Park S K, Gunawan A M, Scheffler T L, Grant A L, Gerrard D E
Department of Animal Sciences, Purdue University, West Lafayette, IN 47907, USA.
J Anim Sci. 2009 Feb;87(2):522-31. doi: 10.2527/jas.2008-1269. Epub 2008 Sep 26.
Genetic selection for improved growth and overall meatiness has resulted in the occurrence of 2 major mutations in pigs, the Rendement Napole (RN) and Halothane (Hal) gene mutations. At the tissue level, these mutations influence energy metabolism in skeletal muscle and muscle fiber type composition, yet also influence total body composition. The RN mutation affects the adenosine monophosphate-activated protein kinase gamma subunit and results in increased glycogen deposition in the muscle, whereas the Hal mutation alters sarcoplasmic calcium release mechanisms and results in altered energy metabolism. From a meat quality standpoint, these mutations independently influence the extent and rate of muscle energy metabolism postmortem, respectively. Even though these mutations alter overall muscle energy metabolism and histochemically derived muscle fiber type independently, their effects have not been yet fully elucidated in respect to myosin heavy chain (MyHC) isoform content and those enzymes responsible for defining energetics of the tissue. Therefore, the objective of this study was to determine the collective effects of the RN and Hal genes on genes and gene products associated with different muscle fiber types in pig skeletal muscle. To overcome potential pitfalls associated with traditional muscle fiber typing, real-time PCR, gel electrophoresis, and Western blotting were used to evaluate MyHC composition and several energy-related gene expressions in muscles from wild-type, RN, Hal, and Hal-RN mutant pigs. The MyHC mRNA levels displayed sequential transitions from IIb to IIx and IIa in pigs bearing the RN mutation. In addition, our results showed MyHC protein isoform abundance is correlated with mRNA level supporting the hypothesis that MyHC genes are transcriptionally controlled. However, transcript abundance of genes involved in energy metabolism, including lactate dehydrogenase, citrate synthase, glycogen synthase, and peroxisome proliferator-activated receptor alpha, was not different between genotypes. These data show that the RN and Hal gene mutations alter muscle fiber type composition and suggest that muscle fiber energy metabolism and speed of contraction, the 2 determinants of muscle fiber type, can be uncoupled.
为提高生长速度和整体肉质进行的基因选择,导致猪出现了两个主要突变,即拿破仑效率(RN)和氟烷(Hal)基因突变。在组织水平上,这些突变影响骨骼肌中的能量代谢和肌纤维类型组成,但也会影响全身组成。RN突变影响腺苷单磷酸激活的蛋白激酶γ亚基,导致肌肉中糖原沉积增加,而Hal突变改变肌浆钙释放机制,导致能量代谢改变。从肉质角度来看,这些突变分别独立影响宰后肌肉能量代谢的程度和速率。尽管这些突变分别独立改变整体肌肉能量代谢和组织化学衍生的肌纤维类型,但它们对肌球蛋白重链(MyHC)异构体含量以及负责定义组织能量学的那些酶的影响尚未完全阐明。因此,本研究的目的是确定RN和Hal基因对猪骨骼肌中与不同肌纤维类型相关的基因和基因产物的综合影响。为了克服与传统肌纤维分型相关的潜在缺陷,使用实时PCR、凝胶电泳和蛋白质免疫印迹法来评估野生型、RN、Hal和Hal-RN突变猪肌肉中的MyHC组成和几种能量相关基因的表达。携带RN突变的猪中,MyHC mRNA水平呈现从IIb到IIx再到IIa的顺序转变。此外,我们的结果表明MyHC蛋白异构体丰度与mRNA水平相关,支持MyHC基因受转录调控的假设。然而,参与能量代谢的基因,包括乳酸脱氢酶、柠檬酸合酶、糖原合酶和过氧化物酶体增殖物激活受体α的转录本丰度在不同基因型之间没有差异。这些数据表明,RN和Hal基因突变改变了肌纤维类型组成,并表明肌纤维能量代谢和收缩速度这两个肌纤维类型的决定因素可能是解偶联的。