scholarly journals Tingkat Kematangan Inti Oosit Sapi Setelah 24 Jam Presevasi Ovarium

2015 ◽  
Vol 15 (2) ◽  
pp. 72-78
Author(s):  
Rini Widyastuti ◽  
Siti Darodjah Rasad
Keyword(s):  
Phosphate Buffer ◽  
Bovine Oocyte ◽  
Oocyte Maturity ◽  
Phosphate Buffer Saline ◽  
Bovine Oocytes ◽  
Meiotic Competence

(Nuclear maturity of bovine oocyte after 24 hours ovary preservation)ABSTRACT. The objective of the research was to investigate  their meiotic competence or nuclear maturity of bovine oocytes maturity in vitro after 24 hour preservation on 5°C. Oocytes  were collected by slicing the ovaries in modified phosphate buffer saline (m-PBS). Selected cumulus-oocyte complexes (COCs) homogenous ooplasm  were cultured in maturity medium at 38°C in humidified atmosphere of 5% CO2 incubator. After 24 hours, oocytes stained for nuclear maturity’s evaluation. The proportion of oocytes at metaphase II (MII) was significantly difference (P 0.05) on oocytes that 24 hours preservated   (44.21 ± 3.04%) vs oocytes from fresh ovary (73.97 ± 9.32%) (P0.05). These results indicated that 24 hour’s preservation  bovine’s ovary on 50C  cause  decreases of nuclear oocyte maturity.

H1foo is essential for in vitro meiotic maturation of bovine oocytes

Zygote ◽  
2014 ◽  
Vol 23 (3) ◽  
pp. 416-425 ◽  
Author(s):  
Yan Yun ◽  
Peng An ◽  
Jing Ning ◽  
Gui-Ming Zhao ◽  
Wen-Lin Yang ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Polar Body ◽  
Linker Histone ◽  
Meiotic Maturation ◽  
Control Group ◽  
Bovine Oocyte ◽  
First Polar Body ◽  
Effective Sirnas ◽  
Bovine Oocytes

SummaryOocyte-specific linker histone, H1foo, is localized on the oocyte chromosomes during the process of meiotic maturation, and is essential for mouse oocyte maturation. Bovine H1foo has been identified, and its expression profile throughout oocyte maturation and early embryo development has been established. However, it has not been confirmed if H1foo is indispensable during bovine oocyte maturation. Effective siRNAs against H1foo were screened in HeLa cells, and then siRNA was microinjected into bovine oocytes to down-regulate H1foo expression. H1foo overexpression was achieved via mRNA injection. Reverse transcription polymerase chain reaction (RT-PCR) results indicated that H1foo was up-regulated by 200% and down-regulated by 70%. Based on the first polar body extrusion (PB1E) rate, H1foo overexpression apparently promoted meiotic progression. The knockdown of H1foo significantly impaired bovine oocyte maturation compared with H1foo overexpression and control groups (H1foo overexpression = 88.7%, H1foo siRNA = 41.2%, control = 71.2%; P < 0.05). This decrease can be rescued by co-injection of a modified H1foo mRNA that has escaped from the siRNA target. However, the H1e (somatic linker histone) overexpression had no effect on PB1E rate when compared with the control group. Therefore we concluded that H1foo is essential for bovine oocyte maturation and its overexpression stimulates the process.

scholarly journals 81 EFFECT OF SUCROSE CONCENTRATION IN WARMING MEDIUM ON THE DEVELOPMENT POTENTIAL OF VITRIFIED BOVINE OOCYTES

2005 ◽  
Vol 17 (2) ◽  
pp. 190
Author(s):  
W.C. Chang ◽  
J. Xu ◽  
S. Jiang ◽  
X.C. Tian ◽  
X. Yang ◽  
...  
Keyword(s):  
Sucrose Concentration ◽  
Control Group ◽  
Bovine Oocyte ◽  
Cell Stage ◽  
Significant Difference ◽  
Humidified Air ◽  
Bovine Oocytes ◽  
Experimental Group ◽  
One Way Anova

The aim of this experiment was to determine the effect of the sucrose concentration (0 to 0.33 M) in the dilution medium on the viability, fertilizability, and development of vitrified bovine oocytes. Bovine oocyte-cumulus complexes were collected from slaughterhouse ovaries and in vitro-matured as reported previously. After 24-h maturation in TCM199-based medium under 5% CO2 humidified air at 39°C, these were exposed to hyaluronidase and carefully pipetted to remove all except the 3–5 innermost layers of cumulus. Oocytes were put into the pre-equilibration medium for 3 min and then into vitrification solution containing HEPES-buffered TCM199 supplemented with 20% FBS, ethylene glycol, and dimethylsulphoxide for 25–30 s; they were then vitrified by modified solid surface vitrification (Dinnyes et al. 2000 Biol. Reprod. 63, 513–518).The oocytes were warmed at 39°C by placing them in holding medium with 0, 0.08, 0.17, 0.25, or 0.33 M sucrose. Non-vitrified oocytes were used as controls. Oocytes were inseminated 30 min after warming, and the presumptive zygotes were cultured in CR1-aa medium supplemented with 6 mg/mL BSA at 39°C in a humidified atmosphere of 5% CO2, 5% O2, and 90% N2 for eight days. Data were analyzed by one-way ANOVA. As shown in Table 1, there was no significant difference in survival rate (P > 0.05) of the vitrified oocytes that were placed in dilution solution containing 0.17, 0.25, or 0.33 M sucrose and the non-treated controls. On Day 2 (fertilized on Day 0), cleavage to the 8-cell stage was similar for the 0.17, 0.25, and 0.33 M dilution groups, but the rates for all three were significantly lower (P < 0.05) than for the control group. The blastocyst rate on Day 8 was significantly higher for the 0.25 M group than for any other experimental group but still significantly lower than for the control. In conclusion, this study suggests that with this vitrification/warming procedure the optimum concentration of sucrose in the dilution solution is 0.25 M. Table 1. Oocyte survival after vitrification/warming and subsequent embryo development The authors would like to thank Ms Colleen Shaffer for the preparation of bovine oocytes.

167 INHIBITION OF HEAT SHOCK PROTEIN 90 REDUCES DEVELOPMENTAL COMPETENCE OF BOVINE OOCYTES

2014 ◽  
Vol 26 (1) ◽  
pp. 197
Author(s):  
E. D. Souza ◽  
F. B. E. Paula ◽  
C. C. R. Quintao ◽  
J. H. M. Viana ◽  
L. T. Iguma ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
In Vitro Maturation ◽  
Hsp90 Inhibitor ◽  
Stress Conditions ◽  
Developmental Competence ◽  
Bovine Oocyte ◽  
Cleavage Rate ◽  
Bovine Oocytes

The 90-kDa heat shock protein (HSP90) is a chaperone that is important for maintaing protein homeostasis under stress conditions. HSP90 seems also to be required for maturation of Xenopus oocytes (Fisher et al. 2000 EMBO J. 19, 1516) and first cleavage of mouse zygotes (Audouard et al. 2011 PloS One 6, e17109). This study aimed to evaluate the effect of inhibition of HSP90 by 17-(allylamino)-17-demethoxygeldanamycin (17AAG, Sigma St. Louis, MO, USA) during in vitro maturation (IVM) on bovine oocyte developmental competence. Immature cumulus–oocyte complexes (COC) were randomly allocated in 3 treatments during IVM: T0 (control; n = 240), no HSP90 inhibitor; T1: 2 μM HSP90 inhibitor (17AAG; n = 250) for the first 12 h of IVM; and T2: 2 μM HSP90 inhibitor (n = 188) for 24 h of IVM. In vitro maturation was performed in Nunc plates containing 400 μL of TCM-199 medium (Invitrogen, Carlsbad, CA, USA) supplemented with porcine FSH (Hertape Calier, Juatuba, Brazil) and 10% oestrus cow serum under 5% CO2, 95% humidity, and 38.5°C for 24 h. Oocytes were in vitro fertilized for 20 h and incubated under the same IVM conditions. Semen was processed by Percoll gradient (Nutricell, Campinas, Brazil) an IVF performed with 2 × 106 spermatozoa mL–1. Presumptive zygotes were completely denuded in a PBS solution with hyaluronidase and then cultured in wells with 500 μL of modified CR2aa medium supplemented with 2.5% fetal calf serum (Nutricell) in an incubator at 38.5°C under 5% CO2, 5% O2, 90% N2, and saturated humidity. Cleavage rate was evaluated 72 h post-fertilization and blastocyst rates were evaluated at Day 7 and Day 8. Data from 6 repetitions were analysed by generalized linear model procedure of SAS software (version 9.1; SAS Institute Inc., Cary, NC, USA), and means were compared by Student-Newman-Keuls test. Values are shown as mean ± s.e.m. There was a tendency (P = 0.08) for a lower cleavage rate in T2 (52.6 ± 5.8%) than in T0 (control; 74.2 ± 4.1%). Inhibition of HSP90 by 17AAG for 12 h and 24 h of IVM (T1 and T2, respectively) decreased blastocyst rates at Day 7 (20.4 ± 3.0% and 14.3 ± 2.6%, respectively; P < 0.01) and Day 8 (22.6 ± 4.1% and 16.9 ± 2.7%, respectively; P < 0.05) when compared with control (T0 = 31.8 ± 2.5% and 34.1 ± 2.9% for Day 7 and Day 8, respectively). In addition, the inhibition of HSP90 for 24 h decreased (P < 0.05) the proportion of hatched blastocysts at Day 8 (9.5 ± 5.0% for T2, respectively) when compared with control (T0 = 35.8 ± 3.9%), indicating a reduction on embryo quality. In conclusion, inhibition of HSP90 by 17AAG during IVM results in lower developmental competence, suggesting that this protein is also important for bovine oocytes. Further studies are required to investigate if the role of HSP90 on developmental competence of bovine oocyte is affected when under stress conditions. The authors acknowledge CNPq 473484/2011-0, FAPEMIG and FAPES for financial support.

Effects of chronic exposure to 2G/3G cell phone radiationon in vitro maturation of bovine oocytes

2016 ◽  
Author(s):  
Yusuf Ziya Guzey ◽  
Ali Galip Onal
Keyword(s):  
Treatment Group ◽  
Mobile Phone ◽  
Cell Phone ◽  
In Vitro Maturation ◽  
Bovine Oocyte ◽  
Maturation Period ◽  
Mobile Phone Use ◽  
Fluorescent Stain ◽  
Bovine Oocytes

Mobile phone use has arisen and rapidly been increased during last decades and therefore electromagnetic radiation or heat sourcing from mobile phones and its possible damages has become focus of interest. The aim of the present study was to investigate the effects of mobile phone signals on in vitro maturation of bovine oocytes. A total of 224 bovine cumulus oocyte complexes were matured in vitro and separated into two groups, and only treatment group received mobile phone signals for 100 min by calling 5 min/h. Following maturation period, oocytes were stained by fluorescent stain to assess maturation rates. The results showed that oocytes reaching Metaphase I and Metaphase II stage were 35.2±2.24 % and 48.2±2.21 % for treatment group whereas 11.9±1.45 % and 73.8±4.60 % for controls, respectively (P<0.01). In conclusion, bovine oocyte maturation has been significantly reduced by mobile phone signals.

scholarly journals Investigation of surface topography of different root-end filling materials: An in vitro study

2019 ◽  
Vol 7 (1) ◽  
Author(s):  
Mine Koruyucu ◽  
Hazal Özcan ◽  
Merve Bayram ◽  
Abdullah Burak Cankaya ◽  
Nurullah Keklikoglu ◽  
...  
Keyword(s):  
Surface Topography ◽  
Normal Saline ◽  
Phosphate Buffer ◽  
Saline Solution ◽  
In Vitro Study ◽  
Phosphate Buffer Saline ◽  
Filling Materials ◽  
Retrograde Filling ◽  
Group B

Aim: Although there are many materials that can be used for retrograde filling in surgical endodontics, none of them can be regarded as an ideal material yet. The purpose of this study was to compare the surface topography of three different root-end filling materials.Methods: 36 extracted single rooted human incisor teeth were cleaned and decoronated to standardized 10 mm root lengths. The root segments were prepared and 2 mm apical resection were performed. The samples were randomly separeted to three groups (Group A: Ca(OH)2, Group B: MTA Angelus, Group C: ProRoot MTA), each comprised of 12 roots. Materials were placed as 2 mm apical barriers and obturated with guttapercha and AH-Plus sealer. Each group dimidiated two subgroups (A1,A2,B1,B2,C1,C2). Groups A1,B1,C1 were stored in normal saline (NS), groups A2,B2,C2 were stored in neutral phosphate buffer saline (NPBS) solution and samples were incubated at 370C for 2 weeks. Stereomicroscope (32X) was used to photograph the root-end filling.Results: All specimens demonstrated white crystals formation and sediment over the root-end filling materials and on the superficial border of the root-end cavities’ wall as a white plague. A2,B2,C2 samples have more crystal sediment on root-end fillings than samples A1,B1,C1. Dissolution and corrosion were observed in groups A1, A2.Conclusions: The results of this study revealed that calcium hydroxide is more resorbable than MTA Angelus and ProRoot MTA. The crystals formation and precipitation were observed in neutral phosphate buffer saline solution was more than normal saline solution for all groups as a hydroxiapatite crystals.  

Role of Intercellular Coupling on Chromatin Changes Transcriptional Activity and Meiotic Competence Acquisition During Bovine Oocyte Growth In Vitro.

2009 ◽  
Vol 81 (Suppl_1) ◽  
pp. 282-282
Author(s):  
Federica M. Franciosi ◽  
Valentina Lodde ◽  
Silvia Modina ◽  
Irene Tessaro ◽  
Alberto M. Luciano
Keyword(s):  
Transcriptional Activity ◽  
Bovine Oocyte ◽  
Intercellular Coupling ◽  
Oocyte Growth ◽  
Competence Acquisition ◽  
Meiotic Competence

Effect of α-tocopherol on in vitro maturation of bovine cumulus-oocyte complexes

2008 ◽  
Vol 88 (3) ◽  
pp. 463-467 ◽  
Author(s):  
A. Marques ◽  
P. Santos ◽  
G. Antunes ◽  
A. Chaveiro ◽  
F. Moreira da Silva
Keyword(s):  
Granulosa Cells ◽  
In Vitro Maturation ◽  
Detrimental Effect ◽  
Cumulus Cells ◽  
Bovine Oocyte ◽  
Slight Improvement ◽  
Maturation In Vitro ◽  
Bovine Oocytes ◽  
Cells Cultured

This study determined: the effects of α-tocopherol on apoptotic and necrotic levels of cumulus cells after in vitro maturation of bovine oocytes; whether exposure to α-tocopherol facilitates the development of bovine enclosed oocytes to metaphase II; and the effects of this antioxidant on apoptotic and necrotic levels of granulosa cells cultured in vitro. In conclusion, supplementation with α-tocopherol on in vitro maturation of bovine oocytes has a detrimental effect on the ability of oocytes to reach metaphase II, increasing the number of apoptotic and necrotic cumulus cells of bovine cumulus oocyte complexes (COC). This antioxidant showed a slight improvement in the viability of cultured granulosa cells at a concentration of 100 µM. Key words: Bovine, oocyte maturation in vitro, antioxidant, α-tocopherol

scholarly journals 94MEIOSIS INHIBITION OF BOVINE OOCYTES: EFFECTS ON SURVIVAL AFTER VITRIFICATION BY OPEN PULLED-STRAW METHOD

2004 ◽  
Vol 16 (2) ◽  
pp. 168
Author(s):  
C. Diez ◽  
M. Carbajo ◽  
L. Fernandez ◽  
C.O. Hidalgo ◽  
S. de la Varga ◽  
...  
Keyword(s):  
In Vitro Maturation ◽  
Cumulus Cells ◽  
Cell Types ◽  
Blastocyst Stage ◽  
Developmental Competence ◽  
Bovine Oocyte ◽  
17Β Estradiol ◽  
Bovine Oocytes ◽  
Range Test

Mammalian oocytes remain one of the most difficult cell types to successfully cryopreserve. The in vitro-maturation protocols (IVM) have a large impact on the oocyte maturation. Consequently, inhibition of meiosis has been used to improve developmental competence of oocytes without reducing blastocyst rates. Moreover, the meiotic stage influences the ability of oocytes to survive cryopreservation. This work analyzes the effect of the inhibition of meiosis (prematuration) on the freezability of the bovine oocyte. Cumulus-oocyte complexes (COCs) were recovered from slaughterhouse ovaries. Inmature oocytes (I) with compact cumulus and evenly granulated cytoplasm were selected. Prematuration (PM) was performed by incubating COCs for 22h in TCM199 NaHCO3 and roscovitine 25μM. IVM was accomplished in TCM199 NaHCO3, 10% FCS, FSH-LH and 17β-estradiol. Oocytes were subjected to 5 treatments prior the vitrification (see table). COCs were partially denuded from cumulus cells and vitrified/warmed using the OPS system (Vajta et al. 1998 Mol. Reprod. Dev. 51, 53–58). Warmed oocytes were fertilized (Day 0) and presumptive zygotes having a normal morphological appearance were cultured in SOFaa+5% of FCS (Day 3); elements with degenerated appearance were discarded and recorded. Fresh oocytes submitted to IVM (c-M) or prematured and matured (c-PM+M) were fertilized and cultured as controls. Data were analyzed by ANOVA and Duncan’s multiple range test and expressed as LSM±SE. Developmental data are referred to the zygotes cultured. Only oocytes vitrified after IVM reached the blastocyst stage, but at lower rates than fresh controls. However, no differences were found between treatments at any developmental stage. Oocytes vitrified both as prematured+matured and immature oocytes showed increased proportions (P&lt;0.01) of degenerated oocytes (37.3±5.9 and 49.9±5.9, respectively), as compared with oocytes matured before vitrification (17.6±5.9). These results show that effects induced by incubation with roscovitine (Lonergan et al. 2003 Mol. Reprod. Dev. 64, 369–378) in combination with cryodamage compromise the oocyte developmental ability. Supported by CICYT, AGL2001-379.

280 EFFECTS OF HOT SEASON ON BOVINE OOCYTE QUALITY: HOW TO BYPASS THE POOR OOCYTE QUALITY DURING THIS SEASON?

2015 ◽  
Vol 27 (1) ◽  
pp. 229
Author(s):  
L. Boccia ◽  
E. Iacono ◽  
B. Rossi ◽  
B. Merlo
Keyword(s):  
Degradation Products ◽  
Germinal Vesicle ◽  
Oocyte Quality ◽  
Female Reproductive Tract ◽  
Bovine Oocyte ◽  
Germinal Vesicle Breakdown ◽  
Serum Replacement ◽  
Hot Season ◽  
Meiotic Competence

Many authors attribute the decline of reproductive activity in summer to the heat stress, a multifactorial problem in which hyperthermia affects cellular function in various tissues of the female reproductive tract (Hansen et al. 2001; De Rensis et al. 2003). In particular, the combination of high temperatures and high humidity for a long period causes a reduced blood flow to uterus, oviducts, and ovaries, leading to a rise in the concentration of the degradation products of cellular activity. Therefore, the aim of this work was to elucidate the negative effect of the hot season on bovine oocyte quality and evaluate the influence of different factors on the acquisition of meiotic competence. In particular, meiotic competence of bovine oocytes recovered from animals housed at 44°28′00″ N, 11°26′00″ E during spring (March, 4–13°C) and summer (June, 16–27°C) was evaluated. Likewise, in summer the effect of an antioxidant, myo-inositol, the use of serum replacement (SR), and the use of oocytes recovered from cycling heifers (16–18 months) as compared to cows (>24 months) were tested. A total of 1346 abattoir-derived oocytes, equally divided for different experimental groups (over 6 replicates), were in vitro matured in TCM 199 supplemented with EGF (25 ng mL–1), IGF1 (100 ng mL–1), ITS supplement, pFSH-LH (0.1 IU each), and 10% FBS. Myoinositol was added at a concentration of 0, 15, 30, and 50 mM, while 10% SR was used alternatively to FBS. At the end of maturation period (20–22 h), oocytes were denuded and stained with 10 μg mL–1 of Hoechst 33342 at room temperature in the dark. After 15 min they were mounted on glass slides for evaluation of nuclear status using a Nikon Eclipse E400 microscope equipped with fluorescence filters. Nuclear configurations were classified as (a) germinal vesicle (GV), (b) germinal vesicle breakdown (GVBD), (c) metaphase I (M-I), (d) metaphase II (M-II), and (e) degenerated (DEG). Data are expressed as mean ± s.e.m. and were analysed by ANOVA (IBM SPSS Statistics) considering significance at P < 0.05. Oocyte quality of summer oocytes was significantly lower than spring counterparts as result of a higher rate of DEG (8.2 ± 0.6 v. 0.7 ± 0.6) and GV (5.4 ± 0.3 v. 0.4 ± 0.4, respectively; P < 0.05). Myo-inositol supplementation in IVM medium did not significantly affect either oocyte quality or meiotic competence in the hot season, such as the use of SR. When the oocytes were collected from cycling heifers ovaries during summer, the recovery rate of COC/ovary was significantly higher as compared to cows (4.5 v. 2.0), and a lower rate of DEG (1.8 ± 0.2; 8.2 ± 0.6) and GVBD (0.9 ± 0.6; 6.1 ± 0.3) was found (P < 0.05), even if the rate of GV (22.4 ± 0.1 v. 5.4 ± 0.3) was higher (P < 0.05) compared with cow. In conclusion, the hot season negatively affects oocyte quality, myo-inositol does not affect nuclear maturation, and SR can be used alternatively to FBS. The lower age of oocyte donor positively influenced the number of recoverable oocyte and degeneration rate.

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