Recovery of mitochondrial function and endogenous antioxidant systems in vitrified bovine oocytes during extended in vitro culture

2011 ◽  
Vol 78 (12) ◽  
pp. 942-950 ◽  
Author(s):  
Xue-Ming Zhao ◽  
Wei-Hua Du ◽  
Dong Wang ◽  
Hai-Sheng Hao ◽  
Yan Liu ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Mitochondrial Function ◽  
Antioxidant Systems ◽  
Endogenous Antioxidant ◽  
Bovine Oocytes

In vitro culture of secondary follicles and prematuration of cumulus–oocyte complexes from antral follicles increase the levels of maturation‐related transcripts in bovine oocytes

2019 ◽  
Vol 86 (12) ◽  
pp. 1874-1886
Author(s):  
Francisco Taiã G. Bezerra ◽  
Francisco Edilcarlos O. Lima ◽  
Laís Rayani F. M. Paulino ◽  
Bianca R. Silva ◽  
Anderson W. B. Silva ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Antral Follicles ◽  
Bovine Oocytes

242 USE OF BRAIN-DERIVED NEUROTROPHIC FACTOR IN IN VITRO PREMATURATION OF BOVINE OOCYTES SUBJECTED TO PARTHENOGENETIC ACTIVATION

2008 ◽  
Vol 20 (1) ◽  
pp. 200
Author(s):  
T. H. C. De Bem ◽  
R. Rochetti ◽  
P. R. L. Pires ◽  
F. F. Bressan ◽  
P. R. Adona ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Embryo Development ◽  
Polar Body ◽  
Hatching Rate ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Parthenogenetic Activation ◽  
Bovine Oocytes ◽  
Hatching Rates

Prematuration provides an additional time for oocyte capacitation and maturation in an attempt to improve in vitro embryo production (IVP) rates and allows media supplementation during this period for IVP. The aim of this study was to use brain-derived neurotropic factor (BDNF) in prematuration to improve maturation of bovine oocytes subjected to parthenogenetic activation and cultured with different media. Oocytes were subjected to prematuration in TCM-199 medium supplemented with 10 µm butyrolactone I, 2.0 mm pyruvate, and 10 µg mL–1 gentamicin for 24 h in the absence of BDNF (control) or in the presence of 10 ng mL–1 BDNF (BD). Oocytes were then in vitro-matured (IVM) in TCM-199 medium supplemented with 10% FCS, 0.5 µg mL–1 FSH, 5.0 µg mL–1 LH, 2.0 mm pyruvate, and 10 µg mL–1 gentamicin at 38.5�C under 5% CO2 in air. After 19 h oocytes were denuded using hyaluronidase and vortexing for 3 min for the 1st polar body (1PB) selection. Those which extruded the 1PB were maintained in IVM until 26 h, when parthenogenetic activation was performed (5 min in 5 µm ionomycin, followed by 3 h in 2 mm 6-DMAP). Activated oocytes were then transferred to in vitro culture (IVC) for embryo development evaluation. Embryos from both groups were cultured in SOF medium with 2.5% FCS, 0.05 g mL–1 BSA, 0.2 mm pyruvate, and 10 mg mL–1 gentamicin. Cleavage rates on the second day of in vitro culture (D2), embryo production at Days 7 and 8 (D7 and D8), and hatching rate at Day 8 were evaluated. Data regarding 1PB extrusion, cleavage, blastocyst development on D7 and D8, and blastocyst D8 hatching rates of three replicates were analyzed by chi-square test at 5% significance using the BIOESTATS 4.0 software. Control and BD, respectively, did not show differences (P > 0.05) regarding 1PB extrusion (n = 164, 63.81%, and n = 175, 66.79%) or cleavage (n = 117, 71.34%, and n = 138, 78.86%). However, for control and BD, respectively, blastocyst development on D7 (n = 63, 38.41%, and n = 89, 50.86%), D8 (n = 63, 38.41%, and n = 91, 52.00%), and hatching on D8 (n = 22, 34.92%, and n = 39, 43.82%) were all significantly higher for BD when compared with control (P < 0.05). In conclusion, BDNF during prematuration improved in vitro embryo development by increasing blastocyst and hatching rates of parthenogenetic embryos.

174 Effects of Melatonin on In Vitro Maturation of Bovine Oocytes and Gene Expression in Cumulus Cells: Preliminary Results

2018 ◽  
Vol 30 (1) ◽  
pp. 226
Author(s):  
F. C. Castro ◽  
L. Schefer ◽  
K. L. Schwarz ◽  
H. Fernandes ◽  
R. C. Botigelli ◽  
...  
Keyword(s):  
Gene Expression ◽  
In Vitro Culture ◽  
Reverse Transcription ◽  
In Vitro Maturation ◽  
Cumulus Cells ◽  
Culture Conditions ◽  
Nuclear Maturation ◽  
Maturation Rate ◽  
Bovine Oocytes

Melatonin mediates several processes in animal reproduction and has drawn attention for its potent antioxidant, anti-apoptotic, anti-inflammatory action and, more recently, for its benefits on oocyte maturation and embryo development in vitro. The aim of this study was to assess the effect of melatonin during the in vitro maturation (IVM) on nuclear maturation of bovine oocytes and gene expression in their corresponding cumulus cells (CC). Bovine cumulus–oocyte complexes (COC) were obtained by aspiration of follicles (2-6 mm) from slaughterhouse ovaries, selected (grades I and II) and transferred to 4 well plates (25-30 COC/well) containing IVM medium [TCM-199 supplemented with sodium bicarbonate (26 mM), sodium pyruvate (0.25 mM), FSH (0.5 µg mL−1), LH (5.0 µg mL−1), 0.3% BSA, and gentamicin (50 µg mL−1)] with 0, 10−5, 10−7, 10−9 or 10−11 M melatonin and cultured for 24 h at 38.5°C and 5% CO2. At the end of IVM, oocytes were stained with Hoechst 33342 (10 μg mL−1) and evaluated for nuclear maturation rate. The CC were evaluated for the expression of antioxidant (SOD1, SOD2, GPX4), pro-apoptotic (P53, BAX) and expansion-related genes (PTX3, HAS1, HAS2). For transcript detection in CC, RNA isolation was performed with TRIzol®Reagent (Invitrogen, Carlsbad, CA, USA) and reverse transcription with High Capacity cDNA Reverse Transcription kit (Applied Biosystems, Foster City, CA, USA). Relative quantification of transcripts was performed by RT-qPCR using 3 endogenous controls (β-actin, GAPDH, PPIA). Nuclear maturation rate and gene expression were tested by ANOVA and means were compared by Tukey’s test (6 replicates). In CC, the different concentrations of melatonin did not significantly alter expression of the investigated genes (P > 0.05), although all concentrations provided a numerical increase in the expression of the antioxidant SOD1 and of the expansion-related genes PTX3 and HAS2. Regarding the pro-apoptotic genes, concentrations of 10−11 and 10−9 M were able to reduce only numerically the expression of BAX and P53, respectively. In oocytes, the rate of nuclear maturation was not different among the tested treatments (P > 0.05), but it was numerically higher in the 10−7 M melatonin treated group compared with the control (69.71 ± 13.76% v. 88.1 ± 12.54%). In conclusion, under the studied conditions, melatonin was unable to improve maturation rate or to affect the expression of antioxidant, pro-apoptotic, and expansion-related genes in CC. Melatonin during IVM has shown variable results in different studies and appears to show different effects depending on culture conditions and parameters studied. In order to take advantage of the possible positive antioxidant effects of melatonin, other culture conditions and parameters should be investigated. In a next step, melatonin will be included during in vitro culture of embryos to evaluate its possible cytoprotective role, because such embryos are more exposed to oxidative stress during in vitro culture, and to investigate its benefits on developmental competence in vitro. This reaesrch was funded by FAPESP (2015/20379-0; 2014/17181-0).

scholarly journals Effect of Cooking Methods on the Antioxidant Capacity of Plant Foods Submitted to In Vitro Digestion–Fermentation

Antioxidants ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1312
Author(s):  
Beatriz Navajas-Porras ◽  
Sergio Pérez-Burillo ◽  
Álvaro Jesús Valverde-Moya ◽  
Daniel Hinojosa-Nogueira ◽  
Silvia Pastoriza ◽  
...  
Keyword(s):  
Antioxidant Capacity ◽  
Thermal Processing ◽  
In Vitro Digestion ◽  
Antioxidant Systems ◽  
Cooking Methods ◽  
Antioxidant Compounds ◽  
Plant Foods ◽  
Total Antioxidant ◽  
Endogenous Antioxidant

The antioxidant capacity of foods is essential to complement the body’s own endogenous antioxidant systems. The main antioxidant foods in the regular diet are those of plant origin. Although every kind of food has a different antioxidant capacity, thermal processing or cooking methods also play a role. In this work, the antioxidant capacity of 42 foods of vegetable origin was evaluated after in vitro digestion and fermentation. All foods were studied both raw and after different thermal processing methods, such as boiling, grilling roasting, frying, toasting and brewing. The cooking methods had an impact on the antioxidant capacity of the digested and fermented fractions, allowing the release and transformation of antioxidant compounds. In general, the fermented fraction accounted for up to 80–98% of the total antioxidant capacity. The most antioxidant foods were cocoa and legumes, which contributed to 20% of the daily antioxidant capacity intake. Finally, it was found that the antioxidant capacity of the studied foods was much higher than those reported by other authors since digestion–fermentation pretreatment allows for a higher extraction of antioxidant compounds and their transformation by the gut microbiota.

256 SYNERGISTIC EFFECT ON EMBRYO DEVELOPMENT AND EMBRYO QUALITY WHEN BOVINE DENUDED OOCYTES ARE CO-CULTURED WITH CUMULUS - OOCYTE COMPLEXES DURING IN VITRO MATURATION

2011 ◽  
Vol 23 (1) ◽  
pp. 226
Author(s):  
S. R. Dey ◽  
G. K. Deb ◽  
J. I. Bang ◽  
S. J. Cho ◽  
B. H. Choi ◽  
...  
Keyword(s):  
Synergistic Effect ◽  
In Vitro Culture ◽  
In Vitro Maturation ◽  
Cumulus Cell ◽  
Tunel Staining ◽  
Oocyte Growth ◽  
Respective Control ◽  
Bovine Oocytes

The oocyte and its surrounding somatic cells are metabolically coupled to each other through gap junctions. This phenomenon allows intercellular communication and transfer of different low-molecular-weight substrates between the cells necessary for oocyte growth. The oocyte itself regulates the cumulus cell microenvironment through oocyte-secreted factors. The development competence of the bovine oocytes is increased when denuded oocytes (DO) are co-cultured with cumulus–oocyte complexes (COC) during in vitro maturation (IVM). However, the fate of the DO, which are usually discarded after IVM, has not been determined. The present study aimed to investigate whether there is a synergistic effect of co-culturing COC and DO during IVM. We performed 3 IVM schemes: 1) COC and DO co-culture, with 12 COC and 60 DO; 2) COC control, with 12 COC; and 3) DO control, with 60 DO in 120-μL drop of TCM-199 for 22 to 24 h. Following IVM, IVF and in vitro culture were separately performed for the COC (COC co-culture) and DO (DO co-culture) from the IVM co-culture group. In vitro fertilization and in vitro culture (modified CR1aa) were done in 60-μL drops. Embryos were cultured at 38.5°C and 5% CO2 in air. Cleavage and blastocyst rates were checked at Day 3 and 8 from IVF on total COC/DO placed in IVM drop. Day 8 blastocysts were used for TUNEL staining using In Situ Cell Death Detection Kit (Roche, Budapest, Hungary). Data were analysed by one-way ANOVA, and significant differences among groups were tested by DMRT. Compared with the respective control treatments, co-culture has no effect on cleavage rates of COC and DO (see Table 1). However, blastocyst rates and total cell numbers of blastocysts were increased in COC co-culture and DO co-culture group compared with their respective control groups (see Table 1). Co-culture had no effect on apoptosis of blastocysts. These data show that co-culture of COC and DO improved developmental competence and quality of embryos from the COC co-culture and DO co-culture group. Table 1.Development competence and blastocyst quality of intact and denuded bovine oocytes This work was partly supported by the BK21 program, the KRF (KRF-2008-211-F00011), the IPET (108068-03-1-SB010), and the KOSEF (10525010001-05N2501-00110).

Oxidative stress regulates collagen synthesis and matrix metalloproteinase activity in cardiac fibroblasts

2001 ◽  
Vol 280 (1) ◽  
pp. C53-C60 ◽  
Author(s):  
Deborah A. Siwik ◽  
Patrick J. Pagano ◽  
Wilson S. Colucci
Keyword(s):  
Oxidative Stress ◽  
Extracellular Matrix ◽  
Collagen Synthesis ◽  
Cardiac Fibroblasts ◽  
Antioxidant Systems ◽  
Leucine Incorporation ◽  
Gelatinase Activity ◽  
Adult Rat ◽  
Endogenous Antioxidant

Oxidative stress has been implicated in the pathophysiology of myocardial failure. We tested the hypothesis that oxidative stress can regulate extracellular matrix in cardiac fibroblasts. Neonatal and adult rat cardiac fibroblasts in vitro were exposed to H2O2 (0.05–5 μM) or the superoxide-generating system xanthine (500 μM) plus xanthine oxidase (0.001–0.1 mU/ml) (XXO) for 24 h. In-gel zymography demonstrated that H2O2 and XXO each increased gelatinase activity corresponding to matrix metalloproteinases (MMP) MMP-13, MMP-2, and MMP-9. H2O2 and XXO decreased collagen synthesis (collagenase-sensitive [3H]proline incorporation) without affecting total protein synthesis ([3H]leucine incorporation). H2O2 and XXO decreased the expression of procollagen α1(I), α2(I), and α1(III) mRNA but increased the expression of fibronectin mRNA, suggesting a selective transcriptional effect on collagen synthesis. H2O2, but not XXO, also decreased the expression of nonfibrillar procollagen α1(IV) and α2(IV) mRNA. To determine the role of endogenous antioxidant systems, cells were treated with the superoxide dismutase (SOD) inhibitor diethyldithiocarbamic acid (DDC, 100 μM) to increase intracellular superoxide or with the glucose-6-phosphate dehydrogenase inhibitor dehydroisoandrosterone 3-acetate (DHEA; 10 μM) to increase intracellular H2O2. DDC and DHEA decreased collagen synthesis and increased MMP activity, and both effects were inhibited by an SOD/catalase mimetic. Thus increased oxidative stress activates MMPs and decreases fibrillar collagen synthesis in cardiac fibroblasts. Oxidative stress may play a role in the pathogenesis of myocardial remodeling by regulating the quantity and quality of extracellular matrix.

scholarly journals Effect of Cooking Methods on the Antioxidant Capacity of Foods of Animal Origin Submitted to In Vitro Digestion-Fermentation

Antioxidants ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 445
Author(s):  
Beatriz Navajas-Porras ◽  
Sergio Pérez-Burillo ◽  
Álvaro Valverde-Moya ◽  
Daniel Hinojosa-Nogueira ◽  
Silvia Pastoriza ◽  
...  
Keyword(s):  
Antioxidant Capacity ◽  
In Vitro Digestion ◽  
Bioactive Molecules ◽  
Animal Origin ◽  
Antioxidant Systems ◽  
Cooking Methods ◽  
Antioxidant Compounds ◽  
Microbial Fermentation ◽  
Endogenous Antioxidant

The human body is exposed to oxidative damage to cells and though it has some endogenous antioxidant systems, we still need to take antioxidants from our diet. The main dietary source of antioxidants is vegetables due to their content of different bioactive molecules. However, there are usually other components of the diet, such as foods of animal origin, that are not often linked to antioxidant capacity. Still, these foods are bound to exert some antioxidant capacity thanks to molecules released during gastrointestinal digestion and gut microbial fermentation. In this work, the antioxidant capacity of 11 foods of animal origin has been studied, submitted to different culinary techniques and to an in vitro digestion and gut microbial fermentation. Results have shown how dairy products potentially provide the highest antioxidant capacity, contributing to 60% of the daily antioxidant capacity intake. On the other hand, most of the antioxidant capacity was released during gut microbial fermentation (90–98% of the total antioxidant capacity). Finally, it was found that the antioxidant capacity of the studied foods was much higher than that reported by other authors. A possible explanation is that digestion–fermentation pretreatment allows for a higher extraction of antioxidant compounds and their transformation by the gut microbiota. Therefore, although foods of animal origin cannot be compared to vegetables in the concentration of antioxidant molecules, the processes of digestion and fermentation can provide some, giving animal origin food some qualities that could have been previously unappreciated.

223 RELATIONSHIP BETWEEN THE LENGTH OF CELL CYCLES, CLEAVAGE PATTERN, AND DEVELOPMENTAL COMPETENCE DURING IN VITRO CULTURE OF IN VITRO-MATURED/IN VITRO-FERTILIZED BOVINE OOCYTES

2009 ◽  
Vol 21 (1) ◽  
pp. 209 ◽  
Author(s):  
T. Somfai ◽  
Y. Inaba ◽  
Y. Aikawa ◽  
M. Ohtake ◽  
S. Kobayashi ◽  
...  
Keyword(s):  
Cell Division ◽  
In Vitro Culture ◽  
Cell Mass ◽  
Calf Serum ◽  
Blastocyst Stage ◽  
Developmental Competence ◽  
Cleavage Pattern ◽  
Cell Cycles ◽  
Bovine Oocytes

In in vitro embryo production systems, there is a need to select embryos with good developmental competence at the early stages. This study was conducted to determine whether there was any relationship between the duration of the first 3 cell cycles, the cleavage pattern of the first cell division, and the developmental competence of embryos during in vitro culture. A total of 320 in vitro-matured and in vitro-fertilized bovine oocytes were cultured in microdrops of CR1aa medium supplemented with 5% calf serum covered by mineral oil in 5% CO2 and 20% O2 at 38.5°C. The kinetics of embryo development were measured by time-lapse cinematography. Embryos were classified according to their cleavage pattern at the first cell division. Of 285 cleaved embryos, 119 had 2 blastomeres of the same size (normal cleavage: NC), 49 had 2 blastomeres with multiple small fragments (multiple fragments: MF), 34 had 2 blastomeres and a protrusion (protrusion: PT), 45 showed direct cleavage from 1 cell to 3 or 4 blastomeres (3–4BL), and 60 oocytes cleaved to 2 blastomeres of different sizes (unequal blastomeres: UB). (Twenty-two embryos belonged to 2 classes.) After 175 h of culture, blastocysts were either subjected to differential inner cell mass/trophectoderm (ICM/TE) staining or karyotyped. The first and second cell cycles (mean ± SEM) of viable embryos (that could develop to the blastocyst stage) were significantly shorter than those of nonviable embryos (24.9 ± 0.3 h and 8.7 ± 0.1 h v. 26.6 ± 0.7 h and 10.0 ± 0.1 h, respectively); however, the length of the third cell cycle did not differ (P < 0.05, paired t-test). The duration of 1 cell stage in the NC group was significantly shorter than that of MF, PT, 3–4BL, and UB groups (24.7 ± 0.4 h, 26.6 ± 0.5 h, 26.3 ± 0.6 h, 26.0 ± 0.2 h, and 27.7 ± 0.9 h, respectively). The length of the second and third cell cycles did not differ among the groups. The percentage of NC embryos that developed to the blastocyst stage was similar to that of the 3–4BL group (66.9 and 56.7%, respectively) but was significantly higher than those of the MF, PT, and UB groups (40.5, 26.5, and 35.6%, respectively; P < 0.05, ANOVA). The mean cell numbers of NC blastocysts did not differ from those of the MF, 3–4BL, and UB groups but were higher than those of PT embryos (147.1, 155.6, 121.6, 146.4, and 115.1, respectively). There was no difference in ICM/TE rates between the groups. Unlike NC, MF, PT, and UB embryos, most (6 of 8 karyotyped) 3–4BL blastocysts had abnormal ploidy, such as haploid, triploid, mixoploid, or chaotic chromosome numbers, in blastomeres. Our results revealed that not only the length of the first cell cycles, but also the cleavage pattern during first cell division can be a marker of developmental competence and should be considered for the selection of good-quality embryos for embryo transfer. This work was supported by the Research and Development Program for New Bio-industry Initiatives.

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