The present study found that mRNA transcription and protein expression of four adult MyHC
genes all presented some obvious changes with postnatal aging of pigs, especially in the early stage.
The ST muscle, a strongly partitioned muscle, was used for this study because it has the phenotypic expression of the I, IIa, IIb and IIx MyHC
isoforms (Lefaucheur, 2006).
It is well known that maternal nutrient restriction during early to mid-pregnancy had increased the ratio of MyHC
IIb to other isoforms in the longissimus dorsi muscle in the offspring by 17.6% as compared with offspring of ad libitum fed ewes (Zhu et al., 2006).
Therefore, a fast-toslow transition of MyHCs
in SM muscle might be induced by ACL resection, but not RF muscle.
24 h of METF stimuli significantly increased MyHC
protein levels (Figure 4(e)).
The assemblage of the myosin heavy chain protein (MyHC
: isotig02568 and isotig1394) obtained in A.
PCR conditions were as follows: denaturation for each cycle 95[degrees]C (35 cycles, 10 sec for each cycle); annealing both for Myhc
IIa and Atrogin1 60[degrees]C (35 cycles, 0-10 sec for each cycle); for MuRF1 58[degrees]C (35 cycles, 0-10 sec for each cycle); for 18S rRNA 56[degrees]C (35 cycles, 0-10 sec for each cycle); and elongation 72[degrees]C (35 cycles, 4-5 sec for each cycle).
Relative mRNA expressions of myosin heavy-chain (MyHC
) isoform genes in the muscles of Jinjiang (JJ) and F1 SimmentalxJinjiang (SJ) yellow cattle.
For example, increased neuromuscular activity results in a shift in MyHC
isoform from fast to slow muscle fiber (Pette and Staron, 1997), while inactivity leads to a general shift in MyHC
expression and associated metabolic properties along the following line of progression- from type I [right arrow] IIA [right arrow] IIX [right arrow] IIB (Talmadge., 2000).
We evaluated the expression of MyoD, myogenin, and embryonic MyHC
genes by using molecular biological techniques.
According to the isoforms for myosin heavy chain (MyHC
) structure, metabolic patterns and contraction speed, muscle fibers are classified into four categories: slow oxidation type (type I), fast oxidation type (type IIA), fast glycolysis type (type IIB) and intermediate type (type IIC) .
Fixed samples were pretreated with Blocking One (Nacalai Tesque, Kyoto, Japan) for 30 min at RT and incubated with anti-MYOGENIN (diluted 1: 100, Santa Cruz Biotechnology, California, USA), anti-MYOSIN HEAVY CHAIN (MYHC
, diluted 1 : 200, Santa Cruz Biotechnology, California, USA), anti-TRA-1-81 (diluted 1 : 200, Cell Signaling Technology, Massachusetts, USA), anti-SSEA4 (diluted 1: 200, Cell Signaling Technology, Massachusetts, USA), anti-OCT4A (diluted 1 : 200, Cell Signaling Technology, Massachusetts, USA), and anti-NANOG (diluted 1 : 200, Cell Signaling Technology, Massachusetts, USA) antibodies in 5% of Blocking One in PBS with 0.1% Tween20 (PBST) overnight at 4[degrees]C.