257 DEVELOPMENT OF HEAT-STRESSED OVA MATURED IN THE PRESENCE OF A PROSTAGLANDIN F2ALPHA RECEPTOR ANTAGONIST

2009 ◽  
Vol 21 (1) ◽  
pp. 226
Author(s):  
A. M. Ward ◽  
F. N. Schrick ◽  
R. R. Payton ◽  
E. Peixoto ◽  
J. L. Edwards
Keyword(s):  
Heat Stress ◽  
Receptor Antagonist ◽  
Fixed Effects ◽  
In Vitro Maturation ◽  
Elevated Temperatures ◽  
Block Design ◽  
Developmental Competence ◽  
In Vitro Production ◽  
Blastocyst Development

Studies in the literature have shown that cumulus–oocyte complexes produce PGF2α, that ova and cumulus cells have PGF2α receptors, and that PGF2α addition to maturing ova, above what would normally be produced, decreases blastocyst development. Because previous studies have shown elevated systemic and tissue levels of PGF2α as a consequence of heat stress, it was hypothesized that detrimental effects of exposing maturing ova to elevated temperatures may be mediated in part through heat-induced increases in PGF2α. To test this hypothesis, cumulus–oocyte complexes were matured at 38.5 or 41.0°C in the presence of a PGF2α receptor antagonist (AL-8810). Preattachment embryo development of AL-8810-treated ova was compared with development of ova matured in media with or without diluent (DMSO added at the same concentration as AL-8810; diluent and developmental controls, respectively), resulting in 6 total treatment combinations. Data were analyzed as a randomized block design (blocking on oocyte collection date) with fixed effects of maturation temperature, AL-8810, and the respective interaction included in the statistical model. In experimental replicates in which the effects of heat stress decreased blastocyst development greater than 10% (n = 14), a significant maturation temperature × AL-8810 interaction was noted when evaluating blastocyst development (P = 0.05). Specifically, when ova were heat stressed during the first 12 h of in vitro maturation, blastocyst development was reduced in developmental and diluent controls (26.2 v. 18.8 and 24.4 v. 19.9, respectively; SEM = 1.6). In contrast, when ova were matured under the same conditions but in the presence of a PGF2α receptor antagonist, the effects of heat stress to reduce blastocyst development after in vitro fertilization were no longer observed (22.5 v. 22.5; SEM = 1.6). When using abattoir-derived ovaries, it is not uncommon to collect, on occasion, ova that are developmentally challenged (i.e. blastocyst development is less than the 20 to 50% expected). In this experiment, this occurred on 5 occasions. Data from these experimental replicates were analyzed and reported separately because previous efforts had shown that the responsiveness of ova to changes in culture environment differs depending on the level of developmental competence. Relevant to this study, addition of AL-8810 to developmentally challenged ova matured under thermoneutral conditions increased cleavage (60.4 v. 55.4%, respectively; P = 0.06) and blastocyst development (17.7 v. 13.7%, respectively; P = 0.07). In summary, data illustrate that developmentally challenged ova, heat-stressed or otherwise, are susceptible to detrimental effects of PGF2α. The ability to increase blastocyst development approaching or exceeding the values expected for competent ova suggests the usefulness of a PGF2α receptor antagonist during in vitro maturation to improve the efficiency of in vitro production procedures.

Effect of antioxidant ascorbic acid on in vitro maturation of Caprine oocytes under normal and elevated temperatures

2018 ◽  
Author(s):  
S.B. Khanday ◽  
J.A. Ahmed ◽  
N. Nashiruddullah ◽  
U. Sharma and D. Chakraborty
Keyword(s):  
Ascorbic Acid ◽  
Heat Stress ◽  
In Vitro Maturation ◽  
Elevated Temperatures ◽  
Developmental Competence ◽  
Cumulus Expansion ◽  
Nuclear Maturation ◽  
Maturation Rate ◽  
Metaphase Stage

The aim of the present study was to assess the effect of ascorbic acid on in vitro maturation of caprine oocytes under normal and elevated temperatures. Goat ovaries were collected at slaughter and both A and B grade cumulus-oocyte-complexes (COCs) were aspirated out and were matured in vitro under normal (38.5°C) and elevated temperatures (41°C). On the basis of cumulus expansion and nuclear maturation, the maturation competencewere compared with and without ascorbic acid supplementation (100 µM). Heat stress significantly (P£ 0.01) reduced cumulus expansion, maturation rate and lowered metaphase stage II of nuclear maturation. Ascorbic acid improved developmental competence of oocytes during heat stress (41 °C) and ascorbic acid supplemented COCs demonstrated significantly (P£ 0.05) higher maturation rates when compared to non-supplemented groups.

233. Oxygen concentration during in vitro maturation of murine oocytes affects blastocyst cell lineage

2005 ◽  
Vol 17 (9) ◽  
pp. 91
Author(s):  
K. M. Banwell ◽  
M. Lane ◽  
D. L. Russell ◽  
K. L. Kind ◽  
J. G. Thompson
Keyword(s):  
Embryo Development ◽  
In Vitro Maturation ◽  
Cell Lineage ◽  
Developmental Competence ◽  
Embryo Viability ◽  
Blastocyst Development ◽  
Treatment Groups ◽  
Significant Difference ◽  
O2 Concentration

Follicular antral oxygen tension is thought to influence subsequent oocyte developmental competence. Despite this, in vitro maturation (IVM) is routinely performed in either 5 or 20% O2 and while low O2 has been shown to be beneficial to embryo development in many species, the effect of altering O2 concentration during IVM has not been adequately investigated. Here we investigated the effects of a range of O2 concentrations during IVM on meiotic maturation and subsequent embryo development after IVF. Ovaries from eCG-stimulated CBA F1 female mice (21 days) were collected and intact cumulus oocyte complexes (COCs) cultured for 17–18 h under 2, 5, 10 or 20% O2 (6% CO2 and balance of N2). Matured COCs were denuded of cumulus cells, fixed and stained (1% aceto-orcein) for visualisation of maturation status. No significant difference in maturation rates between treatment groups was observed. Following IVF (performed under 5% O2, 6% CO2 and balance of N2), no difference in fertilisation rates between treatment groups was observed in a randomly selected cohort 7 h post-fertilisation. There was also no significant difference in cleavage rates after 24 h or ability to reach blastocyst stage after 96 h, with a tendency (P = 0.079) for more blastocysts in 2% O2. However there was a significant increase in the number of trophectoderm cells present in the resulting blastocysts (P < 0.05) in the 2% O2 group (35 ± 2.1) compared to 20% O2 (25 ± 2.8). Our data suggests that O2 concentration during IVM does not influence nuclear maturation or subsequent fertilisation, cleavage and blastocyst development rates. However, maturation in 2% O2 significantly alters subsequent cell lineage within blastocysts to favour trophectoderm development. Such skewed trophectoderm cell number may influence embryo viability. Funded by NHMRC and NIH.

Relationship between follicle size and oocyte developmental competence in prepubertal and adult pigs

2007 ◽  
Vol 19 (7) ◽  
pp. 797 ◽  
Author(s):  
Melanie A. Bagg ◽  
Mark B. Nottle ◽  
David T. Armstrong ◽  
Christopher G. Grupen
Keyword(s):  
In Vitro Maturation ◽  
Fold Increase ◽  
Cell Number ◽  
Developmental Competence ◽  
Blastocyst Development ◽  
Parthenogenetic Activation ◽  
Follicle Size ◽  
Low Efficiency ◽  
The Relationship

The present study compared the distribution and steroid composition of 3-, 4- and 5–8-mm follicles on the surface of prepubertal and adult ovaries, and determined the relationship between follicle size and developmental competence of oocytes following parthenogenetic activation. The effect of 1 mm dibutyryl cAMP (dbcAMP) for the first 22 h of in vitro maturation (IVM) on the embryo development of prepubertal oocytes from the three follicle size cohorts was also determined. Compared with adult, prepubertal ovaries contained a higher proportion of 3-mm follicles (46 v. 72%, respectively), but a lower proportion of 4-mm (33 v. 22%, respectively) and 5–8-mm follicles (21 v. 6%, respectively). Adult follicular fluid (FF) contained 11-fold higher levels of progesterone (P4) than prepubertal FF, with similar levels observed between all adult follicle sizes. In prepubertal FF, the P4 concentration increased with follicle size from 3 to 4 to 5–8 mm. Rates of blastocyst development following parthenogenetic activation of adult oocytes from all three follicles sizes were similar (approximately 55%), whereas rates from prepubertal oocytes increased with increasing follicle size from 3 (17%) to 4 (36%) to 5–8 mm (55%). Treatment with dbcAMP for the first 22 h of IVM led to a 1.5-fold increase in the rate of blastocyst development for prepubertal oocytes from 3-mm follicles, but had no effect on prepubertal oocytes from the 4 and 5–8 mm classes. Mean blastocyst cell number increased with follicle size in prepubertal ovaries and was similar for all follicle sizes in adult ovaries. The present study demonstrates that the low efficiency of in vitro embryo production observed using prepubertal compared with adult pig oocytes is due to a greater proportion of 3-mm follicles on prepubertal ovaries, which contain oocytes of inferior developmental competence.

scholarly journals Effects of in vitro maturation media on in vitro fertility of porcine oocytes and early development of embryos

2020 ◽  
Vol 18 (2) ◽  
pp. 249-255
Author(s):  
Nguyen Viet Linh ◽  
Nguyen Thi Hiep
Keyword(s):  
Follicular Fluid ◽  
In Vitro Maturation ◽  
Subsequent Development ◽  
Developmental Competence ◽  
In Vitro Production ◽  
Vitro Fertilization ◽  
Fetal Bovine ◽  
Vitro Production ◽  
Normal Fertilization

In pigs, embryo productivity is still lower than that in other livestocks. One of the reasons is incomplete maturation of porcine oocytes in in vitro conditions. Therefore in vitro maturation (IVM) plays a crucial role in in vitro production of porcine embryos. It provides prerequisite condition to in fertilization and subsequent development of porcine embryos. In a previous study, effects of NCSU-37-based medium and TCM-199-based media supplemented with porcine follicular fluid (pFF) or Fetal Bovine Serum (FBS) on in vitro maturation of Landrace oocytes collected in Vietnam have been compared, suggesting that NCSU-37 medium supplemented with 10% of porcine follicular fluid (pFF) had the highest rate of oocytes reach to metaphase II stage in comparison to those of the other two TCM-199-based media. In the present study, further experiments were carried out to evaluate the contribution of IVM media on fertilization capability and developmental competence. Porcine oocytes matured in vitro in 3 media: NCSU-37 supplemented with 10% pFF, TCM-199 supplemented with either 10% pFF or 10% FBS were subjected to in vitro fertilization and subsequent in vitro culture to monitor fertility and embryo development. The results showed that penetration and normal fertilization rates in both TCM-199 groups are both higher than that of NCSU-37 group. Moreover, the cleavage and blastocyst rates, and cell numbers of blastocysts which is a criterion for embryo quality were all higher in TCM-199 groups, especially in the group supplemented with pFF. It might be concluded that TCM-199 media supplemented with either pFF or FBS are suitable for effective in vitro maturation of Landrace porcine oocytes collected in Vietnam.

scholarly journals Effect of heat stress during in vitro maturation on developmental competence of vitrified bovine oocytes

2017 ◽  
Vol 52 ◽  
pp. 48-51 ◽  
Author(s):  
M Vendrell-Flotats ◽  
N Arcarons ◽  
E Barau ◽  
M López-Béjar ◽  
T Mogas
Keyword(s):  
Heat Stress ◽  
In Vitro Maturation ◽  
Developmental Competence ◽  
Bovine Oocytes

scholarly journals Effect of varying glucose and glucosamine concentration in vitro on mouse oocyte maturation and developmental competence

2013 ◽  
Vol 25 (8) ◽  
pp. 1095 ◽  
Author(s):  
L. A. Frank ◽  
M. L. Sutton-McDowall ◽  
D. L. Russell ◽  
X. Wang ◽  
D. K. Feil ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Blastocyst Stage ◽  
Developmental Competence ◽  
Blastocyst Development ◽  
Oocyte Competence ◽  
Hexosamine Pathway ◽  
Hexosamine Biosynthesis ◽  
Necessary And Sufficient

The effects of hyper- and hypo-glycaemic conditions during the in vitro maturation of mouse cumulus–oocyte complexes on developmental competence were examined, with an emphasis on the role of the hexosamine biosynthesis pathway. A low (1 mM) glucose concentration achieved optimal oocyte competence (3-fold higher blastocyst development rate compared with high (30 mM) glucose, P < 0.05). In addition, glucose supplementation during only the first hour after release from the follicle was necessary and sufficient to support oocyte maturation and embryo development to the blastocyst stage. Glucosamine (a known hyperglycaemic mimetic and specific activator of the hexosamine pathway) was able to substitute for glucose during this first hour, indicating that flux through the hexosamine pathway is essential for oocyte competence. In the absence of glucose throughout the maturation period, glucosamine was not able to increase developmental competence, and at higher concentrations (2.5 and 5 mM) had a detrimental effect on MII and blastocyst development rates, compared with controls (P < 0.05). These experiments underscore the importance of glucose metabolic pathways during in vitro maturation and support the concept that excess flux through the hexosamine pathway has detrimental consequences.

196 ELEVATED TEMPERATURE DURING BOVINE OOCYTE MATURATION ALTERS MITOCHONDRIAL ACTIVITY

2016 ◽  
Vol 28 (2) ◽  
pp. 229
Author(s):  
R. R. Payton ◽  
L. A. Rispoli ◽  
K. A. Nagle ◽  
A. M. Saxton ◽  
J. L. Edwards
Keyword(s):  
Heat Stress ◽  
Fixed Effects ◽  
Block Design ◽  
Blastocyst Stage ◽  
Mitochondrial Activity ◽  
Atp Content ◽  
Total Glutathione ◽  
Acetoxymethyl Ester ◽  
Mitochondrial Ros

Exposure of maturing cumulus-oocyte complexes to a physiologically relevant heat stress altered the transcriptome in oocytes, especially certain transcripts important for mitochondrial function. To determine if perturbations are coincident with changes in mitochondrial activity, generation of reactive oxygen species (ROS), glutathione and ATP content were examined in maturing oocytes experiencing heat stress. In the first study, ROS content was assessed after 6 h at 38.5, 41, or 42°C using 37 µM 6-carboxy-2′7′-dichlorodihydrofluorescein diacetate, di(acetoxymethyl ester) or 60 µM dihydrofluorescein diacetate for staining cytoplasmic and mitochondrial ROS, respectively. In a second study, cumulus-oocyte complexes were matured for 0, 12 or 24 h at 38.5°C (control) or 41°C for 12 h (heat stress; 38.5°C thereafter). Total glutathione (GSSG + GSH) and reduced glutathione (GSH) and ATP levels were assessed in cumulus denuded-zona free oocytes as per manufacturer (GSH-Glo Glutathione assay, Promega, Madison, WI, USA; 1 to 25 pmol standards) and ATP determination kit (Life Technologies, Carlsbad, CA, USA; 0.078 to 10 pmol standards). Next, ATP content was assessed in control and heat-stressed oocytes matured for 24 h and in embryos resulting after IVF of control or heat-stressed oocytes (i.e. 2-, 4-, and 8-to-16 cell and blastocyst-stage embryos). Data were analysed as a randomised block design with fixed effects of maturation temperature (and hours of maturation where appropriate), blocking on replicate, using PROC MIXED (SAS 9.2, SAS Institute, Cary, NC, USA). Culture at 41°C for the first 6 h in vitro-matured (IVM) reduced levels of cytoplasmic ROS compared to non-stressed controls (1.02 v. 0.84 fluorescent ratio for 38.5 v. 41°C, SEM = 0.16, P < 0.0001), whereas levels after 42°C were similar to controls (0.94). Mitochondrial ROS were higher after 41°C (1.42) than after 38.5 (0.98) or 42°C (0.98, SEM = 0.14, P < 0.0001). Heat stress exposure increased total glutathione content at 12 h (4.4 v. 5.2 pmol for 38.5 v. 41°C) but levels were decreased by 24 h (6.6 v. 5.9 pmol for 38.5 v. 41°C; SEM = 1.26, P = 0.0002). A similar pattern was found for GSH (P = 0.0001); GSSG was similar across treatments (P = 0.50). Levels of ATP increased during maturation (1.10, 1.20, and 1.65 pmol for 38.5°C at 0, 12, and 24 h) and were increased by heat stress at 24 h (1.65 and 2.01 pmol for 38.5 and 41°C; SEM = 0.31, P = 0.001). In embryos, ATP content was higher in 8-to-16 cell embryos derived from heat-stressed oocytes than in those from control oocytes (P = 0.03); levels were similar in blastocyst-stage embryos (P = 0.21) regardless of origin. In conclusion, application of a physiologically relevant heat stress during maturation altered mitochondrial activity in bovine oocytes. Carryover of effects to the early embryo may explain some of the reductions in embryo development experienced by heat-stressed oocytes. This research was supported in part by USDA National Institute of Food and Agriculture, Hatch Project No. 227701, the state of Tennessee through University of Tennessee AgResearch, Department of Animal Science, and East Tennessee Research and Education Center.

163 EFFECTS OF DIFFERENT IN VITRO MATURATION SYSTEMS ON EMBRYO DEVELOPMENT IN BOVINE PREPUBERTAL AND ADULT DONORS

2014 ◽  
Vol 26 (1) ◽  
pp. 195
Author(s):  
S. M. Bernal ◽  
J. Heinzmann ◽  
D. Herrmann ◽  
U. Baulain ◽  
A. Lucas-Hahn ◽  
...  
Keyword(s):  
Gene Expression ◽  
Embryo Development ◽  
In Vitro Maturation ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Developmental Competence ◽  
In Vitro Production ◽  
Lactating Cows

Prepubertal bovine females have been suggested as a source of oocytes in order to accelerate genetic gain and decrease the generation interval. However, prepubertal oocytes have a lower developmental competence than their adult counterparts. In vitro maturation (IVM) systems using cyclic AMP (cAMP) regulators and 30-h culture have been suggested to improve blastocyst in vitro production rates from bovine oocytes (Albuz et al., 2010). The present study evaluated the effects of an addition of the cAMP modulators forskolin, 3-isobutyl-1-methylxanthine (IBMX), and cilostamide during extended IVM on blastocyst yields and gene expression in prepubertal and adult bovine females. Holstein-Friesian donors were submitted to ovum pick-up twice per week. Oocytes from groups of 12 animals, including lactating cows (>2 lactations) and prepubertal donors (6–10 months old) were used in the following treatment groups: TCM24 (24-h IVM, routine protocol/control), cAMP30 (2-h pre-IVM culture using forskolin-IBMX and 30-h IVM adding cilostamide), DMSO30 [2-h pre-IVM culture and 30-h IVM with dimethyl sulfoxide (DMSO)/vehicle control]. In vitro-matured oocytes were fertilized and presumptive zygotes were cultured in vitro to assess embryo development. In vivo blastocysts were produced from superovulated cows and used for gene expression analysis. Cleavage rates, blastocyst formation, and mRNA abundance of selected genes were evaluated. The Glimmix procedure from SAS/STAT (SAS Institute Inc., Cary, NC, USA) was performed to compare blastocyst and cleavage rates. One-way ANOVA was implemented to evaluate gene expression. A total of 793 oocytes from the different sources were submitted to the IVM treatments. Cleavage rates (prepubertal donors: 64.6 ± 4%, 59.1 ± 6.4%, 53 ± 4.4%, cows: 55.1 ± 4.3%, 59 ± 6.5%, 50.8 ± 4.4%, for TCM24, cAMP30, and DMSO30, respectively; P > 0.05) and blastocyst/zygotes rates (prepubertal donors: 27 ± 6%; 21.8 ± 3.5%; 17.6 ± 2.4%; cows: 28 ± 3.3%; 27.7 ± 2.9%; 22.7 ± 3.2% for TCM24, cAMP30, and DMSO30, respectively; P > 0.05) did not differ among in vitro treatments. The mRNA relative abundance of the EGR1 gene was down-regulated 6-fold in all in vitro-produced blastocysts compared with their in vivo counterparts (P < 0.05). Gene expression profiles for SLC2A8, DNMT3B, BCL-XL, and PRDX1 were similar in in vitro and in vivo blastocysts. These results show similar embryo production patterns in prepubertal and adult donors. Furthermore, DMSO did not show effects on embryo developmental rates when used during IVM. The gene expression levels of EGR1 confirm our recent findings in blastocysts obtained from oocytes from slaughterhouse ovaries (data not presented), showing its usefulness as an embryo quality marker. These preliminary results indicate that oocyte developmental capacity in prepubertal donors can be similar to that of the adult donors without addition of cAMP modulators.

scholarly journals 132 BLASTOCYST DEVELOPMENT OF EQUINE OOCYTES WITH LOW MEIOTIC COMPETENCE HELD IN ROSCOVITINE BEFORE IN VITRO MATURATION

2005 ◽  
Vol 17 (2) ◽  
pp. 216
Author(s):  
Y.H. Choi ◽  
L.B. Love ◽  
D.D. Varner ◽  
K. Hinrichs
Keyword(s):  
Epithelial Cells ◽  
In Vitro Maturation ◽  
Developmental Competence ◽  
Blastocyst Development ◽  
Exact Test ◽  
Nuclear Maturation ◽  
Significant Difference ◽  
Chromatin Configuration ◽  
Meiotic Competence

At the time of recovery, immature equine oocytes may be separated into those with either expanded cumuli (Ex) or compact cumuli (Cp). The Cp oocytes originate from viable follicles but are largely juvenile, with low meiotic competence (20 to 30% maturation to MII), and possibly reduced developmental competence. We previously found that in Cp oocytes recovered immediately after slaughter, suppression of meiosis with roscovitine for 24 h before maturation increased embryo development at 4 days after intracytoplasmic sperm injection (ICSI; Franz et al. 2003 Reproduction 125, 693–700). The present study was conducted to evaluate the effect of roscovitine suppression on nuclear maturation and blastocyst formation of Cp oocytes recovered after transport of ovaries from the abattoir (i.e. recovered 5–9 h after slaughter). Compact oocytes recovered from transported ovaries were cultured in M199 with 10% FBS containing 66 μM roscovitine with or without an oil cover. After 16–18 or 24 h, oocytes were fixed to examine the chromatin configuration. Treatment for 16–18 h without oil resulted in the lowest rate of meiotic resumption (0%); thus this treatment was utilized in further studies. Resumption in other treatments ranged from 3 to 6%. Following roscovitine suppression, oocytes were cultured for 30 h in M199 with 10% FBS and 5 μU mL−1 FSH for maturation; control oocytes were cultured for 30 h in the same medium immediately after recovery. Mature oocytes were subjected to ICSI, then cultured in DMEM/F-12 with 10% FBS with or without co-culture with equine oviductal epithelial cells under mineral oil in 5% CO2 in air at 38.2°C, and then evaluated at 7.5 days. Progression to MII (82/376, 22%) after maturation of roscovitine-treated oocytes was similar to that for control oocytes (74/395, 19%). There was no significant difference in cleavage rates after ICSI (72–78%) among treatments. Development to blastocyst was highest in roscovitine-treated oocytes in DMEM/F-12 with co-culture (11/30, 37%); this was significantly higher than that of non-treated oocytes in DMEM/F-12 alone (5/36, 14%), but similar to that of non-treated/DMEM/F-12/co-culture (10/37, 27%) and roscovitine/DMEM/F-12 alone (8/39, 21%). These data indicate that roscovitine induces a fully reversible meiotic suppression in Cp equine oocytes recovered 5–9 h after slaughter, and that this suppression does not harm subsequent developmental competence. This treatment may be used to manipulate the time of onset of maturation of equine oocytes for ease of subsequent procedures. Co-culture with oviductal epithelial cells tended to increase blastocyst rate (P = 0.1, Fisher's exact test) in contrast to our previous findings with embryos from Ex oocytes (Choi et al. 2004 Biol. Reprod. 70, 1231–1238). Further work is needed to determine whether this is related to differences in intrinsic developmental competence between oocyte types. This work was supported by the Link Equine Research Endowment Fund (Texas A&M University).

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