Electrocatalytic oxidation of NADH by Brilliant Cresyl Blue–DNA intercalation adduct

2005 ◽  
Vol 50 (5) ◽  
pp. 1107-1112 ◽  
Author(s):  
Patricia de-los-Santos-Álvarez ◽  
M. Jesús Lobo-Castañón ◽  
Arturo J. Miranda-Ordieres ◽  
Paulino Tuñón-Blanco
2003 ◽  
Vol 43 (2) ◽  
pp. 179-187 ◽  
Author(s):  
Elisabeth Rodríguez-González ◽  
Manel López-Bejar ◽  
Dolors Izquierdo ◽  
María-Teresa Paramio

2021 ◽  
Vol 14 (2) ◽  
pp. 452-456
Author(s):  
Mohamed Fathi ◽  
Amr F. Elkarmoty

Aim: Several factors had been concerned with the developmental competence of the sheep oocyte. This study aims to investigate the effect of adding growth factors (insulin-like growth factor 1 [IGF-1] and epidermal growth factor [EGF]) in the maturation medium of ewe oocytes selected based on brilliant cresyl blue (BCB) screening on in vitro maturation (IVM), fertilization, and pre-implantation embryo development. Materials and Methods: Cumulus-oocyte complexes (COCs) were obtained from the ovaries of slaughtered ewes by either aspiration or slicing techniques. COCs were in vitro matured in a medium containing IGF-1 and EGF (control group). For BCB screening, oocytes were stained and divided into BCB+ oocytes that matured in the same maturation conditions without adding growth factors (Group 2) or in the presence of growth factors (Group 3), and BCB– oocytes that matured in medium without growth factors (Group 4) or with growth factors (Group 5). Results: The supplementation of the maturation medium with growth factors during IVM of (BCB+) oocytes resulted in a significant increase in nuclear maturation rate (90.9%), fertilization rate (75.6%), and embryo developmental rates (60.0%, 46.7%, and 33.3% for cleavage, morula, and blastocyst, respectively). Conclusion: Culturing BCB+ oocytes in a maturation medium containing both EGF and IGF-1 showed a significant improvement in nuclear maturation, fertilization, and pre-implantation embryo development in vitro.


2013 ◽  
Vol 136 (4) ◽  
pp. 245-251 ◽  
Author(s):  
S.M. Mirshamsi ◽  
H. KaramiShabankareh ◽  
M. Ahmadi-Hamedani ◽  
L. Soltani ◽  
H. Hajarian ◽  
...  

Nature ◽  
1965 ◽  
Vol 207 (4994) ◽  
pp. 329-329 ◽  
Author(s):  
B. C. ARNOLD

1969 ◽  
Vol 17 (2) ◽  
pp. 138-144 ◽  
Author(s):  
R. D. Lillie ◽  
P. Pizzolato

2013 ◽  
Vol 25 (1) ◽  
pp. 246
Author(s):  
M. G. Catalá ◽  
M. Roura ◽  
D. Izquierdo ◽  
S. Hammammi ◽  
S. Uzbekova ◽  
...  

Brilliant cresyl blue (BCB) staining determines the activity of glucose-6-phosphate dehydrogenase (G6PDH), an enzyme in which activity decreases as the oocytes reach their growth phase. We have previously shown in lamb the effectiveness of this stain in selecting the largest and most competent oocytes to develop up to the blastocyst stage (Catala et al. 2011 Reproduction 142, 517–527). The aim of this study was to analyse the expression of 4 genes related to oocyte quality in BCB-selected oocytes. The 4 genes analyzed in this study were selected after a literature review in which they were associated with a better oocyte quality or embryo development; 2 of these genes were related to cell cycle: nuclear auto-antigenic sperm protein (NASP) and histone H2A (H2A.Z), and 2 genes with enzymatic functions: peroxiredoxin (PRDX1) and elongation factor-A1 (EEF1A1). Oocytes were recovered after slicing the surface of lamb ovaries (3 to 5 months old) obtained from a local slaughterhouse. Oocytes with more than 3 compact cumulus layers and homogenic cytoplasm were selected and exposed to 13 µM BCB during 1 h before IVM and classified according to oocyte coloration: oocytes with blue cytoplasm or grown oocytes (BCB+) and oocytes not coloured or growing oocytes (BCB–). Cumulus–oocyte complexes were then matured in conventional TCM199 medium supplemented with 10% fetal bovine serum and hormones during 24 h in a controlled atmosphere. Groups of 15 denuded oocytes (3 replicates) were taken before (0 h) and after maturation (24 h) and stored at –80°C in 100 mL of Trizol until use. After RNA extraction following manufacturer’s protocol (Promega, Madison, WI, USA), reverse transcription was performed by extended cDNA using Oligo (dT) primers during 5 min at 70°C and 1 h at 65°C using superscript III (200 U mM–1; Invitrogen). Relative qualitative PCR analyses were performed in duplicate using SYBR Green Fluorophore kit (Bio-Rad, Hercules, CA, USA). The specificity of each PCR product was determined by a melting curve analysis and the amplicon size determination by an agarose gel. A standard curve was also included, consisting of corresponding plasmid DNA fragments from 1 pg to 0.1 fg, purified with QIAquick PCR Purification Kit (Qiagen, Valencia, CA, USA). Correlation coefficients and PCR efficiencies were considered between 85 and 100%. Finally, the results for mRNA were normalized according to the relative concentration of the internal and external gene (luciferase and 18S, respectively). The statistical analysis was performed by One-way ANOVA in GraphPad Prism v 3 (GraphPad Software, San Diego, CA, USA). The analysis of gene expression showed no differences in relation to BCB classification. The only difference we found is a significant decrease (P ≤ 0.05) in the RE of PRDX observed in matured oocytes compared with immature ones. In conclusion, the expression of the mRNA expression of NASP, H2A.Z, PRDX1, and EEF1A1 were not affected by oocyte quality.


2015 ◽  
Vol 27 (1) ◽  
pp. 235
Author(s):  
K. P. M. Lekola ◽  
J. W. Ng'ambi ◽  
N. Nkadimeng ◽  
M. L. Mphaphathi ◽  
T. L. Nedambale

In vitro maturation of indigenous African cattle oocytes is a major challenge even though different maturation protocols work successfully in other breeds. The objective of this study was to determine the maturation rate of indigenous South African cattle oocytes following in vitro maturation in media supplemented with different concentrations of hormones and selected using brilliant cresyl blue (BCB) staining. Indigenous cattle ovaries were collected from the slaughterhouse and then oocytes were retrieved by aspiration method. A total of 966 oocytes were exposed to 26 µM BCB stain and 700 oocytes were not exposed to the BCB stain. Thereafter, oocytes exposed to the BCB stain were grouped according to the colour of their cytoplasm BCB+ (oocytes with blue cytoplasm, low G6PDH) and BCB– (unstained oocytes, increased G6PDH). The BCB exposed (BCB+ and BCB–) and the oocytes not exposed to BCB were then randomly allocated into tissue culture medium (TCM199) + 10% (vol/vol) fetal bovine serum (FBS) supplemented with 3 different concentrations of hormones as treatments (T). The T1 group was matured in the presence of 0.5 µg mL–1 of FSH, 5 mg mL–1 of LH, and 2 µg mL–1 of E2; the T2 group was matured in the presence of 1 µg mL–1 of FSH, 6 mg mL–1 of LH, and 2.5 µg mL–1 of E2; and the T3 group was matured in the presence of 1.5 µg mL–1 of FSH, 7 mg mL–1 of LH, and 4.5 µg mL–1 of E2. For IVM, 20 to 25 COC were placed in 50-µL droplets of IVM medium containing the 3 different levels of hormones. Maturation rate of oocytes was determined by the extrusion of the first polar body after 24 h of incubation in maturation medium. Data was analysed by ANOVA using SAS with 4 replicates per treatment. Treatment 2 yielded higher maturation rate for both BCB+ (65.6%) and not exposed to BCB (60.3%) oocytes compared to T1 (22, 3.03, and 16% for BCB+, BCB–, and not exposed to BCB, respectively) and T3 (48, 2.2, and 48% for BCB+, BCB–, and not exposed to BCB respectively). However, BCB– oocytes had lower polar body extrusion for T1, T2, and T3 (3.03, 8.1, and 2.2%, respectively) compared to BCB+ oocytes (22, 65.6, and 48% for T1, T2, and T3, respectively). In conclusion, immature oocytes that were cultured into TCM199 supplemented with 10% FBS, 1 µg mL–1 of FSH, 6 mg mL–1 of LH, and 2.5 µg mL–1 of E2 showed maturation rate for BCB+ oocytes and those not exposed to BCB. Oocytes selection using BCB staining was a useful test to classify good quality cattle oocytes. Therefore, it is suggested that treatment 2 is a suitable in vitro-maturation medium to mature indigenous South African cattle oocytes.


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