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Temporal embryonic transcription of chicken fast skeletal myosin heavy chain isoforms in the single comb white leghorn

机译:单梳白来亨鸡中鸡快速骨骼肌球蛋白重链同工型的时间胚胎转录

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摘要

There are numerous factors that can significantly influence embryonic development in poultry and thus make simple days of incubation (chronological age) a less than perfect metric for studying embryonic physiology. The developmental fast skeletal muscle myosin (MyHC), the predominant protein in the Pectoralis major (PM), is temporally expressed as a cadre of highly specific developmental isoforms. In the study described herein, a novel molecular technology (NanoString) was used to characterize the myosin isoform transcriptional patterns in the PM of Single Comb White Leghorn (SCWL) embryos. NanoString technology is based on quantitative analysis of the transcriptome through digital detection and quantification of target mRNA transcripts. Total RNA was isolated and gene transcription quantified using NanoString in embryonic muscle samples collected daily from 6 through 19 days of incubation. Data were analyzed using the LOESS smoothing function at a 95% confidence level. The temporal transcription of MyHC isoforms obtained in this study was consistent with the literature at higher specificity and resolution, thus validating NanoString for use in gene transcription analyses. The results support a hypothesis that the transcription patterns of the embryonic MyHC isoforms may be used as molecular clocks to further investigate the developmental relationships underlying embryonic fast skeletal muscle growth and development.
机译:有很多因素可以显着影响家禽的胚胎发育,因此使简单的孵化天数(按年代划分)不是研究胚胎生理学的理想指标。发育快骨骼肌肌球蛋白(MyHC)是胸大肌(PM)中的主要蛋白质,在时间上被表达为高度特异性发育同工型的干部。在本文所述的研究中,使用了一种新型分子技术(NanoString)来表征单梳白来亨鸡(SCWL)胚胎的PM中的肌球蛋白同种型转录模式。 NanoString技术基于通过对目标mRNA转录物的数字检测和定量对转录组进行定量分析的技术。分离总RNA,并使用NanoString在每天培养6至19天收集的胚胎肌肉样品中定量基因转录。使用LOESS平滑功能以95%的置信度分析数据。在这项研究中获得的MyHC同工型的暂时转录与文献在更高的特异性和分辨率上是一致的,从而验证了NanoString可用于基因转录分析。结果支持一个假说,即胚胎MyHC同工型的转录模式可以用作分子钟,以进一步研究胚胎快速骨骼肌生长与发育之间的发育关系。

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