Paternal influence on the time of first embryonic cleavage post insemination and the implications for subsequent bovine embryo development in vitro and fertility in vivo

2001 ◽  
Vol 60 (1) ◽  
pp. 47-55 ◽  
Author(s):  
Fabian Ward ◽  
Dimitrios Rizos ◽  
Doreen Corridan ◽  
Katherine Quinn ◽  
Maurice Boland ◽  
...  
2007 ◽  
Vol 19 (1) ◽  
pp. 262 ◽  
Author(s):  
I. Dimitriadis ◽  
E. A. Rekka ◽  
E. Vainas ◽  
G. S. Amiridis ◽  
C. A. Rekkas

The substrates used in in vitro embryo production (IVP) mimic the in vivo fluids in which oocytes mature, oocytes are fertilized, and the early embryos develop (follicular and oviductal fluid). It is well established that oxidative stress negatively affects in vitro culture (IVC) outcomes. Guaiazulene (G) is a component of chamomile species oil with known antioxidant properties. In the present study, all IVP media were modified by the addition of G solutions so that the former exhibited a total protection against induced lipid peroxidation (TPaLP) similar to that of the respective in vivo environment. The IVP outcomes were then compared between G-processed and control oocytes. Bovine preovulatory follicular (BF) and oviductal (BO) fluid samples were collected from 10 Holstein 4- to 5-year-old cows in estrus. TPaLP was assessed according to the samples' ability to inhibit rat hepatic microsomal lipid peroxidation, by determination of the 2-thiobarbituric acid reactive material. TPaLP (mean % � SEM) of the BF and BO were 70.63 � 10.03 and 16.33 � 4.33, respectively, whereas those of the IVP [in vitro-matured (IVM), in vitro-fertilized (IVF), and IVC] media were lower (17.94 � 1.66, -1.82 � 0.78, and 14.57 � 1.26, respectively). TPaLP of the 0.1 mM G-modified IVP medium increased to 67.2 � 5.85, 19.98 � 2.49, and 69.19 � 6.22, respectively. A total of 2041 class A oocytes were used. The proportion of cleavage, early embryo development (embryos with more than 4 cells), or both after IVP (18 h IVM–5% CO2 in air, and 18 h IVF, 48 h IVC–5% CO2, 10% O2, 85% N) in the presence of G (n = 1237) during each of the IVP phases or any possible combination of IVP phases was compared with the respective control (C, n = 804). Statistical analysis was performed by a chi-squared test; P < 0.05 was considered significant. G improved cleavage and embryo development rates when present during IVM (79.4 and 57.8% vs. 64.5 and 38.2% for C) or both IVM and IVC (78.0 and 60.7% vs. 57.8 and 36.5%, respectively). When present only during 18 h of IVF, G had no effect on embryo production. However, an increased embryo development rate resulted from the combined exposure to G during IVF and IVM (56.4 vs. 29.6%), during IVF and IVC (55.3 vs. 35.5%), or at all IVP phases (56.6 vs. 34.9%). The latter effect resembled the one obtained after G addition only to the IVC medium (62.5 vs. 39.7%, respectively). We concluded that the addition of G to IVP substrates, at concentrations that mimic the in vivo TPaLP conditions, could promote bovine IVP efficiency.


2009 ◽  
Vol 21 (1) ◽  
pp. 104
Author(s):  
J. T. Aaltonen ◽  
K. J. Mattson ◽  
N. M. Loskutoff

As described in the IETS Manual (Stringfellow and Seidel, 1995), and endorsed by the OIE, trypsin can be used (for specific pathogens and livestock) to effectively remove certain infectious agents from in vivo-derived embryos for international transport. Because of the multimillion-dollar AI industry for livestock, the OIE has encouraged more research in developing similar decontamination techniques for semen as an added safeguard to animal quarantine for the prevention of disease transmission. Most or all of the earlier studies on embryos used a porcine pancreatic-derived trypsin. Because of more stringent guidelines from international regulatory agencies on the use of animal products, several serine protease recombinants are now available. Previous experiments comparing the porcine pancreatic extract with a recombinant bovine sequence trypsin developed in corn resulted in no statistical difference in cleavage or morula/blastocyst rates. (Mattson et al. 2008 Theriogenology 69, 724–727). An additional in vivo study treating bovine sperm with a yeast-derived human-sequence trypsin resulted in significantly more transferable-quality embryos after the AI of superovulated cows as compared with sperm not treated with trypsin (Blevins et al. 2008 Reprod. Fertil. Dev. 20, 84). The goal of this experiment was to examine the in vitro development of bovine embryos produced from sperm treated with a recombinant trypsin found in a commercially available density gradient centrifugation (DGC) product (Bovipure, Nidacon, Sweden) compared with DGC without trypsin. Oocyte aspiration, maturation, fertilization, and embryo culture were performed using standard methods in 5 replications (n = 2220 oocytes). Semen was collected and pooled from 2 Bos taurus bulls and frozen in an egg-yolk cryodiluent (Biladyl, Minitube). The semen was processed using Bovipure DGC composed of 2 mL of 40% colloid of silane-coated silica particles containing either a yeast-derived human sequence recombinant trypsin containing no animal by-products (n = 1126 oocytes) or the same colloid without trypsin as the control group (n = 1094 oocytes). Both 40% concentrations were layered over 2 mL of an 80% concentration of the same colloid without any additives. The density gradients were centrifuged at 300g for 20 min, after which time the pellets were washed in 5 mL of prewarmed TL Hepes solution (Cambrex) and centrifuged at 500g for 10 min. The resulting sperm pellets were then resuspended in a volume calculated to provide 1 × 106 sperm mL–1, to be used for in vitro inseminations. Results were compared using a 2-tailed unpaired t-test. Cleavage rates for the trypsin-treated sperm (n = 969, 35.8%) and the control (n = 950, 44.3%) groups were not statistically different (P = 0.20). Although more embryos reached the morula to blastocyst stages in the control group (n = 421, 61.0%) than in the trypsinized group (n = 347, 54.7%), these differences also were not statistically significant (P = 0.85). In conclusion, trypsinized Bovipure DGC of sperm before insemination showed no detrimental effects on IVF-derived bovine embryo development.


Reproduction ◽  
1994 ◽  
Vol 100 (1) ◽  
pp. 131-136 ◽  
Author(s):  
L. A. Johnston ◽  
J. J. Parrish ◽  
R. Monson ◽  
L. Leibfried-Rutledge ◽  
J. L. Susko-Parrish ◽  
...  

1996 ◽  
Vol 8 (5) ◽  
pp. 835 ◽  
Author(s):  
T Pinyopummintr ◽  
BD Bavister

Effects of amino acids on early bovine embryo development in vitro were examined using a chemically-defined, protein-free culture medium. Bovine embryos produced in vitro were cultured from 18 h to 72 h post insemination in a simple medium containing lactate as the only energy source except for the amino acid treatments. Subsequently, embryos were transferred to TCM-199 supplemented with serum for blastocyst development to substantiate their developmental competence. Treatments were: (1) non-essential amino acids from TCM-199 (NEA); (2) essential amino acids from TCM-199 (EA); (3) NEA+EA; (4) Eagle's minimum essential medium amino acids (MEM AA); (5) 11 amino acids present in HECM-6 (11 AA); and (6) 0.2 mM glutamine (GLN). A higher proportion of embryos (percentage of inseminated ova) cleaved to the > or = 8-cell stage by 72 h post insemination in NEA (56.7%), EA (41.2%), 11 AA (40.3%) and GLN (51.1%) than in either NEA+EA (30.0%) or MEM AA (33.1%). However, after transfer to complex medium, embryos that had developed in EA, as well as those in MEM AA or NEA+EA, produced significantly fewer blastocysts (37.1%, 34.4% and 25.6% respectively) than those in NEA (56.7%), GLN (48.9%) or 11 AA (37.7%). The ability of blastocysts to hatch from their zonae pellucidae was also affected by amino acid treatment during cleavage stages. The present study indicated that the addition of NEA or GLN or 11 AA to a chemically-defined culture medium during the cleavage phase of bovine embryo development increases their subsequent ability to reach the blastocyst stage. These data have implications for understanding the nutritional needs of bovine embryos produced in vitro and for optimizing the composition of culture media to support their development.


1986 ◽  
Vol 64 (10) ◽  
pp. 2227-2238 ◽  
Author(s):  
J. H. N. Schel ◽  
H. Kieft

A culture method is described which allows the continuous supply of fresh liquid medium and which prevents the accumulation of toxic metabolites. Development of maize embryos and endosperm after various periods of in vitro ovary culture was studied by light and electron microscopy. Using this method the ultrastructural features of embryo development in vitro were similar to those of in vivo embryos. In contrast, the formation of endosperm was irregular with the absence of cellularization of the inner endosperm being frequent. In some cases, only the endosperm developed without any indication of embryo formation. In a calcium-depleted medium, embryo development was normal but again, endosperm formation was aberrant. No cells were formed in the central part of the endosperm and near the placental region degeneration took place, resulting in vacuoles with dark inclusions, clumps of rough endoplasmic reticulum membranes, and cellular breakdown. The events occurring after in vitro culture strongly resemble those taking place after intergeneric crosses or crosses between diploid and tetraploid strains. It is concluded that defective endosperm development is probably the main factor for the failure of embryo development.


1998 ◽  
Vol 49 (1) ◽  
pp. 173 ◽  
Author(s):  
M. O'Kearney-Flynn ◽  
M. Wade ◽  
P. Duffy ◽  
V. Gath ◽  
M.P. Boland ◽  
...  

2006 ◽  
Vol 18 (2) ◽  
pp. 119
Author(s):  
S. Arat ◽  
H. Bagis ◽  
A. Tas ◽  
T. Akkoc

The activation of oocytes is one of the most important steps for a successful cloning and has great importance on embryo development in vitro. The objective of this study was to examine the different parameters affecting parthenogenetic embryo development in vitro. In the first experiment, two activation protocols were compared to examine the effect of electrical pulse on activation. Bovine oocytes isolated from slaughterhouse ovaries were matured in TCM-199 supplemented with fetal bovine serum (FBS), sodium pyruvate, penicillin/streptomycin, rat insulin-like growth factor (rIGF-1), bovine follicle-stimulating hormone (bFSH), and bovine luteinizing hormone (bLH). A group of oocytes was exposed to a DC pulse of 133 V/500 �m for 25 �s, and then activated by calcium ionophore (5 �M) for 10 min, cytochalasin D (CD) (2.5 �g/mL) + cycloheximide (CHX, 10 �g/mL) for 1 h, and CHX alone for 5 h (Group 1). Another group of oocytes was activated only by chemicals without electrical pulse. Activated oocytes were cultured for 72 h in G1-3 and then 4-6 days in G2-3 medium. In the second experiment, oocytes activated by electrical pulse and chemicals were cultured in Barc medium for 7-9 days or 72 h in G1-3 and then 4-6 days in G2-3 medium. In the third experiment, oocytes activated by electrical pulse and chemicals were cultured for 48 h or 72 h in G1-3 and then 5-7 days or 4-6 days in G2-3 medium. The differences among groups were analyzed by one-way ANOVA after arcsin square transformation. In the first experiment, cleavage rate (75.6%), development rate (37.3%), and blastocyst cell number (78.4 � 3.2) of oocytes activated by electrical pulse was higher than for the group without electrical pulse (28.7%, 8.0%, 59.5 � 4.3, respectively; P < 0.05). This result showed that activation was started more effectively by electrical pulse than by chemicals. In the second experiment, there was no significant difference on cleavage rate between the two groups (66.6%, 65.0%, respectively), and the blastocyst development rate of parthenogenetic embryos cultured in G1-3/G2-3 (36.6%) was higher than in the Barc medium group (16.6%; P < 0.05). This result showed that G1-3/G2-3 medium was more effective for parthenogenetic embryo development than Barc medium. In the third experiment, although significant differences could not be found between the two groups in the development rate of parthenogenetic embryos cultured for a total of 7-9 days (30.8%, 39.2%, respectively), the development rate of embryos cultured for 72 h in G1-3 was higher (26.4%) than for the 48-h group (15%; P < 0.05) on Day 7. This result showed that embryos developed more slowly when cultured for a shorter time in G1-3 medium before transfer to G2-3 medium. This study was supported by a grant from TUBITAK, Turkey (VHAG-1022).


2016 ◽  
Vol 28 (2) ◽  
pp. 173
Author(s):  
I. Khan ◽  
K.-L. Lee ◽  
A.-N. Ha ◽  
P.-R. Park ◽  
S.-H. Song ◽  
...  

Coagulansin-A (withanolide) is the steroidal lactone obtained from Withania coagulans, which belong to Solanaceae family. The coagulansin-A induces heat shock protein 70 (HSP-70), which acts as a cellular antioxidant. This study was conducted to investigate the effect of coagulansin-A on bovine oocytes maturation and embryo development in vitro. All these oocytes were aspirated from the ovaries obtained from Korean Hanwoo cows at a local abattoir. To analyse the possible beneficial effect of coagulansin-A on bovine oocytes maturation in vitro, 355 oocytes per group (control and treatment) in seven replicates were subjected with three concentrations i.e. (1, 5, and 10 µM) of coagulansin-A. The coagulansin-A was added in the in vitro-matured (IVM) media for 20 to 22 h followed by IVF for 18 to 22 h, and after fertilization the fertilized oocytes were transferred to IVC1 media for 3 days. After 3 days, the cleavage rate was checked and the 8-cell stage embryos were transferred to IVC2 media and embryo development was checked at Day 8. The culture was carried out at 5% CO2 and 38.5°C. The results indicated that among the three concentrations of Coagulansin-A, only 5 µM remarkably (P < 0.05) improved embryo development (Day 8 blastocyst), being 27.30% and 40.01% for control and treated groups, respectively. This concentration also significantly (P < 0.05) encouraged the activation of HSP-70, having 16.44 arbitrary units (AU) and 35.41 AU integral optical density (IOD) for control and treated groups, respectively. The immunofluorescence analysis revealed that 5 µM coagulansin-A supplementation significantly (P < 0.05) inhibited oxidative stress and inflammation during bovine embryo development in vitro by decreasing IOD of 8-Oxoguanosine (8-OxoG) from 28.12 AU in control to 18.06 AU for the treated group and nuclear factor kappa B (NF-kB) IOD (P < 0.05) from 42.25 AU to 21.80 for control and treated groups, respectively. Additionally, the results obtained from terminal deoxynucleotidyl transferase TUNEL assay confirmed that coagulansin-A treatment reduced the bovine embryo DNA damage significantly (P < 0.05) from 7.4 ± 0.375 to 5.7 ± 0.287 and improved the embryo quality (P < 0.05) with mean cell numbers of 127.7 ± 4.161 and 150.1 ± 3.624 per embryo for control and coagulansin-A treated groups, respectively. This study provides new information regarding the mechanisms by which coagulansin-A promotes bovine oocyte maturation and embryo development in vitro.


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