in all media were increased by $20\sim30%$ compared to Ham's F-10 media. These results suggest that LH in HCS induce cumulus expansion probably by $P_4$ secretion of OCCs, and TC199 is efficient media for immature mouse oocyte maturation.

  • PDF

Reversible Effects of Exogenous GM3 on Meiotic Maturation and Cumulus Cells Expansion of Porcine Cumulus-oocyte Complexes

  • Kim, Jin-Woo;Park, Hyo-Jin;Jung, Jae-Min;Yang, Seul-Gi;Kim, Min-Ji;Kim, In-Su;Jegal, Ho-Geun;Koo, Deog-Bon
    • Journal of Embryo Transfer
    • /
    • v.33 no.4
    • /
    • pp.287-296
    • /
    • 2018
  • Ganglioside GM3 is known as an inhibition factor of cell differentiation and proliferation via inhibition of epidermal growth factor receptor (EGFR) phosphorylation. Our previous study showed that the exogenous ganglioside GM3 reduced the meiotic maturation of porcine oocytes and induced apoptosis at 44 h of in vitro maturation (IVM). However, the role of ganglioside GM3 in the relationship between EGFR signaling and apoptosis during porcine oocyte maturation has not yet been studied. First, porcine cumulus-oocyte complexes (COCs) were cultured in the NCSU-23 medium with exogenous ganglioside GM3 according to maturation periods (non-treated, only IVM I: 0 - 22 h, only IVM II: 22 - 44 h and IVM I & II: 0 - 44 h). We confirmed that the proportion of germinal vesicle breakdown (GVBD) increased significantly in the IVM I treated group than in the control group. We also confirmed that the meiotic maturation until M II stage and polar body formation decreased significantly in the only IVM I treated group. Cumulus cell expansion and mRNA levels of the expansion-related factors (HAS2, TNFAIP6 and PTX3) decreased significantly in the IVM I treated group than in the control group. Protein levels of EGFR, p-EGFR, ERK1/2, and p-ERK1/2 decreased significantly in the GM3-treated groups, during the IVM I period. In addition, cellular apoptosis, determined using TUNEL assay, and protein levels of Cleaved caspase 3, were increased significantly in the GM3-treated COCs during the IVM I period. Based on these results, ganglioside GM3 exposure of porcine COCs during the IVM I period reduced meiotic maturation and cumulus cell expansion via inhibition of EGFR activity in pigs.

Porcine Oocytes with Meiotic Competence are Synchronized Early Stage of Germinal Vesicle (GV)

  • Park, Kwang-Wook
    • Reproductive and Developmental Biology
    • /
    • v.34 no.3
    • /
    • pp.169-173
    • /
    • 2010
  • Correlations between cumulus cells and germinal vesicle (GV) chromatin configuration were examined in porcine oocytes. Cumulus-oocyte complexes (COCs) were collected from 2~6 mm follicles and divided into three categories according to cumulus cell morphology. "A" group was compacted COCs with more than three cumulus cell layers. "B" group was COCs with less cumulus cell layers than "A" group. "C" group was COCs with one or less layer of cumulus cells. Cumulus cells were removed 0.1% hyaluronidase, and denuded oocytes were stained with Hoechst 33342. GV chromatin configuration was classified into GV-Con and GV-Dis. GV-Con meant that a nucleus was surrounded by condensed chromatin in a ring. GV-Dis meant that filamentous chromatin clumps were distributed in nucleus. The proportion (80.2%) of GV-Con in "A" group was significantly higher than "B" (62.0%) or "C" (44.9%). The proportion (55.1%) of GV-Dis in "C" group was significantly higher than "A" (19.8%) or "B" (38.0%). The meiotic competence of COCs was examined after 44 h culture. The proportion (90.0%) of oocytes reaching to metaphase II (M-II) in "A" group was significantly higher than "B" (76.5%) or "C" (45.5%). In conclusion, oocytes with good quality cumulus cell layers are synchronized early GV stage, and early GV stage is important for meiotic competence in pigs.

Gonadotropins Improve Porcine Oocyte Maturation and Embryo Development through Regulation of Maternal Gene Expression

  • Wang, Qing-Ling;Zhao, Ming-Hui;Jin, Yong-Xun;Kim, Nam-Hyung;Cui, Xiang-Shun
    • Journal of Embryo Transfer
    • /
    • v.28 no.4
    • /
    • pp.361-371
    • /
    • 2013
  • The present study assessed the effect of FSH and LH on oocyte meiotic, cytoplasmic maturation and on the expression level and polyadenylation status of several maternal genes. Cumulus-oocyte complexes were cultured in the presence of FSH, LH, or the combination of FSH and LH. Significant cumulus expansion and nuclear maturation was observed upon exposure to FSH alone and to the combination of FSH and LH. The combination of FSH and LH during entire IVM increased the mRNA level of four maternal genes, C-mos, Cyclin B1, Gdf9 and Bmp15, at 28 h. Supplemented with FSH or LH significantly enhanced the polyadenylation of Gdf9 and Bmp15; and altered the expression level of Gdf9 and Bmp15. Following parthenogenesis, the exposure of oocytes to combination of FSH and LH during IVM significantly increased cleavage rate, blastocyst formation rate and total cell number, and decreased apoptosis. In addition, FSH and LH down-regulated the autophagy gene Atg6 and upregulated the apoptosis gene Bcl-xL at the mRNA level in blastocysts. These data suggest that the FSH and LH enhance meiotic and cytoplasmic maturation, possibly through the regulation of maternal gene expression and polyadenylation. Overall, we show here that FSH and LH inhibit apoptosis and autophagy and improve parthenogenetic embryo competence and development.

Expression of $interferon$ $regulatory$ factor-1 in the mouse cumulus-oocyte complex is negatively related with oocyte maturation

  • Kim, Yun-Sun;Kim, Eun-Young;Moon, Ji-Sook;Yoon, Tae-Ki;Lee, Woo-Sik;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.38 no.4
    • /
    • pp.193-202
    • /
    • 2011
  • Objective: We found previously that $interferon$ $regulatory$ factor ($Irf$)-1 is a germinal vesicle (GV)-selective gene that highly expressed in GV as compared to metaphase II oocytes. To our knowledge, the function of $Irf-1$ in oocytes has yet to be examined. The present study was conducted to determine the relationship between retinoic acid (RA) and RA-mediated expression of $Irf-1$ and the mouse oocyte maturation. Methods: Immature cumulus-oocyte-complexes (COCs) were collected from 17-day-old female mice and cultured $in$ $vitro$ for 16 hours in the presence of varying concentrations of RA (0-10 ${\mu}M$). Rate of oocyte maturation and activation was measured. Gene expression was measured by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) and cytokine secretion in the medium was measured by Bio-Plex analysis. Apoptosis was analyzed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. Results: The rates of oocyte maturation to metaphase II and oocyte activation increased significantly with RA treatment (10 nM-1 ${\mu}M$). With 100 nM RA treatment, lowest level of $Irf-1$ mRNA and cumulus cell's apoptosis was found. Among 23 cytokines measured by Bio-Plex system, the substantial changes in secretion of tumor necrosis factor-${\alpha}$, macrophage inflammatory protein-$1{\beta}$, eotaxin and interleukin-12 (p40) from COCs in response to RA were detected. Conclusion: We concluded that the maturation of oocytes and $Irf-1$ expression are negatively correlated, and RA enhances the developmental competence of mouse immature oocytes $in$ $vitro$ by suppressing apoptosis of cumulus cells. Using a mouse model, results of the present study provide insights into improved culture conditions for $in$ $vitro$ oocyte maturation and relevant cytokine production and secretion in assisted reproductive technology.

Expression Analysis of Matrix Metalloproteinases and Tissue Inhibitor of Matrix Metalloproteinases from In Vitro Maturation Oocytes Complexes in Porcine (돼지 체외성숙난자에서 MMPs와 TIMPs의 발현 분석)

  • Kim, Sang-Hwan;Kang, Hyun-Ah;Kim, Dae-Seung;Lee, Myeong-Seop;Seo, Kang-Suk;Min, Kwan-Sik;Yoon, Jong-Taek
    • Reproductive and Developmental Biology
    • /
    • v.34 no.1
    • /
    • pp.55-62
    • /
    • 2010
  • Matrix metalloproteinases (MMP) play important roles in extracellular matrix (ECM) remodeling during ovarian follicular development, oocytes development and ovulation. In an attempt to investigate the effect of MMP activation in development cumulus-oocytes complexes, we examined the localization and expression of MMP, and monitored MMP expression profile. Cumulus-oocytes complexes were collected and matured in vitro for 24 hr, 36 hr and 48 hr. A mRNA expression of MMP-2, MMP-9, TIMP-2 and TIMP-3 was detected in all culture medium regardless of CC, DC and CDCs. Activity of MMP-2 in the DC progressively was increased from 24 hr to 48 hr. But MMP-9 was not detected in all culture medium. The localization of MMP-2 was also measured by immunohistochemistry analysis. The MMP-2 and TIMP-2 was detected in cumulus cell and oocyte zone pellucida. Expression of MMP-2 protein in the COCs was progressively increased from 24 hr to 48 hr. However, MMP-9 protein was progressively decreased from 24 hr to 48 hr. And TIMP-2 protein was most highly expressed in the CDCs 36 hr. Expression of TIMP-3 protein in the CDCs was progressively increased from 24 hr to 48 hr. In conclusion, these results suggest that MMP-2 plays a role in maintaining normal maturation and development by controlling the ECM inhibitor concentration on cumulus cell and oocytes.