Comparison of activation ability between feline and bovine oocytes

2013 ◽  
Vol 61 (4) ◽  
pp. 491-494 ◽  
Author(s):  
Fuminori Tanihara ◽  
Yukine Kaedei ◽  
Zhao Namula ◽  
Vien Luu ◽  
Yoko Sato ◽  
...  
Keyword(s):  
Chemical Treatment ◽  
Mammalian Species ◽  
Parthenogenetic Activation ◽  
Activation Rates ◽  
Bovine Oocytes ◽  
Higher Sensitivity

Research comparing the activation sensitivity of oocytes to chemical treatment among mammalian species remains limited. We compared the activation ability of oocytes from bovine and feline ovaries when treated with ethanol alone, with ethanol and cycloheximide, and without any chemical treatment; the oocytes were then cultured for 72 h. After in vitro maturation (IVM), 5% of feline oocytes were activated and 1% were cleaved, whereas there were no prematurely activated bovine oocytes. Activation rates with ethanol and ethanol/cycloheximide were significantly higher (P < 0.01) in bovine oocytes than in feline oocytes (74.2% vs. 34.1% and 86.3% vs. 52.5%, respectively). Thus, our findings indicate that feline oocytes can be prematurely activated by the end of IVM, and that bovine oocytes may have a higher sensitivity of parthenogenetic activation to chemical treatment than do feline oocytes.

scholarly journals Oocyte activation and preimplantation development of bovine embryos obtained by specific inhibition of cyclin-dependent kinases

2007 ◽  
Vol 59 (2) ◽  
pp. 280-287 ◽  
Author(s):  
F. Perecin ◽  
S.C. Méo ◽  
C.L.V. Leal ◽  
J.M. Garcia
Keyword(s):  
Embryonic Development ◽  
Developmental Competence ◽  
Oocyte Activation ◽  
Blastocyst Development ◽  
Parthenogenetic Activation ◽  
Cyclin Dependent Kinases ◽  
Activation Rate ◽  
Activation Rates ◽  
Bovine Oocytes

The efficiency of bohemine and roscovitine in combination with ionomycin on parthenogenetic activation and initial embryonic development of bovine oocytes was studied. Two experiments were performed: in the first, different concentrations (0, 50, 75 or 100µM) and different exposure periods (2, 4 or 6 hours) to bohemine or roscovitine were tested for activation rates of in vitro matured (IVM) bovine oocytes, which were pre-exposed to ionomycin. The best treatments, 75µM bohemine and 50µM roscovitine, both for 6h, were used in the second experiment, in which IVM bovine oocytes were exposed to ionomycin, followed or not by bohemine or roscovitine treatment, and evaluated for nuclear status, activation rate and blastocyst development were assessed. The combined treatments (ionomycin + cyclin-dependent kinases inhibitors - CDKIs) showed better results for activation rates (77.3%) and initial embryonic development (35.2%) than the single ionomycin treatment (69.4% for activation and 21.9% for development); and also lead to a more uniform activation (nearly 90% single pronucleus development). The results showed that CDKIs improve the effects of ionomycin on parthenogenetic activation and blastocyst development in bovine oocytes and could help to achieve more efficient activation protocols, increasing the developmental competence of embryos obtained by reproductive biotechniques.

Artificial activation of bovine and equine oocytes with cycloheximide, roscovitine, strontium, or 6-dimethylaminopurine in low or high calcium concentrations

Zygote ◽  
2013 ◽  
Vol 22 (3) ◽  
pp. 387-394 ◽  
Author(s):  
Claudia Barbosa Fernandes ◽  
Liani Gasparini Devito ◽  
Lilian Rigatto Martins ◽  
Ieda Dala Pria Blanco ◽  
João Ferreria de Lima Neto ◽  
...  
Keyword(s):  
Bos Taurus ◽  
Calcium Ionophore ◽  
Parthenogenetic Activation ◽  
High Calcium ◽  
Inverted Microscope ◽  
High Concentrations ◽  
Activation Rates ◽  
Artificial Activation ◽  
Bovine Oocytes

SummaryKnowledge on parthenogenetic activation of oocytes is important to improve the efficiency of nuclear transfer (NT) and intracytoplasmic sperm injection (ICSI) because artificial activation of oocyte (AOA) is an essential step to achieve embryo production. Although different procedures for AOA have been established, the efficiency of in vitro production of embryos remains low, especially in equines and Bos taurus bovines. In an attempt to improve the techniques of NT and ICSI in bovine and equine species, we tested different combinations of drugs that had different mechanisms of action for the parthenogenetic activation of oocytes in these animals. The oocytes were collected, in vitro matured for 24 to 30 h and activated artificially, in the presence of low or high concentrations of calcium, with combinations of calcium ionophore (ionomycin) with cycloheximide, roscovitine, strontium, or 6-dimethylaminopurine (6-DMAP). For assessment of activation rates, oocytes were stained with Hoechst 33342 and observed under an inverted microscope. We showed that all combinations of drugs were equally efficient in activating bovine oocytes, with the best results obtained when high concentrations of calcium were adopted. For equine oocytes, high concentrations of calcium were not beneficial for the parthenogenetic activation and the combination of ionomycin with either 6-DMAP or roscovitine was effective in inducing artificial activation of oocytes. We believe that our preliminary findings provide some clues for the development of a better AOA protocol to be used with these species.

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.

164 EFFECTS OF CUMULUS CELLS AND PITUITARY HORMONES ON AGE-ASSOCIATED CELLULAR CHANGES DURING THE PROLONGED CULTURE OF BOVINE OOCYTES

2014 ◽  
Vol 26 (1) ◽  
pp. 196
Author(s):  
I. Lebedeva ◽  
G. Singina ◽  
A. Lopukhov ◽  
N. Zinovieva
Keyword(s):  
Fetal Calf Serum ◽  
Calf Serum ◽  
Cumulus Cells ◽  
Pituitary Hormones ◽  
Meiotic Arrest ◽  
Parthenogenetic Activation ◽  
Cellular Changes ◽  
Prolonged Culture ◽  
Bovine Oocytes

In vivo and in vitro aging of mature mammalian oocytes heavily reduces their quality and developmental capacity. Therefore, the knowledge of physiological factors modulating the speed of oocyte aging is of great importance for successful reproduction. The goal of the present research was to study effects of cumulus cells (CC) and two related pituitary hormones, prolactin (PRL) and growth hormone (GH), on the dynamics of age-associated cellular changes during the prolonged culture of bovine oocytes in vitro. Bovine cumulus–oocyte complexes (COC) were cultured for 20 h in the following maturation medium: TCM 199 containing 10% fetal calf serum, 10 μg mL–1 porcine FSH, and 5 μg mL–1 ovine LH. After IVM, COC were transferred to the aging medium consisting of TCM-199 supplemented with 10% fetal calf serum and cultured for 0, 12, 24, or 36 h in the absence (Control) or presence of 50 ng mL–1 bovine PRL (Research Center for Endocrinology, Moscow, Russia) or 10 ng mL–1 recombinant bovine GH (Monsanto, St. Louis, MO, USA). A portion of in vitro-matured oocytes were denuded of their CC and cultured in the control aging medium. At the end of culture, the state of the nuclear material in oocytes and embryos was evaluated by Tarkowski's cytogenetic method. The number of oocytes undergoing spontaneous parthenogenetic activation in the respective groups was determined by summarising the numbers of embryos cleaved and oocytes reaching anaphase-II to telophase-II stages or containing a pronucleus. Destructive changes in CC were assessed using the morphological signs of apoptosis. The data from 3 to 5 replicates were analysed by ANOVA. In the control group of COC, a rise in the rate of metaphase-II (M-II) oocytes with abnormal chromosome configurations occurred by 12 h of aging [31.8 ± 4.6% (12 h) v. 17.5 ± 2.6% (0 h); P < 0.05] and persisted up to 36 h (70.4 ± 2.0%; P < 0.001). At the same time, the frequency of oocyte parthenogenetic activation markedly increased only between 0 and 36 h of aging (from 0% to 20.7 ± 3.4%; P < 0.001). The addition of PRL or GH to the aging medium or removal of CCs resulted in a decline in the rate of M-II oocytes with degenerative changes of chromosomes throughout the culture period (at least P < 0.05). Furthermore, PRL and GH reduced the frequency of the oocyte activation at 36 h of the prolonged culture (up to 5.4 ± 2.5 and 1.7 ± 1.7%, respectively; P < 0.01), although CC did not influence meiotic arrest at M-II. Meanwhile, the rate of degenerated CC steadily increased as the culture time increased from 0 h (10.3 ± 1.1%) to 36 h (22.7 ± 2.2%; P < 0.001) and was unaffected by both hormones. The data suggest that, in bovine COC, CC accelerate abnormal changes in the chromosomal structure of aging M-II oocytes, whereas PRL and GH may decelerate these changes and support meiotic arrest during the prolonged culture of in vitro-matured oocytes. This research was partially supported by RFBR (project no. 13-04-01888).

In vitro and in vivo survival of frozen-thawed bovine oocytes after IVF, nuclear transfer, and parthenogenetic activation

1998 ◽  
Vol 51 (3) ◽  
pp. 281-286 ◽  
Author(s):  
Chikara Kubota ◽  
Xiangzhong Yang ◽  
Andras Dinnyes ◽  
Junichi Todoroki ◽  
Hiroshi Yamakuchi ◽  
...  
Keyword(s):  
Nuclear Transfer ◽  
Parthenogenetic Activation ◽  
Bovine Oocytes

Protein phosphorylation in bovine oocytes following fertilization and parthenogenetic activation in vitro

1997 ◽  
Vol 47 (1) ◽  
pp. 206
Author(s):  
H.M. Chung ◽  
R.C. Chian ◽  
J.J. Ko ◽  
T.K. Yoon ◽  
K.Y. Cha
Keyword(s):  
Protein Phosphorylation ◽  
Parthenogenetic Activation ◽  
Bovine Oocytes

Optimized combined electrical–chemical parthenogenetic activation for in vitro matured bovine oocytes

2008 ◽  
Vol 108 (1-2) ◽  
pp. 122-133 ◽  
Author(s):  
S.M. Hosseini ◽  
M. Hajian ◽  
F. Moulavi ◽  
A.H. Shahverdi ◽  
M.H. Nasr-Esfahani
Keyword(s):  
Parthenogenetic Activation ◽  
Bovine Oocytes
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