Cellular and molecular alterations of buffalo oocytes cultured under two different levels of oxygen tension during <i>in vitro</i> maturation

Ghanem, Nasser; Ahmed, Dalia Abd-El Rahman; Dessouki, Sherif Mohamed; Faheem, Marwa; Gad, Ahmed; Peippo, Jaana; Barkawi, Ashraf Hesham

doi:10.1017/s0967199420000945

Cited by 4 publications

(2 citation statements)

References 70 publications

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“…The reaction mix was incubated at 50°C for 2 minutes; initial denaturation was done for 10 minutes at 95°C and 40 cycles at 95°C (Denaturation) for 15 minute and final step at 60°C for 1 minute (annealing), at 95°C for 15 seconds and then 60°C for 1 minute. The results were expressed as Ct values and relative gene expression profile was estimated delta delta Ct analysis ( 2-ΔΔCT method) as done in our recent study (Ghanem et al, 2021).…”

Section: Quantitative Real-time Pcrmentioning

confidence: 99%

Co-expression of candidate genes regulating growth performance and carcass traits of Barki lambs in Egypt

Ghanem¹,

Zayed²,

Mohamed³

et al. 2021

Preprint

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The aim of present study was to link the gene expression profile of selected candidate genes with blood profile, growth performance and carcass traits of Barki lambs. Thirty-eight Barki lambs were divided into 3 groups (fast, intermediate and slow growing) according to growth perfor-mance. Body tissues (muscle, liver and fat) were taken from for RNA isolation and Real-time PCR. The results indicated that, the final body weight hot carcass weight were heavier (P ≤ 0.05) in fast (49.9 Kg and 24.57) than intermediate (40.7 and 19.07 Kg) and slow (30.8 and 15.10 Kg) growing animals. The blood profile of total protein, total lipids, calcium, T3 and T4 hormones did not differ among sheep groups. Genes involved in protein biosynthesis (RPL7), fatty acid oxidation (CPT1) and lipolysis (FABP4) were up regulated in fast and intermediate growing lambs in all studied tissues. While, gene-regulating lipogenesis (ADIPOQ) was expressed simi-larly in fat and liver tissues, but increased its expression in muscle of fast and intermediate growing lambs. Expression of CAPN3 was increased in fast and intermediate growing compared to slow growing lambs. In conclusion, the current study providing an evidence for the im-portance of co-expression of these genes in main body tissues linked with growth performance of Barki lambs.

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Section: Quantitative Real-time Pcrmentioning

confidence: 99%

Co-expression of candidate genes regulating growth performance and carcass traits of Barki lambs in Egypt

Ghanem¹,

Zayed²,

Mohamed³

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…The reaction mix was incubated at 50°C for 2 minutes; initial denaturation was done for 10 minutes at 95°C and 40 cycles at 95°C (Denaturation) for 15 minute and nal step at 60°C for 1 minute (annealing), at 95°C for 15 seconds and then 60°C for 1 minute. The results were expressed as Ct values and relative gene expression pro le was estimated delta delta Ct analysis (2-ΔΔCT method) as done in our recent study (Ghanem et al, 2021).…”

Section: Quantitative Real-time Pcrmentioning

confidence: 99%

Co-Expression of Candidate Genes Regulating Growth Performance and Carcass Traits of Barki Lambs in Egypt

Ghanem

Zayed

Mohamed

et al. 2021

Preprint

Self Cite

View full text Add to dashboard Cite

Sheep are considered one of the main sources of animal protein in Egypt while surviving harsh desert conditions. Therefore, the aim of present study was to link the gene expression profile of selected candidate genes with growth performance and carcass traits of Barki lambs. Thirty-eight Barki lambs were kept and fed individually after weaning till 12 months of age. Lambs were divided into 3 groups (fast, intermediate and slow growing) according to growth performance. Carcass traits were recorded directly after slaughtering. Three samples were taken from different body tissues (eye muscle, liver and fat tail) of each group, directly during slaughtering and stored at -80°C until RNA isolation. Real-time PCR was used to profile selected candidate genes (RPL7, CTP1, FABP4, ADIPOQ and CAPN3) and GAPDH was used as a housekeeping gene. The results indicated that, the final body weight was significantly (P ≤ 0.05) greater in fast (49.9 Kg) and intermediate (40.7 Kg) compared to slow growing animals (30.8 Kg). The hot carcass weight was heavier (P ≤ 0.05) in fast, intermediate (24.57 and 19.07 Kg) than low growing lambs (15.10 Kg). The blood profiles of T3 and T4 hormones showed no significant different among fast (1.1 ng/ml and 8.5 µg/dl), intermediate (0.6 ng/ml and 12.4 µg/dl) and slow growing lambs (1.0 ng/ml and 10.5 µg/dl). In addition, the total protein level was not differ in fast (5.94 g/dl), intermediate (6.52 g/dl) and slow growing lambs (6.9 g/dl). Genes involved in protein biosynthesis (RPL7), fatty acid oxidation (CPT1) and lipolysis (FABP4) were up regulated in fast and intermediate growing lambs in all studied tissues. While, gene-regulating lipogenesis (ADIPOQ) was expressed similarly in fat and liver tissues, but increased its expression in muscle of fast and intermediate growing lambs. Expression of CAPN3 was increased in fast and intermediate growing compared to slow growing lambs. In conclusion, the current study indicated a link between variation in gene expression profile in main body tissues with growth performance and carcass weight of Barki lambs, which induced by the genetic makeup however; this hypothesis required a large population for validation. In addition, the transcriptional profile of CPT1, FABP4 RPL7 and CAPN3 are linked with growth performance and carcass traits of Barki lambs, providing an evidence for the importance of co-expression of these genes in body main tissues involved in this process.

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CX43 and oxidative stress are the targets of BCB staining to predict the developmental potential of buffalo oocytes

Li,

Yang,

Duan

et al. 2024

Reprod Domestic Animals

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This study used the brilliant cresyl blue (BCB) staining method to group buffalo oocytes (BCB+ and BCB−) and perform in vitro maturation, in vitro fertilization and embryo culture. At the same time, molecular biology techniques were used to detect gap junction protein expression and oxidative stress‐related indicators to explore the molecular mechanism of BCB staining to predict oocyte developmental potential. The techniques of buffalo oocytes to analyse their developmental potential and used immunofluorescence staining to detect the expression level of CX43 protein, DCFH‐DA probe staining to detect ROS levels and qPCR to detect the expression levels of the antioxidant‐related genes SOD2 and GPX1. Our results showed that the in vitro maturation rate, embryo cleavage rate and blastocyst rate of buffalo oocytes in the BCB+ group were significantly higher than those in the BCB− group and the control group (p < .05). The expression level of CX43 protein in the BCB+ group was higher than that in the BCB− group both before and after maturation (p < .05). The intensity of ROS in the BCB+ group was significantly lower than that in the BCB− group (p < .05), and the expression levels of the antioxidant‐related genes SOD2 and GPX1 in the BCB+ group were significantly higher than those in the BCB− group (p < .05). Brilliant cresyl blue staining could effectively predict the developmental potential of buffalo oocytes. The results of BCB staining were positively correlated with the expression of gap junction protein and antioxidant‐related genes and negatively correlated with the reactive oxygen species level, suggesting that the mechanism of BCB staining in predicting the developmental potential of buffalo oocytes might be closely related to antioxidant activity.

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Cellular and molecular alterations of buffalo oocytes cultured under two different levels of oxygen tension during in vitro maturation

Cited by 4 publications

References 70 publications

Co-expression of candidate genes regulating growth performance and carcass traits of Barki lambs in Egypt

Co-expression of candidate genes regulating growth performance and carcass traits of Barki lambs in Egypt

Co-Expression of Candidate Genes Regulating Growth Performance and Carcass Traits of Barki Lambs in Egypt

CX43 and oxidative stress are the targets of BCB staining to predict the developmental potential of buffalo oocytes

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