首页> 外文期刊>Brain research >Investigation of changes in global gene expression in the frontal cortex of early-weaned and socially isolated piglets using microarray and quantitative real-time RT-PCR.
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Investigation of changes in global gene expression in the frontal cortex of early-weaned and socially isolated piglets using microarray and quantitative real-time RT-PCR.

机译:使用微阵列和定量实时RT-PCR研究早期断奶和社会隔离的仔猪额叶皮质中全球基因表达的变化。

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We hypothesize that early-weaned piglets experience aberrant expression of stress-responsive genes in the frontal cortex, a key brain area involved in cognitive function and behavior organization. To test this hypothesis, female early-weaned piglets (EW; n = 6) were weaned 10 days after birth, while non-weaned piglets (NW; n = 6) were left with their dams. Half of EW (n = 3) and NW (n = 3) animals were socially isolated (SI) for 15 min at 12 days of age, when all animals (n = 12) were euthanized and tissue collected. The effects of EW and SI were examined by gene expression profiling using cDNA microarray hybridizations, generated from a porcine brain cDNA library. A total of 103 genes were differentially expressed (P < 0.05, fold change >1.25) among four direct comparisons. Forty-two genes had known functions, from which 24 showed relevant brain-related functions. Quantitative real-time polymerase chain reaction (Q-RT-PCR) was used to confirm regulation of expression of a subset of 6 genes with important brain functions, selected from the microarray outcomes. In non-weaned animals, a significant suppression of mRNA abundance for carboxypeptidase E, 14-3-3 protein and phosphoprotein enriched in astrocytes 15 kDa was observed in response to SI. Also, in early-weaned animals, diazepam binding inhibitor and actin-related protein 2/3 complex mRNA levels were suppressed in response to SI. Results suggest that social isolation of non- and early-weaned piglets may impact expression of genes involved in regulation of neuronal function, development, and protection in the frontal cortex of young pigs.
机译:我们假设断奶的仔猪在额叶皮层经历了应激反应基因的异常表达,额叶皮层是参与认知功能和行为组织的关键大脑区域。为了验证这一假设,雌性早期断奶仔猪(EW; n = 6)在出生后10天断奶,而未断奶的仔猪(NW; n = 6)则留有母坝。当对所有动物(n = 12)实施安乐死并收集组织时,将一半的EW(n = 3)和NW(n = 3)动物在社会上隔离(SI)15分钟。通过使用从猪脑cDNA文库生成的cDNA微阵列杂交技术,通过基因表达谱分析来检查EW和SI的作用。在四个直接比较中,共有103个基因差异表达(P <0.05,倍数变化> 1.25)。有42个已知功能的基因,其中24个具有相关的大脑相关功能。实时定量聚合酶链反应(Q-RT-PCR)用于确认调控具有重要脑功能的6个基因的子集的表达,这些基因选自微阵列结果。在未断奶的动物中,观察到对SI的响应,羧肽酶E,14-3-3蛋白和富含星形胶质细胞15 kDa的磷蛋白的mRNA丰度得到了显着抑制。同样,在早期断奶的动物中,地西epa结合抑制剂和肌动蛋白相关蛋白2/3复合物mRNA水平受到SI的抑制。结果表明,无断奶仔猪和早期断奶仔猪的社会隔离可能影响幼猪额叶皮质神经元功能,发育和保护相关基因的表达。

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