scholarly journals 204.Regulation of bovine oocyte meiotic and developmental capacity by glucose and glucosamine

2004 ◽  
Vol 16 (9) ◽  
pp. 204
Author(s):  
M. L. Sutton McDowall ◽  
R. B. Gilchrist ◽  
J. G. Thompson
Keyword(s):  
Protein Synthesis ◽  
Follicular Fluid ◽  
Glucose Concentration ◽  
Defined Medium ◽  
Developmental Competence ◽  
Fluid Composition ◽  
Fluid Medium ◽  
Nuclear Maturation ◽  
Developmental Capacity

Glucose is an important substrate for in vitro oocyte maturation (IVM) and is metabolised by cumulus oocyte complexes (COCs) via glycolysis or is used for extracellular matrix (ECM) synthesis. Follicular glucose concentration is significantly lower than commonly used IVM media (2.3�mM v. 5.6�mM in TCM199). Glucosamine is an alternative substrate for ECM and supplementation to IVM media reduces glucose uptake by COCs. The aim of this study was to determine the effect of glucose and glucosamine supplementation during IVM on bovine oocytes. First, bovine COCs (n�=�400) were matured in TCM199 (containing pyruvate, BSA, hCG and FSH), or synthetic follicular fluid medium (SFFM; a defined medium based on bovine follicular fluid composition) with 2.3�mM or 5.6�mM glucose���5�mM glucosamine and nuclear maturation was assessed after 24 and 30�h. Significantly less COCs matured in 2.3�mM glucose completed nuclear maturation compared to COCs matured in 5.6�mM glucose (P�<�0.05), whereas glucosamine had no effect on meiotic maturation. We then compared oocyte developmental capacity following IVM (n�=�600) in TCM199 or SFFM�+�5.6�mM glucose���5mM glucosamine. Blastocyst production was severely perturbed when COCs were matured in the presence of glucosamine (–glucosamine 32% v. +glucosamine 4%; P�<�0.001). To determine the cause of this reduction in oocyte developmental competence, we investigated oocyte protein synthesis by maturing COCs (n�=�100) in SFFM�+�5.6�mM glucose���5mM glucosamine�+�1�mM L-[2,3,4,5,6–3H] phenylalanine. In the presence of glucosamine, oocyte protein synthesis was reduced 40% compared to oocytes matured in control medium (P�<�0.05). These results demonstrate that while glucosamine supplementation has no effect on oocyte nuclear maturation, cytoplasmic maturation is compromised, as demonstrated by perturbed oocyte protein synthesis and embryo development. In contrast, glucose concentration has a significant influence on meiotic progression. This provides a useful model to investigate the mechanisms of establishment of developmental competence in oocytes following maturation.

scholarly journals 300 GLUCOSAMINE SUPPLEMENTATION DURING IN VITRO MATURATION LEADS TO PERTURBED DEVELOPMENTAL CAPACITY OF BOVINE CUMULUS OOCYTE COMPLEXES

2005 ◽  
Vol 17 (2) ◽  
pp. 300 ◽  
Author(s):  
M. Sutton-McDowall ◽  
R. Gilchrist ◽  
J. Thompson
Keyword(s):  
Protein Synthesis ◽  
Follicular Fluid ◽  
Defined Medium ◽  
Signalling Pathway ◽  
Blastocyst Development ◽  
Fluid Medium ◽  
Nuclear Maturation ◽  
Developmental Capacity ◽  
Pi3k Signalling

Glucose is a primary energy substrate required for successful in vitro oocyte maturation (IVM). However, most maturation media contain more glucose than that seen in follicular fluid (2.3 mM vs. 5.6 mM in TCM199). Glucosamine (Glc) as an alternative substrate for extracellular matrix during cumulus expansion reduced glucose uptake by bovine cumulus oocyte complexes (COCs, Sutton-McDowall et al. 2004 Reproduction 128, 313–319). As this could enable a reduction in glucose concentrations to physiological levels in IVM medium of COCs, the aim of this study was to investigate the influence of Glc supplementation on oocyte developmental capacity. Bovine COCs were matured in synthetic follicular fluid medium (SFFM, a defined medium based on the composition of follicular fluid, plus 5.6 mM glucose, FSH, hCG and BSA, Sutton-McDowall et al. 2004 Reprod. Fertil. Dev. 16 sup, 204) ± 5 mM Glc. After 24 h, either nuclear maturation (rep = 8, n = 160) or blastocyst development 8 days post-fertilization (rep = 5, n = 400) was determined. Data was arcsine transformed and analyzed by ANOVA, followed by Tukey's test. While the presence of Glc did not affect the completion of nuclear maturation and early cleavage, +Glc led to severely perturbed blastocyst development (−Glc, 32.5 ± 1.9% vs. +Glc, 4.7 ± 3.9%, P < 0.001). Glc supplementation in somatic cells is well-known to down-regulate the phosphatidylinositol-3-kinase (PI3K) signalling pathway, reducing protein synthesis and other cell survival mechanisms. Therefore, oocyte protein synthesis (measured by [2,3,4,5,6-3H] phenylalanine incorporation, rep = 5, n = 200) and embryo development (rep = 6, n = 720) following IVM in SFFM ± Glc ± EGF (a PI3K pathway stimulator) was determined. Glc supplementation led to a 40% decrease in protein synthesis compared to −Glc, while the combination of +Glc + EGF significantly increased protein synthesis by 60%. However, IVM + EGF + Glc did not improve blastocyst rates (main effect: −Glc 41.6 ± 6.6% vs. +Glc, 6.6 ± 1.7%, P < 0.001). Additionally, COCs were also cultured in SFFM ± 50 μM LY294002 (a specific PI3K inhibitor) and nuclear maturation (rep = 5, n = 200) or blastocyst development 8 days post-fertilization (rep = 4, n = 200) was determined. Despite the presence of LY294002 leading to 43% less COCs completing nuclear maturation (P < 0.001), blastocyst development was not affected (mean = 38.8 ± 3.2%). These results demonstrate that Glc supplementation during IVM has no effect on nuclear maturation or early development but is detrimental to oocyte developmental capacity by severely perturbing blastocyst development. However, the diminished developmental capacity appears to be independent of the well-characterized Glc down-regulation of the PI3K signalling pathway. This work was supported by the Australian Research Council (SPIRT, C00107702) and Cook Australia Pty Ltd.

Improvement of the developmental competence of canine oocyte using caffeine supplementation during IVM at different maturation time

Zygote ◽  
2018 ◽  
Vol 26 (2) ◽  
pp. 162-167 ◽  
Author(s):  
Mohamed Fathi ◽  
A. Salama ◽  
Magdy R. Badr
Keyword(s):  
Developmental Competence ◽  
Control Group ◽  
Free Medium ◽  
Maturation Time ◽  
Fluid Medium ◽  
Developmental Potential ◽  
Nuclear Maturation ◽  
Maturation Medium ◽  
Oviductal Fluid

SummaryThe aim of the current study was to investigate the effect of caffeine supplementation during in vitro maturation (IVM) for different maturation times on the developmental potential of canine oocytes recovered from ovariohysterectomized bitches. The recovered cumulus–oocytes complexes were in vitro matured for 72 h. Here, 10 mM caffeine was added to the maturation medium for different incubation times (caffeine from 0–72 h maturation, caffeine for the first 24 h of maturation only, caffeine addition from 24 to 48 h maturation time, caffeine addition from 48 to 72 h maturation or in caffeine-free medium, control group). The matured oocytes were in vitro fertilized using frozen–thawed spermatozoa. The presumptive zygotes were in vitro cultured in synthetic oviductal fluid medium for 5 days. The results showed that both maturation and fertilization rates were significantly higher (P ˂ 0.05) using caffeine-treated medium for the first 24 h of maturation compared with the control and other two groups of caffeine treatment (from 24 to 48 h and from 48 to 72 h), whereas use of caffeine-treated medium for a 0–72 h incubation time did not affect these rates (P > 0.05). Interestingly, the matured oocytes in caffeine-supplemented medium for the first 24 h or from 0–72 h showed a significant (P ˂ 0.05) increase in the total number of cleaved embryos compared with the control group. In conclusion, supplementation of the maturation medium with 10 mM caffeine for the first 24 h of maturation or during the whole maturation time (0–72 h) improved nuclear maturation and subsequent embryo development preimplantation following in vitro fertilization.

206 EFFECT OF ADDITION OF FOLLICULAR FLUID OR GROWTH DIFFERENTIATION FACTOR-9 ON IN VITRO MATURATION OF PORCINE OOCYTES DENUDED 20h AFTER THE START OF IN VITRO MATURATION

2016 ◽  
Vol 28 (2) ◽  
pp. 234
Author(s):  
P. Ferré ◽  
T. T. M. Bui ◽  
M. T. Tran ◽  
T. Wakai ◽  
H. Funahashi
Keyword(s):  
Follicular Fluid ◽  
In Vitro Maturation ◽  
Cumulus Cells ◽  
Developmental Competence ◽  
Dapi Staining ◽  
Experimental Conditions ◽  
Nuclear Maturation ◽  
Differentiation Factor ◽  
Growth Differentiation Factor 9

The interruption of communication between oocyte and cumulus cells (CC) can trigger meiotic resumption and exogenous additives, such as follicular fluid (FF) and growth differentiation factor-9 (GDF9), can improve oocyte quality and the developmental competence. This study was undertaken to examine if the absence and presence of FF from medium follicles (MF; 3–6 mm in diameter) or recombinant human GDF9 (Biovision, Milpitas, CA, USA) during the first or/and second half of in vitro maturation (IVM) had any effects on IVM of oocytes from small follicles (SF; 0.5–2 mm in diameter) or MF when the oocytes were denuded at 20 h after the start of IVM. Cumulus-oocyte complexes (COC) were aspirated from SF or MF of slaughtered prepubertal gilt ovaries. Groups of ~30 COC were cultured in a 300-μL drop of porcine oocyte medium containing 50 µM β-mercaptoethanol (mPOM) with or without 10% (v/v) FF and/or 100 ng mL–1 GDF9 at 39°C and 5% CO2 in air. During the first 20 h after the start of IVM, the medium was supplemented with 1 mM dibutyryl c-AMP, 10 IU mL–1 eCG and 10 IU mL–1 hCG. After the first period of IVM, the CC surrounding the oocytes were removed and the denuded oocytes continued culture for IVM with or without FF or/and GDF9 in the absence of dibutyryl c-AMP and gonadotropins in the same medium for another 24 h. At the end of IVM, meiotic progression of the oocytes was examined by DAPI staining. Statistical analyses from at least 4 replicates data were performed by a 2-way ANOVA and a Tukey’s multiple comparisons test. Removal of CC 20 h after the start of IVM significantly improved the incidence of mature oocytes derived from SF (59.2–64.1% v. 41.6–43.1% in controls, P < 0.05) but not from MF (73.1–78.5% v. 70.6–71.8% in controls), whereas regardless of supplementation with FF or GDF9, the maturation rates were always significantly higher in the denuded oocytes from MF (72.4–83.6%) than SF (57.8–66.2%; P < 0.05). Despite of the origin of COC (SF or MF), maturation rates of oocytes denuded 20 h after the start of IVM were not affected by supplementation with FF or GDF9 during the first and/or second half of IVM (P > 0.05). In summary, CC removal from COC 20 h after the start of IVM promotes nuclear maturation of oocytes from SF. Exogenous additives such as GDF9 and follicular fluid from MF do not seem to affect the promotion of nuclear maturation in our experimental conditions.

scholarly journals Porcine follicular fluid derived from > 8 mm sized follicles improves oocyte maturation and embryo development during in vitro maturation of pigs

Zygote ◽  
2020 ◽  
pp. 1-6
Author(s):  
Ji-Eun Park ◽  
Sang-Hee Lee ◽  
Yong Hwangbo ◽  
Choon-Keun Park
Keyword(s):  
Embryonic Development ◽  
Follicular Fluid ◽  
In Vitro Maturation ◽  
Developmental Competence ◽  
Control Group ◽  
Cumulus Expansion ◽  
Treatment Groups ◽  
Nuclear Maturation ◽  
Cytoplasmic Maturation

Summary The aim of the present study was to investigate the effects of porcine follicular fluid (pFF) from large-sized (LFF; >8 mm in diameter) and medium-sized (MFF; 3–6 mm in diameter) follicles on the maturation and developmental competence of porcine oocytes. Cumulus–oocyte complexes (COCs) were collected from follicles 3–6 mm in diameter. The collected COCs were incubated for 22 h with LFF or MFF (in vitro maturation (IVM)-I stage) and were incubated subsequently for 22 h with LFF or MFF (IVM-II stage). Cumulus expansion was confirmed after the IVM-I stage and nuclear maturation was evaluated after the IVM-II stage. Intracellular glutathione (GSH) and reactive oxygen species (ROS) levels were measured and embryonic development was evaluated. Relative cumulus expansion and GSH levels were higher in the LFF group compared with in the MFF group after the IVM-I stage (P < 0.05). After the IVM-II stage, the numbers of oocytes in metaphase-II were increased in the LFF group and GSH content was higher in all of the LFF treatment groups compared with in the MFF treatment groups during both IVM stages (P < 0.05). ROS levels were reduced by LFF treatment regardless of IVM stage (P < 0.05). Blastocyst formation and the total numbers of cells in blastocysts were increased in all LFF treatment groups compared with the control group (P < 0.05). These results suggested that pFF from large follicles at the IVM stage could improve nucleic and cytoplasmic maturation status and further embryonic development through reducing ROS levels and enhancing responsiveness to gonadotropins.

296 IMPACT OF CO-CULTURING CUMULUS-ENCLOSED PORCINE OOCYTES WITH DENUDED OOCYTES DURING IN VITRO MATURATION IN A DEFINED MEDIUM ON CUMULUS EXPANSION AND OOCYTE MATURATION

2015 ◽  
Vol 27 (1) ◽  
pp. 237
Author(s):  
R. Appeltant ◽  
T. Somfai ◽  
M. Nakai ◽  
S. Bodo ◽  
D. Maes ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Binary Logistic Regression ◽  
Defined Medium ◽  
Positive Influence ◽  
Developmental Competence ◽  
Cumulus Expansion ◽  
Nuclear Maturation ◽  
Morphological Criteria

Recent research has revealed that oocyte-secreted factors (OSF) affect cumulus expansion and play important roles during maturation and embryo development of mammalian oocytes. The use of denuded oocytes (DO) as supplements during in vitro maturation (IVM) in a nondefined medium improved developmental competence of cumulus-enclosed porcine oocytes (COC; Gomez et al. 2012 Zygote 20, 135–145). We investigated the effect of DO on cumulus expansion and nuclear maturation of COC in pigs during IVM using a defined medium. If the DO exert a positive influence on IVM, the defined medium can then be analysed for the presence of OSF. Immature COC were collected in the slaughterhouse from prepubertal gilts. To obtain DO, some COC were completely denuded by pipetting through a narrow-bore glass pipette. The COC used as a source for DO fulfilled the same morphological criteria as the COC used for IVM. The IVM medium was porcine oocyte medium (POM; Yoshioka et al. 2008 J. Reprod. Dev. 54, 208–213) with hormone supplementations applied only during the first 20 h of the IVM period. The COC were fixed to the bottom of 35-mm plastic Petri dishes in 3 × 3 grids by Cell-Tak (BD Bioscience, Bedford, MA, USA) in 100-µL droplets POM covered by paraffin oil. Culture droplets (each including 1 COC grid) were supplemented with (DO+ group, n = 179) or without 16 DO (DO– group, n = 143). After 20 h of IVM, the medium was replaced with a preincubated hormone-free POM and oocytes were cultured for an additional 28 h. At 0, 20, and 48 h of IVM, images of each grid were taken at the same magnification. The size of each COC was measured as a 2-dimensional area in pixels by analysing images with ImageJ software. Relative cumulus expansion was calculated at 20 and 48 h of IVM on the basis of the initial COC size at 0 h, which was assigned as 1. At 48 h of IVM, the COC were denuded and examined for oocyte maturation by orcein staining. The experiment was replicated 5 times. Cumulus expansion ratios at 20 and 48 h of IVM were compared between the DO+ and DO– groups by ANOVA. Maturation rates were compared between the DO+ and DO– groups by binary logistic regression. No difference in cumulus expansion between DO– and DO+ could be observed at 20 h (1.83 ± 0.04 and 1.75 ± 0.03, respectively) and 48 h (1.41 ± 0.03 and 1.47 ± 0.02, respectively) of IVM. Nuclear maturation rates of COC in DO– and DO+ groups did not differ significantly (39.0 ± 5.4 and 32.9 ± 8.8%, respectively). In conclusion, addition of DO to the defined IVM medium did not affect the cumulus expansion and oocyte maturation of follicular porcine COC. Further research is needed to assess the effects of DO during IVM on subsequent fertilization. If DO prove to be beneficial for fertilization, the nature of the OSF will be investigated.This study was supported by FCWO of UGent and by FWO-Flanders (grant number FWO11/ASP/276).

355 DEVELOPMENTAL COMPETENCE OF IMMATURE BUBALINE CUMULUS - OOCYTE COMPLEXES VITRIFIED BY THE OPEN PULLED STRAW AND CONVENTIONAL STRAW METHODS

2007 ◽  
Vol 19 (1) ◽  
pp. 293
Author(s):  
A. Sharma ◽  
G. N. Purohit
Keyword(s):  
Morphological Changes ◽  
Developmental Competence ◽  
Control Group ◽  
Equal Concentration ◽  
Nuclear Maturation ◽  
Developmental Capacity ◽  
Phosphate Buffered Saline ◽  
Dose Dependent Increase ◽  
Dose Dependent

The in vitro maturation (IVM), fertilization (IVF), and morphological changes in buffalo cumulus–oocyte complexes (COCs) cryopreserved by ultrarapid freezing using conventional (CON) and open pulled staw (OPS) methods were tested. COCs were cryopreserved using a vitrification solution comprised of Dulbecco&apos;s phosphate-buffered saline+0.5 M sucrose+0.4% BSA and two concentrations (4.5 or 5.5 M) of each cryoprotectant ethylene glycol (EG) and dimethylsulfoxide (DMSO) by either the CON or the OPS method. Vitrified COCs were stored in LN for 7 days and then thawed; morphologically normal COCs were used for IVM (n = 1070) and IVF (n = 933) in 2 separate experiments to record morphological damage of COCs due to vitrification, nuclear maturation 24 h after culture (9 replicates), and fertilization 24 h after insemination (10 replicates). The COCs were matured in vitro in TCM-199 media with hormone supplements and fertilized using TALP-BSA as described previously (Purohit et al. 2005 Anim. Reprod. Sci. 87, 229–239). Freshly collected COCs were separately used for IVM (n = 110) and IVF (n = 130) and kept as controls. The arcsin transformed data of the proportions of oocytes matured or fertilized was compared by Duncan&apos;s new multiple range test. The highest proportion of morphologically normal oocytes was seen in 5.5 M EG with the CON method (94.5%) and the lowest was seen in 4.5 M DMSO with the OPS method (82.4%). At the end of experiment 1, it was apparent that IVM in all vitrification groups was significantly lower (P &lt; 0.05) compared to the control group (66.4%). Among the various vitrification treatments, the highest IVM occurred in 5.5 M EG with the OPS method (39.2%) and the lowest in 4.5 DMSO with the CON method (19.3%). Comparison of both concentrations of EG and DMSO showed that the proportion of COCs attaining Metaphase-II (M-II) increased with increasing concentration of both of the cryoprotectants. However, at equal concentration of EG and DMSO, the proportion of COCs attaining M-II was significantly higher in the OPS method compared to the CON method. In experiment 2, a significantly higher (P &lt; 0.05) IVF was seen for fresh COCs (45.4%) compared to vitrified COCs. Among the vitrification treatments, the highest fertilization was seen in 5.5 M EG with the OPS method (33.6%) and the lowest in 4.5 M DMSO with the CON method (15.17%). A dose-dependent increase in the proportion of oocytes fertilized was seen with increasing concentration of both EG and DMSO [CON: 4.5 M (15.2%), 5.5 M (25.6%); OPS: 4.5 M (21.3%), 5.5 M (27.5%)] in both CON and OPS methods. Comparison of the 2 cryoprotectants revealed that EG was better compared to DMSO.At equal concentrations of EG or DMSO, a significantly higher (P &lt; 0.05) proportion of fertilized oocytes was seen in the OPS method compared to the CON method. It was concluded that vitrification results in some damage to oocytes, with decrease in their subsequent IVM and IVF. Developmental capacity of vitrified buffalo oocytes can be improved by using OPS instead of conventional straws.

66 Bovine corpus luteum affects embryo developmental competence and production

2019 ◽  
Vol 31 (1) ◽  
pp. 158
Author(s):  
J. M. Alvarado ◽  
M. A. Tenemaza ◽  
S. L. Merchán ◽  
D. E. Argudo ◽  
M. S. Méndez ◽  
...  
Keyword(s):  
Corpus Luteum ◽  
Developmental Competence ◽  
Average Weight ◽  
Embryo Production ◽  
Fluid Medium ◽  
Bovine Corpus Luteum ◽  
Developmental Capacity ◽  
Recorded Data ◽  
Fetal Bovine

This study was conducted with the aim to determine the effect of the bovine corpus luteum on in vitro embryo production. Immature cumulus-oocyte complexes (COC) were aspirated from abattoir ovaries from cows (mainly Holstein and dairy crossbred cows and heifers) with (ipsilateral; CL+) and without (contralateral; CL−) corpus luteum (CL), and from cows without CL in any of the ovaries. The average weight of the ovaries from each group was 10.4±0.25, 5.7±0.25, and 6.1±0.25g for CL+, CL−, and C, respectively. The experiment was completed within 12 replicates (100 ovaries per group). The COC were cultured in maturation medium (TCM-199 supplemented with 10% fetal bovine serum, 100µg mL−1 sodium pyruvate, 0.75mg mL−1 of L-glutamine, 4µg mL−1 of FSH-p, 100 µM cysteamine, and 250µg mL−1 of gentamicin) followed by IVF (synthetic oviducal fluid medium supplemented with 10µg mL−1 heparin) and in vitro culture (citrate synthetic oviducal fluid medium). On Day 7 after IVF, the embryos were evaluated and classified into morulae (M), early blastocysts (EB), regular blastocysts (RB), expanded blastocysts (ExB), and hatched blastocysts (HB), and the embryos with quality 1 (according to IETS criteria) were recorded. Data were analysed by logistic regression and general linear model of SAS (SAS Institute Inc., Cary, NC, USA), and means were compared by the least squares means method. Results of cleavage, embryo rate at Day 7, and rates of M+EB, RB, ExB, and HB are shown in Table 1. The number of embryos per ovary was greater (P&lt;0.01) in CL+ (1.16±0.11) than in CL− (0.45±0.15) and C (0.55±0.15). Also, the number of embryos per cultured oocyte was significantly greater in CL+ than in CL− and C (0.27±0.02v. 0.14±0.03 and 0.15±0.03, respectively; P&lt;0.01). The results of this study reveal that the presence of the corpus luteum in the ovary at the time of the oocyte recovery affects the developmental capacity of the bovine embryos, and such influence probably occur through intraovarian interactions. Table 1.Effect of bovine corpus luteum on embryo production

scholarly journals Fluctuations in bovine ovarian follicular fluid composition throughout the oestrous cycle

Reproduction ◽  
2005 ◽  
Vol 129 (2) ◽  
pp. 219-228 ◽  
Author(s):  
Nicolas M Orsi ◽  
Nadia Gopichandran ◽  
Henry J Leese ◽  
Helen M Picton ◽  
Sarah E Harris
Keyword(s):  
Amino Acid ◽  
Follicular Fluid ◽  
Amino Acid Profile ◽  
Oestrous Cycle ◽  
Developmental Competence ◽  
Fluid Composition ◽  
Developmental Potential ◽  
Maturation Medium ◽  
Nutrient Profiles

Bovine oocyte maturation in vitro frequently results in abnormal cytoplasmic maturation and failure to acquire developmental competence. This is, in part, likely to be due to the non-physiological nutritional milieu to which oocytes are exposed. Improvements in oocyte developmental potential may be achieved by modelling nutrient profiles on those of preovulatory follicular fluid (FF). However, little is known about fluctuations in FF nutrient levels according to follicle dominance and oestrous cyclicity. This study therefore characterised the carbohydrate and amino acid profile of FF according to these parameters, and compared preovulatory FF composition with that of maturation medium. Carbohydrate concentrations (n = 121) were determined enzymatically whilst amino acid profiles (n = 40) were determined by reverse-phase HPLC. Pyruvate and glucose concentrations were unaffected by follicle dominance, whereas Stage III–IV lactate profiles were higher in non-dominant FF (P < 0.01). While most dominant FF amino acid concentrations were affected by oestrous stage, only glutamate, alanine, leucine and lysine levels fluctuated in non-dominant FF. Glucose and lactate concentrations were significantly negatively correlated, whereas most amino acids were significantly positively correlated with each other. Maturation medium had higher pyruvate and lower lactate concentrations than preovulatory FF (P < 0.001), whereas glucose level was similar. All amino acid levels (except histidine, taurine, alanine and tryptophan) differed significantly between maturation medium and preovulatory FF. These data indicated that FF composition varies throughout the oestrous cycle. Preovulatory FF nutrient profile differed from that of maturation medium, perhaps accounting for the poor developmental competence of in vitro matured oocytes. These data may contribute to the formulation of a nutritionally more physiological maturation medium.

scholarly journals Histone deacetylase inhibitor during in vitro maturation decreases developmental capacity of bovine oocytes

PLoS ONE ◽  
2021 ◽  
Vol 16 (3) ◽  
pp. e0247518
Author(s):  
Thais Preisser Pontelo ◽  
Mauricio Machaim Franco ◽  
Taynan Stonoga Kawamoto ◽  
Felippe Manoel Costa Caixeta ◽  
Ligiane de Oliveira Leme ◽  
...  
Keyword(s):  
Embryo Development ◽  
In Vitro Maturation ◽  
Embryo Quality ◽  
Cumulus Cells ◽  
Developmental Competence ◽  
Nuclear Maturation ◽  
Deacetylase Inhibitor ◽  
Developmental Capacity ◽  
Bovine Oocytes

This study aimed to evaluate the effect of scriptaid during pre-maturation (PIVM) and/or maturation (IVM) on developmental competence of bovine oocytes. Cumulus-oocyte complexes (COCs) were submitted to PIVM for 6 h in the presence or absence of scriptaid. COCs were distributed into five groups: T1-IVM for 22 h, T2-PIVM for 6 h and IVM for 22 h, T3-PIVM with scriptaid for 6 h and IVM for 22 h, T4-PIVM for 6 h and IVM with scriptaid for 22 h, and T5-PIVM with scriptaid for 6 h and IVM with scriptaid for 22 h. Nuclear maturation, gene expression, cumulus cells (CCs) expansion, and embryo development and quality were evaluated. At the end of maturation, all groups presented the majority of oocytes in MII (P>0.05). Only HAT1 gene was differentially expressed (P<0.01) in oocytes with different treatments. Regarding embryo development at D7, T4 (23%) and T5 (18%) had lower blastocyst rate (P<0.05) than the other treatments (T1 = 35%, T2 = 37% and T3 = 32%). No effect was observed when scriptaid in PIVM was used in less competent oocytes (P>0.05). In conclusion, presence of scriptaid in PIVM and/or IVM did not improve developmental competence or embryo quality.

scholarly journals Meiotic inhibition of bovine oocytes in medium supplemented with a serum replacer and hormones: effects on meiosis progression and developmental capacity

Zygote ◽  
2010 ◽  
Vol 19 (2) ◽  
pp. 107-116 ◽  
Author(s):  
Letícia Siqueira Sá Barretto ◽  
Viviane Sgobbi Dias Caiado Castro ◽  
Joaquim Mansano Garcia ◽  
Gisele Zoccal Mingoti
Keyword(s):  
Embryonic Development ◽  
Embryo Development ◽  
Developmental Competence ◽  
Meiotic Arrest ◽  
Nuclear Maturation ◽  
Developmental Capacity ◽  
Bovine Oocytes ◽  
Kinetics Of ◽  
Hatching Rates

SummaryAiming to improve the developmental competence of bovine oocytes during meiotic block, this study evaluated the effects of a serum replacer (Knockout SR®) and hormones (gonadotropins and estradiol) supplementation of prematuration medium (TCM119 with 0.5 mM IBMX [IBMX group] or 25 μM roscovitine [ROSC group]) on the kinetics of oocyte nuclear maturation and embryo development. Most IBMX and ROSC oocytes prematured for 8 h culture remained in the GV stage (70.3% and 73.1%, respectively; p > 0.05) similar to Control 8 h (63.5%) and to control immature oocytes (Control 0 h, 92.5%). After prematuration for 16 h, no oocytes remained in the GV stage at similar rates to those recently aspirated (p < 0.05); GV rates in ROSC (32.4%) were higher (p < 0.05) than in the Control 16 h group (8.6%), but similar (p > 0.05) to IBMX (9.7%). After in vitro maturation (IMV) for 24 h, metaphase II (MII) rates for oocytes prematured during 8 h were similar (p > 0.05) between control and treatments (65.0–71.7%). Similarly, MII rates oocytes prematured during 16 h were similar (p > 0.05) between all groups (45.9–60.4%). Cleavage rates (67.8–78.2%), embryonic development in day-7 (25.0–35.6%) and hatching rates in day-8 (2.5–11.3%) oocytes blocked during 8 h were similar for all groups (p > 0.05). Results indicate that addition of Knockout SR® and hormones to meiotic block culture with IBMX and roscovitine negatively affected meiotic arrest, but did not impair oocyte nuclear maturation and acquisition of developmental competence.

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