scholarly journals Effect of Embryo Density on In Vitro Development and Gene Expression in Bovine In Vitro-fertilized Embryos Cultured in a Microwell System

2013 ◽  
Vol 59 (2) ◽  
pp. 115-122 ◽  
Author(s):  
Satoshi SUGIMURA ◽  
Tomonori AKAI ◽  
Yutaka HASHIYADA ◽  
Yoshio AIKAWA ◽  
Masaki OHTAKE ◽  
...  
Keyword(s):  
Gene Expression ◽  
In Vitro Development ◽  
Vitro Development

Effects of the HDAC inhibitor scriptaid on the in vitro development of bovine embryos and on imprinting gene expression levels

2018 ◽  
Vol 110 ◽  
pp. 79-85 ◽  
Author(s):  
R. Laguna-Barraza ◽  
M.J. Sánchez-Calabuig ◽  
A. Gutiérrez-Adán ◽  
D. Rizos ◽  
S. Pérez-Cerezales
Keyword(s):  
Gene Expression ◽  
Hdac Inhibitor ◽  
Bovine Embryos ◽  
In Vitro Development ◽  
Expression Levels ◽  
Imprinting Gene ◽  
Vitro Development ◽  
Gene Expression Levels

Vitrification of bovine blastocysts pretreated with sublethal hydrostatic pressure stress: evaluation of post-thaw in vitro development and gene expression

2011 ◽  
Vol 23 (4) ◽  
pp. 585 ◽  
Author(s):  
E. Siqueira Filho ◽  
E. S. Caixeta ◽  
C. Pribenszky ◽  
M. Molnar ◽  
A. Horvath ◽  
...  
Keyword(s):  
Gene Expression ◽  
Hydrostatic Pressure ◽  
Transcript Abundance ◽  
Developmental Competence ◽  
Hatching Rate ◽  
In Vitro Development ◽  
Stress Treatment ◽  
Pressure Stress ◽  
Vitro Development

Sublethal stress treatment has been reported to enhance gametes’ performance in subsequent procedures, such as cryopreservation. The aim of the present study was to evaluate the effect of different equilibration times between the termination of a sublethal hydrostatic pressure (HP) stress treatment and the initiation of vitrification on the post-thaw survival, continued in vitro development, hatching rate and gene expression of selected candidate genes of in vitro-produced (IVP) expanded bovine blastocysts. Day 7 IVP blastocysts were subjected to 600 bar pressure for 60 min at 32°C. Immediately after pressure treatment (HP0h) or after 1 or 2 h incubation (HP1h and HP2h groups, respectively), embryos were either vitrified and warmed using the open pulled straw method, followed by 72 h in vitro culture or were stored at –80°C until gene expression analysis. Re-expansion and hatching rates after vitrification–warming were significantly (P < 0.05) higher in the HP0h (88 and 76%, respectively) and HP1h (90 and 75%, respectively) groups than in the untreated (82 and 63%, respectively) and HP2h groups (79 and 70%, respectively). Moreover, the HP1h group showed further improvement in the speed of re-expansion and resumption of normal in vitro development. Cumulative analysis of all genes (SC4MOL, HSP1A1A, SOD2 and GPX4) revealed a similar pattern of expression, with a tendency for peak transcript abundance 1 h after HP treatment. Application of HP stress treatment was found to be efficient in increasing the in vitro developmental competence of vitrified bovine embryos.

Effects of hyaluronan, BSA, and serum on bovine embryo in vitro development, ultrastructure, and gene expression patterns

2006 ◽  
Vol 73 (12) ◽  
pp. 1503-1511 ◽  
Author(s):  
A. T. Palasz ◽  
H. Rodriguez-Martinez ◽  
P. Beltran-Breña ◽  
S. Perez-Garnelo ◽  
M. F. Martinez ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Patterns ◽  
Gene Expression Patterns ◽  
Bovine Embryo ◽  
In Vitro Development ◽  
Vitro Development

173 THE EFFECT OF AMINO ACIDS AND HYALURONAN ON BOVINE EMBRYO IN VITRO DEVELOPMENT AND GENE EXPRESSION PATTERN

2006 ◽  
Vol 18 (2) ◽  
pp. 194 ◽  
Author(s):  
A. T. Palasz ◽  
J. Beltrán Breña ◽  
P. De la Fuente ◽  
M. F. Martinez ◽  
A. Gutiérrez-Adán
Keyword(s):  
Gene Expression ◽  
Amino Acids ◽  
Embryo Development ◽  
Control Group ◽  
Bovine Embryo ◽  
In Vitro Development ◽  
Glut 1 ◽  
Vitro Development ◽  
And Control

We have previously shown that bovine embryos cultured in SOFaa (BME + MEM amino acids) culture medium with hyaluronan (HA) + BSA are of better quality (Guti�rrez-Ad�n et al. 2005 Reprod. Fertil. Dev. 17, 219). Our objective was to examine the effect of essential (BME) or non-essential (MEM) amino acids with or without HA (MAP-5; Bioniche, Inc., Belleville, Ontario, Canada) on bovine embryo in vitro development and mRNA transcription of five developmentally important genes; apoptosis (Bax), growth factor (IGF-II), glucose (Glut-1) and fructose (Glut-5) transport and metabolism, and cell to cell adhesion (Cx-43). A total of 1474 presumptive zygotes (5 replicates) were initially cultured in 40 �L drops in the following groups: Group 1, control, SOFaa; Group 2, SOF-1 (MEM only); and Group 3, SOF-2 (BME only). On Day 4 (~96 h post-insemination (pi) the number of zygotes that had developed to d8 cells was recorded and 10 �L of SOF-1 and SOF-2, each with 2.5 mg/mL HA, was added to half of the embryos from Groups 2 and 3, respectively; the other half of Groups 2 and 3 and control group received 10 �L of corresponding medium without HA. Embryos were cultured under paraffin oil at 39�C and 5% CO2 in humidified air. Cleavage rates were recorded on Day 2 and the number of blastocysts on Days 7, 8, and 9. Five blastocysts from each replicate from each treatment were frozen for determination of gene expression patterns later. Cleavage rates and embryo development 96 h pi were compared among groups by chi-square analysis. The effects of HA and medium on blastocyst rates were analyzed by logistic regression and the data on mRNA expression by one-way repeated-measures ANOVA. Cleavage rates were 81.1% in SOFaa and 79.3% in SOF-1 (P = 0.48) and different from those in the SOF-2 group (72.4%; P < 0.02). The proportion of embryos that developed to d8 cells at Day 4 was higher in the control (46.7%) and SOF-1 (46.8%) groups than in the SOF-2 group (32.6%). The number of blastocysts that developed in SOFaa (37.0%), SOF-1 (37.7%), and SOF-1 + HA (37.8%) were higher (P < 0.001) than those in SOF-2 (19.6%) and SOF-2 + HA (21.8%). The level of expression of Glut-5 was not different among the groups. However, SOF-2 was the only group that had significantly lower expression of Glut-5, Igf II, and Cx43, and higher expression of BAX (P < 0.05) as compared to the control group and the SOF-1 groups with or without HA. Addition of HA to SOF-2 medium increased expression of Glut-1 and Igf II and decreased expression of BAX as compared to the SOF-1 only and control groups and the SOF-2 groups with or without HA (P < 0.05). The level of expression of Cx43 was higher in the control than in four remaining groups, and lower in the SOF-2 than in the SOF-1 group (P < 0.05). Addition of HA increased expression of Cx43 in both SOF-1 and SOF-2 groups but this level of expression was lower than in the control group; the level in the SOF-2 + HA group was lower (P < 0.05) than in the SOF-1 + HA group. We conclude that, within our protocol, MEM amino acids only stimulate embryo development to the blastocyst stage and the addition of HA to the SOF-MEM and SOF-BME media on Day 4 of culture improved embryo quality.

Gene expression profiles and in vitro development following vitrification of pronuclear and 8-cell stage mouse embryos

2006 ◽  
Vol 73 (6) ◽  
pp. 700-708 ◽  
Author(s):  
Duangjai Boonkusol ◽  
Arpad Baji Gal ◽  
Szilard Bodo ◽  
Botond Gorhony ◽  
Yindee Kitiyanant ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Mouse Embryos ◽  
Cell Stage ◽  
In Vitro Development ◽  
Vitro Development

198 THE EFFECT OF SOURCE AND IN VITRO MATURATION ON THE ABUNDANCE OF MATERNAL mRNA OF SELECTED GENES IN FOLLICULAR BOVINE OOCYTES AND THEIR INFLUENCE ON IN VITRO DEVELOPMENT

2011 ◽  
Vol 23 (1) ◽  
pp. 199
Author(s):  
T. Somfai ◽  
K. Imai ◽  
M. Kaneda ◽  
S. Akagi ◽  
S. Haraguchi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Embryo Development ◽  
In Vitro Maturation ◽  
Blastocyst Stage ◽  
Cell Stage ◽  
In Vitro Development ◽  
Before And After ◽  
Vitro Development ◽  
Bovine Oocytes

The aim of the present study was to investigate the effect of oocyte source and in vitro maturation (IVM) on the expression of selected genes in bovine oocytes and their contribution to in vitro embryo development. Follicular oocytes were collected either by ovum pick-up from live cows or by the aspiration of ovaries of slaughtered cows following storage in Dulbecco’s PBS at 15°C for overnight. In vitro maturation was performed according to the method of (Imai et al. 2006 J. Reprod. Dev. 52, 19–29 suppl.). Gene expression was assessed before and after IVM by real-time PCR. The following genes were investigated: GAPDH, G6PDH, ACTB, H2A, CCNB1, MnSOD, OCT4, SOX2, CX43, HSP70, GLUT8, PAP, GDF9, COX1, ATP1A1, CDH1, CTNNB1, AQP3, DYNLL1, DYNC 1/1, and PMSB1. In brief, mRNA was extracted from 20 oocytes per sample using a Qiagen RNeasy Micro Kit (Qiagen, Valencia, CA). Gene expression was analysed by a Roche Light Cycler 480 device and software (Roche, Indianapolis, IN). Relative expression of each gene was normalized to CCNB1, which in preliminary experiments appeared the most stably expressed irrespective of oocyte source and meiotic stage. Three replications were performed. Data were analysed by paired t-test. In immature ovum pick-up oocytes, genes related to metabolism (GAPDH, G6PDH, GLUT8) and stress (MnSOD, HSP70), and also OCT4, ATP1A1, and DYNC1/1 showed significantly (P < 0.05) higher expression compared with immature oocytes collected from slaughtered-stored ovaries. The expression of GDF9, GLUT8, CTNNB1, and PMSB1 was significantly (P < 0.05) reduced during IVM irrespective of the oocyte source. In a second experiment, IVF IVM oocytes showing an early (at 22 to 25 h after IVF) or late (at 27 to 30 h after IVF) first cleavage were either cultured in vitro or analysed for gene expression at the 2-cell stage. A higher (P < 0.05) rate of early-cleaving oocytes developed to the blastocyst stage compared with the rate of late-cleaving ones (46.2% v. 15.6%, respectively). Nevertheless, only ATP1A1 showed significantly reduced (P < 0.05) expression in late-cleaving embryos compared with early-cleaving ones. Our results suggest that although removal and storage of ovaries and IVM caused a reduction in the relative abundance of several genes in oocytes, in most cases, this did not affect embryo development. Among the genes studied, only ATP1A1 was correlated with in vitro development.

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