旨在为牛体细胞克隆胚胎体外培养体系的建立奠定基础。利用体细胞核移植技术生产牛重构胚,将重构胚随机分成2组,分别用mSOF和G1.5/G2.5培养,统计2种培养液对牛核移植克隆胚胎发育的影响,用TUNEL荧光检测系统检测2个培养组胚胎细胞凋亡情...旨在为牛体细胞克隆胚胎体外培养体系的建立奠定基础。利用体细胞核移植技术生产牛重构胚,将重构胚随机分成2组,分别用mSOF和G1.5/G2.5培养,统计2种培养液对牛核移植克隆胚胎发育的影响,用TUNEL荧光检测系统检测2个培养组胚胎细胞凋亡情况,同时用Real time RT-PCR检测热应激蛋白基因Hsp70和凋亡相关基因Bax在2个培养组的相对表达水平。两组的卵裂率分别为85.6%和87.0%、囊胚率分别为22.9%和27.1%,差异都不显著(P>0.05),但G1.5/G2.5组的8-细胞形成率显著高于mSOF组(P<0.05);mSOF组凋亡率为12.1%±1.9%,显著高于G1.5/G2.5组(P<0.05),G1.5/G2.5组的Hsp70和Bax相对表达量显著低于mSOF组(P<0.05)。结果表明,G1.5/G2.5培养液更有利于牛体细胞核移植胚胎的体外发育。展开更多
Seasonal variations play an essential role in the metabolism, behavior and activity of the laboratory animals. This study was aimed to examine whether mitochondrial function can be influenced by the seasonal changes a...Seasonal variations play an essential role in the metabolism, behavior and activity of the laboratory animals. This study was aimed to examine whether mitochondrial function can be influenced by the seasonal changes and how large is the impact of these fluctuations on experiments with using animal models and further results interpretation. Liver mitochondria were isolated from male Wistar rats and exposed to calcium ions, PAMAM dendrimers G2.5 or their combination: (Ca2+) and dendrimer. The scientific hypothesis assumed that dendrimer G2.5 is able to limit the detrimental effect of Ca2+ ions on mitochondria function, possibly through affecting the following parameters: calcium transport, mitochondrial potential and membrane fluidity. The activity of mitochondria was monitored using fluorescent labels. The changes in calcium transport were detected using Calcium Green 5-N, the mitochondrial membrane potential and membrane fluidity were elucidated using JC-1 and DPH, respectively. The experiments were carried out during the autumn (October/November) or during the spring (May/June). The obtained data emphasize the effect of seasonal differences on liver mitochondria originating from laboratory animals and outline the importance of planning the experiments during the same seasonal period in order to receive objective and reliable results in the future. Finally, it was revealed the neutral effect of G2.5 dendrimer on mitochondria and its inability to protect mitochondria against overload of calcium ions regardless of seasonality. It was also evidenced that liver mitochondria isolated from autumn-derived animals were more sensitive to calcium and/or dendrimer exposure in comparison with mitochondria isolated from animals investigated during the spring.展开更多
文摘旨在为牛体细胞克隆胚胎体外培养体系的建立奠定基础。利用体细胞核移植技术生产牛重构胚,将重构胚随机分成2组,分别用mSOF和G1.5/G2.5培养,统计2种培养液对牛核移植克隆胚胎发育的影响,用TUNEL荧光检测系统检测2个培养组胚胎细胞凋亡情况,同时用Real time RT-PCR检测热应激蛋白基因Hsp70和凋亡相关基因Bax在2个培养组的相对表达水平。两组的卵裂率分别为85.6%和87.0%、囊胚率分别为22.9%和27.1%,差异都不显著(P>0.05),但G1.5/G2.5组的8-细胞形成率显著高于mSOF组(P<0.05);mSOF组凋亡率为12.1%±1.9%,显著高于G1.5/G2.5组(P<0.05),G1.5/G2.5组的Hsp70和Bax相对表达量显著低于mSOF组(P<0.05)。结果表明,G1.5/G2.5培养液更有利于牛体细胞核移植胚胎的体外发育。
文摘Seasonal variations play an essential role in the metabolism, behavior and activity of the laboratory animals. This study was aimed to examine whether mitochondrial function can be influenced by the seasonal changes and how large is the impact of these fluctuations on experiments with using animal models and further results interpretation. Liver mitochondria were isolated from male Wistar rats and exposed to calcium ions, PAMAM dendrimers G2.5 or their combination: (Ca2+) and dendrimer. The scientific hypothesis assumed that dendrimer G2.5 is able to limit the detrimental effect of Ca2+ ions on mitochondria function, possibly through affecting the following parameters: calcium transport, mitochondrial potential and membrane fluidity. The activity of mitochondria was monitored using fluorescent labels. The changes in calcium transport were detected using Calcium Green 5-N, the mitochondrial membrane potential and membrane fluidity were elucidated using JC-1 and DPH, respectively. The experiments were carried out during the autumn (October/November) or during the spring (May/June). The obtained data emphasize the effect of seasonal differences on liver mitochondria originating from laboratory animals and outline the importance of planning the experiments during the same seasonal period in order to receive objective and reliable results in the future. Finally, it was revealed the neutral effect of G2.5 dendrimer on mitochondria and its inability to protect mitochondria against overload of calcium ions regardless of seasonality. It was also evidenced that liver mitochondria isolated from autumn-derived animals were more sensitive to calcium and/or dendrimer exposure in comparison with mitochondria isolated from animals investigated during the spring.
