33 Effect of extended culture after vitrification-warming of bovine oocytes on mitochondrial function

2022 ◽  
Vol 34 (2) ◽  
pp. 251
Author(s):  
E. J. Gutierrez-Castillo ◽  
S. A. Talbot ◽  
F. A. Diaz ◽  
K. R. Bondioli
Keyword(s):  
Mitochondrial Function ◽  
Extended Culture ◽  
Bovine Oocytes

193 INSULIN-LIKE GROWTH FACTOR-1 EXERTS A THERMOPROTECTIVE ROLE ON MITOCHONDRIAL FUNCTION OF BOVINE OOCYTES EXPOSED TO HEAT SHOCK

2012 ◽  
Vol 24 (1) ◽  
pp. 209 ◽  
Author(s):  
J. Ispada ◽  
R. S. Lima ◽  
P. H. B Risolia ◽  
M. E. O. A. Assumpção ◽  
J. A. Visintin ◽  
...  
Keyword(s):  
Elevated Temperature ◽  
Growth Factor ◽  
Heat Shock ◽  
Mitochondrial Function ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Insulin Like Growth Factor ◽  
Bovine Oocyte ◽  
Bovine Oocytes ◽  
Apoptotic Cascade

The series of events associated with oocyte maturation are susceptible to disruption by elevated temperature. These events are regulated by a variety of growth factors, such as insulin-like growth factor-1 (IGF-1). Exposure of bovine oocytes to heat shock compromises oocyte competence and triggers apoptosis. It has been shown that cellular stresses often alter mitochondrial function and activate the mitochondrial apoptotic cascade. Therefore, the objective of this study was to determine the effect of heat shock on bovine oocyte mitochondrial activity and the role of IGF-1 in this context. Slaughterhouse derived cumulus–oocyte complexes (COC) were subjected to control (38.5°C for 22 h) and heat shock (41°C for 14 h, followed by 38.5°C for 8 h) treatments in the presence of 0 or 100 ng mL–1 of IGF-1 during in vitro maturation (IVM). After 22 h, IVM COC were mechanically denuded and subjected to MitoTracker Red CMX-Ros assay (Invitrogen M-7512) to localize and quantify active mitochondria. Denuded oocytes were incubated in TCM-199-HEPES containing 10 μg mL–1 of polyvinyl alcohol and 50 nM MitoTracker at 37°C for 15 min. Oocytes were evaluated under fluorescence microscope and digital images were obtained and stored as TIFF files. Mitochondrial activity from each oocyte was quantified using the software Image J 1.43. This experiment was replicated 6 times using 97 to 204 COC/treatment. Data were analyzed by least-squares analysis of variance using the general linear model procedure of SAS. In the absence of IGF-1, heat shock reduced (P < 0.001) mitochondrial activity from 64.31 ± 1.91 to 56.74 ± 1.26 arbitrary units for control and heat shock groups, respectively. Addition of IGF-1 to maturation medium did not affect mitochondrial activity in the control group (66.25 ± 1.56). However, IGF-1 improved (temperature × IGF-1; P < 0.001) mitochondrial activity of bovine oocytes subjected to heat shock (70.32 ± 1.32). In conclusion, heat shock reduced bovine oocyte mitochondrial activity, suggesting activation of mitochondrial apoptotic cascade. Moreover, IGF-1 exerted a thermoprotective role, reducing the mitochondrial damage caused by elevated temperature.

36 Extended culture after vitrification-warming helps in spindle recovery of bovine oocytes

2020 ◽  
Vol 32 (2) ◽  
pp. 144
Author(s):  
E. Gutierrez ◽  
Z. Jiang ◽  
K. Bondioli
Keyword(s):  
Ethylene Glycol ◽  
Dimethyl Sulfoxide ◽  
Incubation Period ◽  
The Other ◽  
Meiotic Spindle ◽  
Chromosome Arrangement ◽  
Extended Culture ◽  
Base Medium ◽  
Maturation Medium ◽  
Bovine Oocytes

The meiotic spindle is one of the most vulnerable cytoplasmic organelles when performing oocyte vitrification. It has been proposed that submitting oocytes to a post-warming incubation period in maturation medium helps in the reorganization of microtubules and chromosomes. Our previous experiments found no differences in spindle morphology after submitting vitrified oocytes to a 2-h incubation period. The aim of this experiment was to determine the effect of extended culture on the reorganization of the meiotic spindle of vitrified-warmed bovine oocytes. Oocytes were purchased from a commercial vendor (n=86) and matured during shipment. In this experiment, three treatments were evaluated: fresh oocytes (F) (n=30), vitrified-warmed (VW; n=26), and extended culture (EC; n=30). Cumulus-oocyte complexes were removed at 18h of maturation. Fresh oocytes were denuded by vortexing in hyaluronidase (1.5mgmL−1) and immediately fixed using 4% paraformaldehyde. Oocytes undergoing vitrification were partially denuded by pipetting in hyaluronidase (1.5mgmL−1). The vitrification protocol consisted of incubation in equilibration solution (7.5% dimethyl sulfoxide + 7.5% ethylene glycol) for 9min and then in vitrification solution (15% dimethyl sulfoxide + 15% ethylene glycol + 0.5M sucrose). While in vitrification solution, oocytes were mounted onto a Cryolock and plunged into liquid nitrogen in less than 1min. Warming was performed by placing a Cryolock into 0.5M sucrose for 3min and then into 0.25M sucrose for 3min. Finally, oocytes were washed in base medium. The base medium used for cryoprotectant and warming solutions was Dulbecco's phosphate-buffered saline supplemented with 20% fetal bovine serum. Both, vitrification and warming, were performed at 38.5°C. After warming, half of the oocytes were completely denuded and fixed and the other half underwent a 6-h incubation period in maturation medium (IVF-Bioscience). To examine microtubule distribution and chromosome arrangement, fixed oocytes were submitted to an immunostaining protocol using α tubulin antibody (1:100) and anti IgG-Alexa Fluor 488 (1:1000; Thermo Fisher Scientific) and counterstained with Hoechst. The effect of extended culture on the incidence of abnormal microtubule distribution and chromosome arrangement was analysed using logistic regression with a binomial response variable (normal/abnormal). There was no difference in maturation rates among groups (F=73.3%, VW=77%, EC=86.6%; P=0.43). For microtubule distribution, oocytes fixed immediately after warming had a higher incidence of abnormal spindles (57.7%) when compared with oocytes submitted to extended culture (26.6%; P=0.02). The most common abnormality seen in oocytes fixed after warming was small and faintly stained spindles. Microtubule distribution in fresh oocytes did not differ from oocytes in the other groups. There were no differences in chromosome arrangement among groups (P=0.11). Future research will focus on evaluating the benefits that this technique offers to improve development following IVF using vitrified-warmed oocytes.

195 HEAT STRESS ALTERS THE TRANSCRIPTOME OF MATURING BOVINE OOCYTES

2016 ◽  
Vol 28 (2) ◽  
pp. 228
Author(s):  
L. A. Rispoli ◽  
R. R. Payton ◽  
C. Gondro ◽  
A. M. Saxton ◽  
J. L. Edwards
Keyword(s):  
Protein Synthesis ◽  
Heat Stress ◽  
Elevated Temperature ◽  
Mitochondrial Function ◽  
De Novo ◽  
Bovine Genome ◽  
Rna Extraction ◽  
Developmental Competence ◽  
The Impact ◽  
Bovine Oocytes

Direct exposure of maturing oocytes to a physiologically relevant elevated temperature reduces embryo development after fertilisation and has been coincident with reduced de novo protein synthesis. Mechanisms responsible for heat-induced reductions in protein synthesis are unknown but may be related to alterations in the transcriptome of the maturing oocyte. To determine the extent to which this may occur, the impact of heat stress on the maternal pool of RNA in bovine oocytes was assessed using microarrays. After maturation for 24 h at 38.5°C (control) or 41°C (first 12 h only, 38.5°C thereafter; heat stress) oocytes were denuded from associated cumulus cells and lysed for RNA extraction or underwent IVF to assess developmental competence. Total RNA from oocytes was amplified by 3′-poly(A) priming or a combination of 3′-poly(A) and internal priming because oocyte transcripts may or may not have a polyadenylated tail. Amplified RNA was hybridised to GeneChip Bovine Genome Arrays (Affymetrix, Santa Clara, CA, USA; 8 oocyte pools per treatment were collected on 7 different occasions and amplified by 2 methods; n = 32 chips). Differential transcript abundance was determined using R and Bioconductor with only probes having a P < 0.01, a fold change of at least 1.3, and called present for at least half the arrays. Functional annotation of selected transcripts was performed using Gene Ontology and KEGG annotations (Bos taurus build 4.0) and DAVID (v 6.7) with significance level set at P < 0.10. Coincident with reduced blastocyst development (28.3 v. 15.2% for control v. heat stress, respectively; SEM = 3.6; P < 0.0003), heat stress altered the abundance of 159 transcripts (22 increased, 137 decreased); 130 of these were annotated. Use of DAVID demonstrated enrichment of genes important for mitochondrial function and RNA processing. Towards validating certain findings, the relative abundance of 3 mitochondrial transcripts (NDUFC2, COQ3, ATP5O) were assessed by quantitative PCR on non-amplified RNA from the oocyte samples used for the microarray study. Gene-specific primers were designed for 5′ and 3′ ends of transcripts when possible. Exposure to elevated temperature during the first 12 h of oocyte maturation reduced transcript levels of NDUFC2 at the 5′ and 3′ ends (P < 0.0001 and P = 0.003), COQ3 at the 3′ end (P = 0.02) and ATP5O at the 5′ end (P = 0.02). In conclusion, exposure of maturing cumulus-oocyte complexes to a physiologically-relevant elevated temperature altered the transcriptome in oocytes, especially certain transcripts important for mitochondrial function. 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.

Effect of 5-aminoimidazole-4-carboxamide ribonucleoside on the mitochondrial function and developmental ability of bovine oocytes

2015 ◽  
Vol 84 (4) ◽  
pp. 490-497 ◽  
Author(s):  
Shun Takeo ◽  
Takahito Abe ◽  
Koumei Shirasuna ◽  
Takehito Kuwayama ◽  
Hisataka Iwata
Keyword(s):  
Mitochondrial Function ◽  
Bovine Oocytes

scholarly journals Resveratrol Improves the Mitochondrial Function and Fertilization Outcome of Bovine Oocytes

2014 ◽  
Vol 60 (2) ◽  
pp. 92-99 ◽  
Author(s):  
Shun TAKEO ◽  
Daichi SATO ◽  
Koji KIMURA ◽  
Yasunori MONJI ◽  
Takehito KUWAYAMA ◽  
...  
Keyword(s):  
Mitochondrial Function ◽  
Bovine Oocytes

scholarly journals Mitochondrial function in immature bovine oocytes is improved by an increase of cellular cyclic AMP

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Shu Hashimoto ◽  
Masaya Yamanaka ◽  
Takayuki Yamochi ◽  
Hisataka Iwata ◽  
Ryouka Kawahara-Miki ◽  
...  
Keyword(s):  
Cyclic Amp ◽  
Mitochondrial Function ◽  
Bovine Oocytes

Rigorous swim training impairs mitochondrial function in post-ischaemic rat heart

1997 ◽  
Vol 160 (1) ◽  
pp. 139-148
Author(s):  
S.B. LEICHTWEIS ◽  
C. LEEUWENBURGH ◽  
D. J. PARMELEE ◽  
R. FIEBIG ◽  
L. L. JI
Keyword(s):  
Mitochondrial Function ◽  
Rat Heart

Residual hepatic mitochondrial function assessment through 13C-methionine, 13C-octanoate and 13C-ketoisocaproate breath tests

2001 ◽  
Vol 120 (5) ◽  
pp. A566-A566
Author(s):  
A ARMUZZI ◽  
M ZOCCO ◽  
M CANDELLI ◽  
C DICAMPLI ◽  
E NISTA ◽  
...  
Keyword(s):  
Mitochondrial Function ◽  
Breath Tests ◽  
Function Assessment
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