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Response of Turkey Muscle Satellite Cells to Thermal Challenge. II. Transcriptome Effects in Differentiating Cells

机译:土耳其肌肉卫星细胞对热挑战的反应。二。转录组在分化细胞中的作用

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>Background: Exposure of poultry to extreme temperatures during the critical period of post-hatch growth can seriously affect muscle development and thus compromise subsequent meat quality. This study was designed to characterize transcriptional changes induced in turkey muscle satellite cells by thermal challenge during differentiation. Our goal is to better define how thermal stress alters breast muscle ultrastructure and subsequent development.>Results: Skeletal muscle satellite cells previously isolated from the Pectoralis major muscle of 7-wk-old male turkeys (Meleagris gallopavo) from two breeding lines: the F-line (16 wk body weight-selected) and RBC2 (randombred control line) were used in this study. Cultured cells were induced to differentiate at 38°C (control) or thermal challenge temperatures of 33 or 43°C. After 48 h of differentiation, cells were harvested and total RNA was isolated for RNAseq analysis. Analysis of 39.9 Gb of sequence found 89% mapped to the turkey genome (UMD5.0, annotation 101) with average expression of 18,917 genes per library. In the cultured satellite cells, slow/cardiac muscle isoforms are generally present in greater abundance than fast skeletal isoforms. Statistically significant differences in gene expression were observed among treatments and between turkey lines, with a greater number of genes affected in the F-line cells following cold treatment whereas more differentially expressed (DE) genes were observed in the RBC2 cells following heat treatment. Many of the most significant pathways involved signaling, consistent with ongoing cellular differentiation. Regulation of Ca2+ homeostasis appears to be significantly affected by temperature treatment, particularly cold treatment.>Conclusions: Satellite cell differentiation is directly influenced by temperature at the level of gene transcription with greater effects attributed to selection for fast growth. At lower temperature, muscle-associated genes in the satellite cells were among the genes with the greatest down regulation consistent with slower differentiation and smaller myotubes. Fewer expression differences were observed in the differentiating cells than previously observed for proliferating cells. This suggests the impact of temperature on satellite cells occurs primarily at early points in satellite cell activation.
机译:>背景:在孵化后生长的关键时期,家禽暴露于极端温度会严重影响肌肉发育,从而损害随后的肉品质。这项研究旨在表征在分化过程中热挑战在火鸡肌肉卫星细胞中诱导的转录变化。我们的目标是更好地定义热应激如何改变乳房肌肉的超微结构和随后的发育。>结果:先前从7周龄雄性火鸡(Meleagris gallopavo)的胸大肌中分离出的骨骼肌卫星细胞两条育种系:F系(选择体重16周)和RBC2(随机对照系)。诱导培养的细胞在38°C(对照)或33或43°C的热激发温度下分化。分化48小时后,收获细胞并分离总RNA进行RNAseq分析。分析了39.9 Gb的序列,发现89%定位于火鸡基因组(UMD5.0,注释101),每个文库平均表达18,917个基因。在培养的卫星细胞中,慢/心肌同工型通常比快速骨骼同工型丰度更高。在处理之间和火鸡系之间观察到基因表达的统计学显着差异,冷处理后F系细胞中受影响的基因数量更多,而热处理后RBC2细胞中观察到更多差异表达(DE)基因。许多最重要的途径涉及信号传导,与正在进行的细胞分化一致。 Ca 2 + 动态平衡的调节似乎受温度处理特别是冷处理的影响。>结论:卫星细胞的分化直接受温度在基因转录水平的影响。选择快速增长带来更大的影响。在较低温度下,卫星细胞中与肌肉相关的基因属于下调最大的基因,与分化较慢和肌管较小一致。在分化细胞中观察到的表达差异少于先前在增殖细胞中观察到的差异。这表明温度对卫星细胞的影响主要发生在卫星细胞激活的早期。

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