202 CHARACTERIZATION OF BOVINE OOCYTE CYTOPLASMIC MATURATION WITH COMMON IN VITRO MATURATION PROTOCOLS

2016 ◽  
Vol 28 (2) ◽  
pp. 232
Author(s):  
B. A. Foster ◽  
F. A. Diaz ◽  
P. T. Hardin ◽  
E. J. Gutierrez ◽  
K. R. Bondioli
Keyword(s):  
Gap Junctions ◽  
Gap Junction ◽  
In Vitro Maturation ◽  
Transvaginal Ultrasound ◽  
Bovine Oocyte ◽  
Hoechst 33342 ◽  
Nuclear Maturation ◽  
Mitochondrial Distribution ◽  
Cytoplasmic Maturation

Modulators of 3′-5′-cyclic adenosine monophosphate have been extensively researched to delay nuclear maturation in in vitro maturation (IVM) systems to improve synchronization of nuclear and cytoplasmic maturation. However while normal maturation for many organelles has been characterised, there is a lack of information on how modulators affect cytoplasmic maturation. The goal of this study was to identify the effect of different components of bovine oocyte maturation systems on 3 aspects of cytoplasmic maturation. Bovine oocytes were collected from mixed breed beef cattle using transvaginal ultrasound guided oocyte aspiration. Oocytes were assigned to 1 of 4 treatments; staining immediately after collection (n = 249) or after 24 h of IVM (n = 270), 2 h of pre-IVM in Forskolin and 3-isobutyl-1-methylxanthine (IBMX; n = 254), or 2 h of pre-IVM followed by IVM (n = 259). Following treatment, half of the recovered oocytes were stained with Hoechst 33342 to determine nuclear maturation status, and Calcein AM for gap junction status. The other half were stained with Hoechst 33342, Mitotracker deep red to identify mitochondria distribution patterns and Alexa Fluor 488 conjugated phalloidin for F actin microfilament distribution. Organelle patterns were coded and statistically analysed using linear models to determine if treatment had an effect on the indicators of cytoplasmic maturation or their agreement with nuclear maturation. Results indicated that there was a high degree of variability in both cytoplasmic and nuclear maturation of oocytes irrespective of treatment group, with many oocytes exhibiting aberrant patterns in both mitochondrial and microfilament distribution. Gap junctions were classified as open (immature), partially open or closed (mature), based on the strength of Calcein fluorescence within the ooplasm. Both treatment and nuclear maturation had a significant effect on gap junction status (P < 0.001) with gap junctions tending to close as oocytes matured, while treatment in pre-IVM maintained open gap junctions, even as meiosis progressed. Mitochondria were classified as peripheral (immature), diffuse, central (mature) or too sparse to accurately classify. There was an unexpectedly high proportion of oocytes with few mitochondria (17%), suggesting an incomplete growth phase before collection. There was no correlation between meiotic stage and mitochondrial distribution (P = 0.73), with the majority of oocytes having diffuse mitochondrial distribution. As normal maturation proceeds, microfilaments aggregate and migrate peripherally. However, neither microfilament aggregation nor redistribution were correlated with nuclear maturation (P = 0.6 and P = 0.11 respectively) or mitochondrial distribution (P = 0.33 and P = 0.06 respectively). Overall, results show that while pre-IVM maintains open gap junctions, the system studied here is not sufficient for improving correlation between cytoplasmic and nuclear maturation. Many deviations from normal cytoplasmic maturation are seen with IVM and these irregularities are maintained with prematuration in Forskolin and IBMX.

336 MEIOTIC DEVELOPMENT AND CORTICAL GRANULES DISTRIBUTION IN CANINE OOCYTES DURING IN VITRO MATURATION

2010 ◽  
Vol 22 (1) ◽  
pp. 324 ◽  
Author(s):  
M. De los Reyes ◽  
D. Luna ◽  
J. Palomino
Keyword(s):  
In Vitro Maturation ◽  
Germinal Vesicle ◽  
Fluorescein Isothiocyanate ◽  
Cumulus Cells ◽  
Homogeneous Distribution ◽  
Cortical Granules ◽  
Dapi Staining ◽  
Nuclear Maturation ◽  
Cytoplasmic Maturation

Low development of IVM canine oocytes could be in part attributed to an impaired cytoplasmic maturation. In mammalian oocytes, migration and the redistribution of cortical granules (CGs) around the periphery of the oocyte contribute to the inhibition of polyspermy and it is an important criterion to evaluate cytoplasmic maturation. The state of nuclear maturation and the distribution of CGs were evaluated in canine oocytes cultured for different periods in order to compare the synchrony of nuclear and cytoplasmic maturation during in vitro maturation. Bitch ovaries at different stages of the estrous cycle were obtained following ovariectomy. COCs with compact cumulus cells showing a homogeneous cytoplasm were selected for experiments. Thirty-six COCs were processed at immature stage, placed in PBS medium until evaluation. A total of 275 COCs were matured in vitro for 48, 72, and 96 h in TCM-199 with Earle’s salt supplemented with 25 mM Hepes, 10% FCS, 0.25 mM pyruvate, 10 IU mL-1 of hCG, 300 IU mL-1 penicillin, and 20 mg mL-1 streptomycin, at 38.5°C and 5% CO2. At each culture period, the oocytes were stained with Lens culinaris agglutinin (LCA), labeled with fluorescein isothiocyanate, and the CGs distributions were examined under a fluorescent microscope. The nuclear status of the denuded oocytes was determined by DAPI staining under a fluorescence microscope. For each treatment, at least four replicates were performed and the data was analyzed by ANOVA using Tukey’s test to determine the differences P < 0.05. Three types of CGs distribution were distinguished during canine oocyte maturation: (1) homogeneous distribution throughout the cytoplasm including the cortex; (2) heterogeneous (clusters) within the cytoplasm and (3) densely distributed beneath the oolemma. Nuclear stages were classified as immature or germinal vesicle (GV) stage; resumption of meiosis or germinal vesicle break down (GVBD); metaphase I to telophase I (MI toTel I); and mature or second metaphase (MII). The distribution patterns of GCs were different (P < 0.05) among oocytes cultured for different periods and the nuclear maturation status also differed between oocytes cultured for different intervals (P < 0.05). Most (>84%) of the immature oocytes at GV showed a uniform distribution of CGs throughout the cytoplasm. At 48 h of culture, CGs distribution was mainly Type 2 (25%) and 3 (61%) and the oocytes were at GVBD (33%) and MI-Tel I (33%) stages. Most nuclei of the type 3 oocytes were in the MI (40%) and MII (11%) stages, corresponding to those oocytes matured for 72 (88%) or 96 h (71%). These results indicate that canine oocytes migrate to the cortex during IVM and this process is not finished before 72 h of culture. In addition, although the re-distribution of the CGs occurred in parallel with nuclear maturation, the oocytes cannot always proceed to the MII stage; however, in such oocytes the CGs are distributed beneath the oolemma. Supported by Grant FONDECYT 1080618.

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.

scholarly journals Development of a reliable in vitro maturation system for zebrafish oocytes

Reproduction ◽  
2008 ◽  
Vol 135 (3) ◽  
pp. 285-292 ◽  
Author(s):  
Shinsuke Seki ◽  
Toshimitsu Kouya ◽  
Ryoma Tsuchiya ◽  
Delgado M Valdez ◽  
Bo Jin ◽  
...  
Keyword(s):  
In Vitro Maturation ◽  
Saline Solution ◽  
Stage Iii ◽  
Hatching Rate ◽  
Cleavage Rate ◽  
Immature Oocytes ◽  
Nuclear Maturation ◽  
Cytoplasmic Maturation

In zebrafish oocytes, it has been reported that a 60 or 75% Leibovitz L-15 medium or simple balanced saline solution containing 17α, 20β-dihydroxy-4-pregnen-3-one (DHP) is effective for nuclear maturation. However, most of the oocytes that matured under these conditions were not fertilized and did not hatch. Thus, thesein vitromaturation methods could not support the cytoplasmic maturation of zebrafish oocytes. Therefore, we tried to develop a reliablein vitromaturation method for zebrafish oocytes, which supports their ability to be fertilized and to develop till hatching. When zebrafish oocytes at stage III were cultured in 50–100% Leibovitz L-15 medium supplemented with DHP, the highest rates of cleavage (24%) and hatching (12%) were obtained from oocytes matured in 90% Leibovitz L-15 medium. When we examined the suitable pH (7.5–9.5) of the 90% medium, higher rates of cleavage (45%) and hatching (33%) were obtained in oocytes matured at pH 9.0 than at pH 7.5, 8.5, or 9.5 (cleavage rate, 16–29%; hatching rate, 8–21%). In oocytes matured in 90% Leibovitz L-15 medium at pH 9.0, high rates of cleavage (70%) and hatching (63%) were obtained when oocytes were cultured for 270 min with 0.5 mg/ml BSA. Thus, 90% Leibovitz L-15 medium at pH 9.0 containing 0.5 mg/ml BSA was effective for normal maturation of zebrafish oocytes. This method will become a powerful tool for understanding the mechanism ofin vitromaturation in zebrafish oocytes and for the practical use of immature oocytes.

scholarly journals Oxygen tension and oocyte density during in vitro maturation affect the in vitro fertilization of bovine oocytes

2015 ◽  
Vol 36 (6Supl2) ◽  
pp. 4277
Author(s):  
Angelo Bertani Giotto ◽  
Daniela Dos Santos Brum ◽  
Francielli Weber Santos ◽  
Antonio Carlos Galarça Guimarães ◽  
Cibele Garcia Moreira Gonçalves ◽  
...  
Keyword(s):  
Antioxidant Activity ◽  
In Vitro Fertilization ◽  
Oxygen Tension ◽  
In Vitro Maturation ◽  
Ros Production ◽  
Vitro Fertilization ◽  
Nuclear Maturation ◽  
Cytoplasmic Maturation ◽  
Bovine Oocytes

Oocyte maturation is the key factor affecting the fertilization and embryonic development. Factors such as oocyte density and oxygen tension can directly influence the IMV. Thus, the objective of this study was to evaluate the effect of the association of oxygen tensions (5% or 20%) with different oocyte densities (1:10?l or 1:20?l) in the in vitro maturation (IVM) of bovine oocytes on maturation and fertilization rates, ROS production and antioxidant activity. Three experiments were performed with bovine oocytes that were obtained from slaughterhouse ovaries. After selection, the oocytes were randomly distributed in four treatments: 1:10/5%; 1:10/20%; 1:20/5%and 1:20/20% for each experiment. In experiment I, nuclear maturation status and cytoplasmic maturation were evaluated through detection of the first polar body by immunofluorescence and the mitochondrial reorganization assay. In experiment II, ROS production and antioxidant activity were analyzed in oocytes and IVM medium after 24 h of maturation through detection of ROS, reduced glutathione (GSH) and Superoxide dismutase activity by spectrofluorimetric methods. In experiment III, fertilization was evaluated through pronucleus formation, sperm penetration with or without decondensation and polyspermy rates by immunofluorescence. In experiment I, the nuclear maturation and cytoplasmic maturation were similar among treatments (P>0.05). In experiment II, reactive oxygen species in oocytes were elevated in treatments with low oxygen tension which was independent of oocyte density (P<0.05). Additionally, ROS levels in IVM medium were higher in treatments with high oocyte density by volume of medium, which was independent of oxygen tension (P<0.05). In Experiment III, the fertilization and penetration rates were higher in the treatment with 20% oxygen tension and high oocyte density (P<0.05). Furthermore, a high incidence of polyspermy was observed in groups with high oxygen tension and low oocyte density (P<0.05). In conclusion, the results of this study indicate an interaction between oxygen tension and oocyte density, which increases ROS production in certain associations and subsequently influences the rates of in vitro fertilization of bovine oocytes. The improved rates of IVF were obtained when IVM was conducted using 20% oxygen tension and high oocyte density (1:20 ul).

Enhancement of lipid metabolism with L-carnitine during in vitro maturation improves nuclear maturation and cleavage ability of follicular porcine oocytes

2011 ◽  
Vol 23 (7) ◽  
pp. 912 ◽  
Author(s):  
Tamás Somfai ◽  
Masahiro Kaneda ◽  
Satoshi Akagi ◽  
Shinya Watanabe ◽  
Seiki Haraguchi ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
In Vitro Maturation ◽  
Blastocyst Stage ◽  
In Vitro Fertilisation ◽  
Mitochondrial Activity ◽  
Nuclear Maturation ◽  
Mitochondrial Functions ◽  
Cytoplasmic Maturation ◽  
Further Development

The aim of the present study was to assess the effects of L-carnitine, an enhancer of lipid metabolism and mitochondrial activity, during in vitro maturation (IVM) on nuclear maturation and in vitro fertilisation of porcine follicular oocytes and subsequent embryo development. Mitochondrial functions, intracellular lipid content and reactive oxygen species (ROS) levels in oocytes were also investigated. L-carnitine supplementation in 0.6–5 mg mL–1 concentration during IVM significantly improved (P < 0.05) the rates of metaphase-II (MII) stage oocytes compared with the control; however, fertilisation rates and monospermy were not improved. Although supplementation of IVM medium with L-carnitine significantly increased oocyte cleavage (P < 0.05), further development to the blastocyst stage was not improved. The density of active mitochondria was significantly higher and the density of lipid droplets was significantly lower (P < 0.05) in L-carnitine-treated oocytes compared with the control. Furthermore, the ROS levels in L-carnitine-treated oocytes were significantly lower than those in the control. In conclusion, enhancing mitochondrial functions by L-carnitine improved oocyte maturation and cleavage underlining the importance of lipid metabolism for nuclear and cytoplasmic maturation of porcine oocytes.

271 NUCLEAR MATURATION OF WOOD BISON (BISON BISON ATHABASCAE) CUMULUS - OOCYTE COMPLEXES

2013 ◽  
Vol 25 (1) ◽  
pp. 283
Author(s):  
M. P. Cervantes ◽  
M. Anzar ◽  
R. J. Mapletoft ◽  
J. M. Palomino ◽  
G. P. Adams
Keyword(s):  
In Vitro Maturation ◽  
Transvaginal Ultrasound ◽  
Calf Serum ◽  
Cumulus Cell ◽  
Germinal Vesicle ◽  
Bison Bison ◽  
Nuclear Maturation ◽  
Wood Bison

Methods of producing wood bison embryos in vivo and in vitro are being developed in an effort to preserve the genetic diversity of this threatened species. Previous data from our laboratory suggest that oocytes collected 24 h after LH treatment had not yet achieved nuclear maturation. The objectives of this study were (1) to determine the optimal interval of time after hCG treatment required for in vivo maturation of cumulus–oocyte complexes (COC) in wood bison, and (2) to compare the maturational characteristics of COC after in vitro v. in vivo maturation. Follicular wave emergence was synchronized among bison cows (n = 25) by follicular ablation (Day –1) from May to June. Ovarian superstimulation was induced with FSH IM diluted in 5 mg mL–1 of hyaluronan (MAP-5, Bioniche, Belleville, Ontario, Canada) given on Day 0 (300 mg) and Day 2 (100 mg). Superstimulated cows were assigned randomly to 5 groups (n = 5/group): COC collected on Day 4 with no maturation (control), or matured in vitro for 24 or 30 h, or collected 24 or 30 h after treatment with 2000 IU of hCG IM on Day 4. The COC were collected by transvaginal ultrasound-guided follicle aspiration. In vitro maturation was done in TCM-199 with 5% calf serum, 5 µg mL–1 of LH, 0.5 µg mL–1 of FSH, and 0.05 µg mL–1 of gentamicin, at 38.5°C and in 5% CO2. To assess nuclear maturation, oocytes were stained with anti-lamin AC/DAPI (4′,6-diamidino-2-phenylindole). Nuclear stages were classified as germinal vesicle (GV), GV breakdown (GVBD), metaphase I (MI), or metaphase II (MII). Comparisons among groups were made by ANOVA and Fisher’s exact test (Table 1). A mean (± SEM) of 7.6 ± 0.6 COC was collected per bison; no differences were observed among groups (P = 0.37). Cumulus cell expansion was more extensive after in vivo than in vitro maturation, and the percentage of fully expanded COC was highest in the in vivo 30-h group (97%; P < 0.05). No COC were expanded in the control (0 h) group, and none reached MI. Maximal nuclear maturation was achieved in vitro by 24 h; that is, there was no difference in the proportion of MII-stage COC at 24 versus 30 h. However, between 24 and 30 h of in vivo maturation, the percentage of nuclear stages GV + GVBD decreased from 54 to 24% (P < 0.05), whereas nuclear stages MI + MII increased from 39 to 74% (P < 0.05). In conclusion, nuclear maturation occurred earlier in vitro versus in vivo, but the consequences of this difference are unknown. Although more than one-third of oocytes matured in vivo for 30 h were mature enough to permit immediate IVF, whether additional in vivo maturation time would be beneficial to fertilization rates remains to be tested. Table 1.Nuclear status of wood bison oocytes after in vitro or in vivo maturation Thanks to Bioniche Canada.

scholarly journals INFLUENCE OF HORMONES AND FOLLICULAR FLUID ON MATURATION OF PIG OOCYTES

2001 ◽  
Vol 31 (1) ◽  
pp. 99-104
Author(s):  
Margot Alves Nunes Dode ◽  
Charles Graves
Keyword(s):  
Follicular Fluid ◽  
In Vitro Maturation ◽  
Calf Serum ◽  
Lower Percentage ◽  
Nuclear Maturation ◽  
Maturation In Vitro ◽  
Maturation Medium ◽  
Cytoplasmic Maturation ◽  
Pig Oocyte

To evaluate the effect of follicular fluid on in vitro maturation, pig oocytes were cultured in the presence of hormones where 10% of fetal calf serum (FCS), 10% of follicular fluid from large follicles (l-pFF), 10% of follicular fluid from medium follicles (m-pFF) or no supplement were added. When oocytes where matured in medium containing the hormones the addition of different supplements did not affect (P<0.05) nuclear maturation. However, changing the supplement altered the cytoplasmic maturation, with higher rate (P<0.05) observed in the l-pFF group. To determine the effect of the presence of hormone and/or supplement during maturation, the oocytes were cultured either in presence of TCM-199 alone, with hormones, with 10% l-pFF or with hormones and 10% l-pFF. The highest proportion of oocytes undergoing nuclear and cytoplasmic maturation was obtained when both hormones and follicular fluid were present. Cumulus expansion had a significant (P<0.05) effect on cytoplasmic maturation with the non-expanded groups showing a lower percentage of maturation in all groups. When the adequacy of gonadotropins levels were evaluated by adding higher or lower concentrations into the maturation medium neither beneficial nor detrimental effects were observed in either nuclear or cytoplasmic maturation. These results suggest that changes in the composition of the medium can alter the percentage of oocytes completing maturation. Follicular fluid combined with hormones was found to give better conditions for pig oocyte maturation in vitro.

scholarly journals Effect of Alpha-Tocopherol Supplementation in TCM199 Medium on In Vitro Maturation and Cleavage of Buffalo Oocytes

2020 ◽  
Vol 15 (04) ◽  
pp. 66-70
Author(s):  
Vijayalakshmi . ◽  
Shrikant Kulkarni ◽  
KB Sathisha ◽  
HM Yathish ◽  
MH Girish ◽  
...  
Keyword(s):  
In Vitro Maturation ◽  
Culture Medium ◽  
Alpha Tocopherol ◽  
Cleavage Rate ◽  
Nuclear Maturation ◽  
Maturation Rate ◽  
The Mean ◽  
Cytoplasmic Maturation ◽  
T1 And T2

An experiment was conducted to assess the effect of supplementing α-tocopherol at two different concentrations to the in vitro maturation (IVM) medium (TCM 199 culture medium) on in vitro maturation and cleavage of buffalo oocytes. Ovaries were collected from local slaughter house. Oocytes were collected by aspiration method and were matured in IVM medium (T0, control), IVM medium supplemented with α-tocopherol @ 10 μg/ml (T1) and IVM medium supplemented with α-tocopherol @ 20 μg/ml (T2). The mean cytoplasmic maturation rate was 82.39 ± 0.81, 88.81 ± 1.08, 81.67 ± 1.82 % and nuclear maturation rate was 62.50 ± 8.33, 87.50 ± 5.59, 66.66 ± 7.68 % in T0, T1 and T2 groups, respectively. The fertilized oocytes reaching 2-cell and 4-cell stages of cleavage for T0, T1 and T2 groups were 14.92 ± 1.52, 32.39 ± 1.01 and 16.39 ± 1.25 %, respectively. Significantly (p less than 0.05) higher level of cytoplasmic maturation rate, nuclear maturation rate and cleavage rate was observed in IVM medium supplemented with α-tocopherol @10 μg/ml than other two groups.

scholarly journals 301 EFFECT OF ENERGY SUBSTRATES ON METABOLISM, NUCLEAR MATURATION, AND DEVELOPMENT OF GILT AND SOW OOCYTES DURING IN VITRO MATURATION

2005 ◽  
Vol 17 (2) ◽  
pp. 301 ◽  
Author(s):  
L. Tubman ◽  
A. Peter ◽  
R. Krisher
Keyword(s):  
Amino Acids ◽  
Fatty Acids ◽  
Embryonic Development ◽  
In Vitro Maturation ◽  
Positive Control ◽  
Developmental Competence ◽  
Energy Substrates ◽  
Nuclear Maturation ◽  
Cytoplasmic Maturation

Metabolic mechanisms control both nuclear and cytoplasmic maturation in oocytes. Elevated glucose metabolism is typically associated with improved developmental competence. The objective of this study was to compare nuclear maturation, oocyte metabolism, and subsequent embryonic development following the use of different energy substrates during in vitro maturation (IVM) and to determine the specific role of each substrate. Cumulus-oocyte complexes (20–50/treatment (Trt)/replicate) were placed into maturation medium for 42 h in 7% CO2 in air at 38°C. Maturation treatments included a negative control (NC; 0.01 mM pyruvate and 6 mM lactate), addition of 1:100 dilution of fatty acids (FA; Gibco, Grand Island, NY, USA), 1 × NEAA/0.5 × EAA/1 mM glutamine (AA), or 2 mM glucose (GLU) individually; and a positive control (PC; addition of all three substrates). For each of six replicates, metabolism of 10 denuded oocytes/treatment was measured in hanging drops containing labeled glucose (0.0125 mM 5-3H glucose, glycolysis; 0.482 mM 1-14C glucose, pentose phosphate pathway, PPP). Oocytes were then fixed and stained for determination of meiotic stage. Remaining oocytes were fertilized and cultured in vitro. Cleavage and blastocyst development were recorded at 30–40 and 144 h post-insemination, respectively. The Purdue Porcine Media system was used throughout (PPM; Herrick et al. 2003 Reprod. Fertil. Dev. 15, 249–254). All data were subjected to analysis of variance. Oocyte metabolism and embryonic development are presented In Table 1. Except for FA, energy substrate influenced the percentage of oocytes reaching metaphase II (NC, 1.37 ± 0.01; FA, 1.35 ± 0.01; AA, 33.33 ± 0.06; GLU, 25.81 ± 0.06; PC, 54.29 ± 0.06) but age of oocyte donor did not. Blastocyst metabolism and cell number were not affected by treatment. In general, sows were more responsive to treatment effects. These data demonstrate that exogenous fatty acids do not play a role in porcine oocyte maturation. Amino acids appear to promote meiosis and glycolysis, but do not support oocyte developmental potential. Elevated metabolism in this treatment may be due to a recovery effect when glucose-starved oocytes were placed into glucose containing metabolism medium. Glucose appears to be important for meiosis and cytoplasmic maturation leading to developmental competence with minimal effect on oocyte metabolism. The success of the positive control suggests that a combination of glucose and amino acids is beneficial to maturation and embryonic development of porcine oocytes. Table 1. Metabolism and development of oocytes after IVM

scholarly journals Cyclic nucleotide in oocyte In vitro maturation in Assisted Reproductive Technology

2020 ◽  
Vol 6 (3) ◽  
pp. 110-120
Author(s):  
Christie L Sun ◽  
Sally L Catt ◽  
Kiri Beilby ◽  
Mulyoto Pangestu
Keyword(s):  
Assisted Reproductive Technology ◽  
Natriuretic Peptide ◽  
In Vitro Maturation ◽  
Reproductive Technology ◽  
Developmental Competence ◽  
Nuclear Maturation ◽  
Oocyte Developmental Competence ◽  
Cytoplasmic Maturation ◽  
Camp Levels

In vitro maturation (IVM) is a promising assisted reproductive technology (ART) for human infertility treatment. However, when cumulus oocyte complexes (COCs) are removed from their follicular environment when manipulated in vitro, it can lead to a decrease of intra-oocyte cyclic adenosine 3’, 5’-monophosphare (cAMP) causing spontaneous nuclear maturation and an asynchrony with the oocytes’ cytoplasmic maturation, resulting in poor embryo developmental outcomes. Nuclear and cytoplasmic synchrony is important during oocyte maturation within antral follicles.It is maintained partially by the actions of c-type natriuretic peptide (CNP) binding with natriuretic peptide receptor 2 (NPR2), supporting high cAMP levels thus holding the oocyte in meiotic arrest. Addition of CNP to pre-IVM media has the capacity of maintaining cAMP levels and thus improve synchrony. Moreover, in women with advanced maternal age, successful IVM of aging oocytes faces significant challenges due to the morphological and cellular changes.  Inhibiting initiation of nuclear maturation by cAMP modulator, CNP during pre-IVM period and thus improve oocyte developmental competence regardless of oocyte age.

Sign in / Sign up

Export Citation Format

Share Document