Thermotolerance and Antioxidative Capacity of Ear Fibroblasts Derived from Cloned Cattle with Different Ooplasm Origins

• Main Authors: Piyawit Kesorn, 宋亞威
• Other Authors: Perng-Chih Shen
• Format: Others
• Language: en_US
• Published: 2018
• Online Access: http://ndltd.ncl.edu.tw/handle/y4243p

博士 === 國立屏東科技大學 === 熱帶農業暨國際合作系 === 106 === We have previously produced cloned Holstein by using somatic cell nuclear transfer (SCNT) from reconstructed embryos containing ooplasm from different breeds. Results demonstrated that the ear fibroblasts derived from cloned Holstein reconstructed using the Taiwan native yellow cattle ooplasm (Yo) are more thermotolerant than ear fibroblasts derived from cloned Holstein reconstructed using the Holstein ooplasm (Ho). However, molecules mediating the thermotolerance and antioxidant ability of ear fibroblasts of these cloned Holstein remained unclear. Moreover, whether these traits are transmissible to their offspring requires further investigations. The objective of this study was to compare the thermotolerances and antioxidant abilities of ear fibroblasts derived from Holstein (H) and Taiwan native yellow cattle (Y), and cloned Holstein reconstructed with recipient ooplasm from different bovine breeds (Yo-Hd, Ho-Hd), and their offspring (Yo-Hd-F1, Ho-Hd-F1). Polymorphisms in mitochondrial DNA (mtDNA) D-loop of ear fibroblasts derived from Yo-Hd and Yo-Hd-F1 indicated that the cytoplasm is originated from Bos indicus (Y). After heat shock, the apoptotic rates, B-cell lymphoma 2-associated X protein/B-cell lymphoma 2 ratios (Bax/Bcl-2), and relative expression levels of cysteine-aspartic proteases (caspases)-3, -8, and -9 of ear fibroblasts with Y-originated cytoplasm (including Y, Yo-Hd, and Yo-Hd-F1) were all significantly (P < 0.05) lower than those of ear fibroblasts with H-originated cytoplasm (including H, Ho-Hd, and Ho-Hd-F1). In contrast, the relative expression level of HSP-70 was significantly (P < 0.05) higher in ear fibroblasts with Y-originated cytoplasm than that of ear fibroblasts with H-originated cytoplasm. Moreover, oxidative phosphorylation proteins (Complex III and IV) of ear fibroblasts with Y-originated cytoplasm were significantly (P < 0.05) greater than those of ear fibroblasts with H-originated cytoplasm. The catalase (CAT) expressions in heat-shocked ear fibroblasts derived from Yo-Hd and Yo-Hd-F1 cattle were significantly (P < 0.05) higher than those of fibroblasts derived from Ho-Hd-F1 cattle. The level of glutathione peroxidase (GPx) expressions were higher (P < 0.05) in ear fibroblasts with Y-originated cytoplasm compared to those of fibroblasts with H-originated cytoplasm. Furthermore, after exposure to H2O2, the apoptotic rate of ear fibroblasts with Y-originated cytoplasm was lower (P < 0.05) than that of cells with H-originated cytoplasm. The expression of glutathione peroxidase was higher (P < 0.05) in ear fibroblast cells with Y-originated cytoplasm compared to ear fibroblasts with H-originated cytoplasm. In addition, the expression of catalase in ear fibroblasts derived from Yo-Hd and Yo-Hd-F1 cattle was higher, but not statistically significant (P > 0.05) than that of cells derived from Ho-Hd and Ho-Hd-F1 cattle. In conclusion, the ear fibroblasts derived from cloned Holstein reconstructed using the Y ooplasm and Yo-Hd-F1 had better thermotolerances and antioxidant abilities than ear fibroblasts derived from cloned Holstein reconstructed with the H ooplasm and Ho-Hd-F1.