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单冠白来航鸡中鸡快速骨骼肌肌球蛋白重链亚型的胚胎期转录时间

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

作者信息

Griffin J, St-Pierre N, Lilburn M S, Wick M

机构信息

Department of Animal Sciences, The Ohio State University, Columbus, OH 43201.

Department of Animal Sciences, The Ohio State University, Columbus, OH 43201

出版信息

Poult Sci. 2016 May;95(5):1151-5. doi: 10.3382/ps/pew011. Epub 2016 Feb 16.

Abstract

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.

摘要

有许多因素会显著影响家禽的胚胎发育,因此单纯的孵化天数(按时间顺序计算的年龄)作为研究胚胎生理学的指标并不完美。发育快速的骨骼肌肌球蛋白(MyHC)是胸大肌(PM)中的主要蛋白质,它会按时间顺序表达为一系列高度特异性的发育异构体。在本文所述的研究中,一种新型分子技术(NanoString)被用于表征单冠白来航鸡(SCWL)胚胎胸大肌中肌球蛋白异构体的转录模式。NanoString技术基于通过对目标mRNA转录本进行数字检测和定量来对转录组进行定量分析。从孵化第6天到第19天每天收集胚胎肌肉样本,分离总RNA并使用NanoString对基因转录进行定量。数据在95%置信水平下使用局部加权回归平滑函数进行分析。本研究中获得的MyHC异构体的时间转录情况在更高的特异性和分辨率上与文献一致,从而验证了NanoString可用于基因转录分析。结果支持这样一个假设,即胚胎MyHC异构体的转录模式可用作分子时钟,以进一步研究胚胎快速骨骼肌生长发育背后的发育关系。

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