Department of Food Science and Human Nutrition, Michigan State University, East Lansing, Michigan 48824, USA.
BMC Genomics. 2011 Mar 8;12:143. doi: 10.1186/1471-2164-12-143.
Skeletal muscle growth and development from embryo to adult consists of a series of carefully regulated changes in gene expression. Understanding these developmental changes in agriculturally important species is essential to the production of high quality meat products. For example, consumer demand for lean, inexpensive meat products has driven the turkey industry to unprecedented production through intensive genetic selection. However, achievements of increased body weight and muscle mass have been countered by an increased incidence of myopathies and meat quality defects. In a previous study, we developed and validated a turkey skeletal muscle-specific microarray as a tool for functional genomics studies. The goals of the current study were to utilize this microarray to elucidate functional pathways of genes responsible for key events in turkey skeletal muscle development and to compare differences in gene expression between two genetic lines of turkeys. To achieve these goals, skeletal muscle samples were collected at three critical stages in muscle development: 18d embryo (hyperplasia), 1d post-hatch (shift from myoblast-mediated growth to satellite cell-modulated growth by hypertrophy), and 16 wk (market age) from two genetic lines: a randombred control line (RBC2) maintained without selection pressure, and a line (F) selected from the RBC2 line for increased 16 wk body weight. Array hybridizations were performed in two experiments: Experiment 1 directly compared the developmental stages within genetic line, while Experiment 2 directly compared the two lines within each developmental stage.
A total of 3474 genes were differentially expressed (false discovery rate; FDR < 0.001) by overall effect of development, while 16 genes were differentially expressed (FDR < 0.10) by overall effect of genetic line. Ingenuity Pathways Analysis was used to group annotated genes into networks, functions, and canonical pathways. The expression of 28 genes involved in extracellular matrix regulation, cell death/apoptosis, and calcium signaling/muscle function, as well as genes with miscellaneous function was confirmed by qPCR.
The current study identified gene pathways and uncovered novel genes important in turkey muscle growth and development. Future experiments will focus further on several of these candidate genes and the expression and mechanism of action of their protein products.
从胚胎到成年,骨骼肌的生长和发育包括一系列精心调控的基因表达变化。了解农业上重要物种的这些发育变化对于生产高质量的肉类产品至关重要。例如,消费者对瘦肉、廉价肉产品的需求促使火鸡产业通过密集的遗传选择达到了前所未有的产量。然而,体重和肌肉质量的增加却伴随着肌病和肉质缺陷发生率的增加。在之前的一项研究中,我们开发并验证了一种火鸡骨骼肌特异性微阵列,作为功能基因组学研究的工具。本研究的目的是利用该微阵列阐明负责火鸡骨骼肌发育关键事件的基因的功能途径,并比较两种火鸡遗传系之间的基因表达差异。为了实现这些目标,在肌肉发育的三个关键阶段采集骨骼肌样本:18d 胚胎(增生)、1d 出壳(由成肌细胞介导的生长向卫星细胞调节的生长转变)和 16 周(市场年龄)来自两个遗传系:未经选择压力维持的随机杂交系(RBC2)和从 RBC2 系中选择的增加 16 周体重的系(F)。在两个实验中进行了微阵列杂交:实验 1 直接比较遗传系内的发育阶段,而实验 2 直接比较每个发育阶段内的两个系。
共有 3474 个基因因发育的总体效应而差异表达(错误发现率; FDR < 0.001),而 16 个基因因遗传系的总体效应而差异表达(FDR < 0.10)。通路分析工具(Ingenuity Pathways Analysis)用于将注释基因分组为网络、功能和经典途径。通过 qPCR 验证了 28 个参与细胞外基质调节、细胞死亡/凋亡和钙信号转导/肌肉功能的基因以及具有多种功能的基因的表达。
本研究确定了与火鸡肌肉生长和发育相关的基因途径,并发现了一些新的重要基因。未来的实验将进一步集中在这些候选基因中的几个基因及其蛋白质产物的表达和作用机制上。