Limits...
Over-Expression of Porcine Myostatin Missense Mutant Leads to A Gender Difference in Skeletal Muscle Growth between Transgenic Male and Female Mice.

Ma D, Gao P, Qian L, Wang Q, Cai C, Jiang S, Xiao G, Cui W - Int J Mol Sci (2015)

Bottom Line: Piedmontese cattle breeds have a missense mutation, which results in a cysteine to tyrosine substitution in the mature myostatin protein (C313Y).This loss-of-function mutation in myostatin results in a double-muscled phenotype in cattle.These results provide useful insight and basic theory to future studies on improving pork quality by genetically manipulating myostatin expression or by regulating myostatin activity.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory for Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China. madezun@126.com.

ABSTRACT
Myostatin, a transforming growth factor-β family member, is a negative regulator of skeletal muscle development and growth. Piedmontese cattle breeds have a missense mutation, which results in a cysteine to tyrosine substitution in the mature myostatin protein (C313Y). This loss-of-function mutation in myostatin results in a double-muscled phenotype in cattle. Myostatin propeptide is an inhibitor of myostatin activity and is considered a potential agent to stimulate muscle growth in livestock. In this study, we generated transgenic mice overexpressing porcine myostatin missense mutant (pmMS), C313Y, and wild-type porcine myostatin propeptide (ppMS), respectively, to examine their effects on muscle growth in mice. Enhanced muscle growth was observed in both pmMS and ppMS transgenic female mice and also in ppMS transgenic male mice. However, there was no enhanced muscle growth observed in pmMS transgenic male mice. To explore why there is such a big difference in muscle growth between pmMS and ppMS transgenic male mice, the expression level of androgen receptor (AR) mutant AR45 was measured by Western blot. Results indicated that AR45 expression significantly increased in pmMS transgenic male mice while it decreased dramatically in ppMS transgenic male mice. Our data demonstrate that both pmMS and ppMS act as myostatin inhibitors in the regulation of muscle growth, but the effect of pmMS in male mice is reversed by an increased AR45 expression. These results provide useful insight and basic theory to future studies on improving pork quality by genetically manipulating myostatin expression or by regulating myostatin activity.

No MeSH data available.


Related in: MedlinePlus

Protein expression level of AR45 and myostatin in the gastrocnemius of transgenic male mice and WT male mice by Western blot. (A) Protein expression level of AR45 (67 kDa) in pmMS transgenic male and WT male mice; (B) Protein expression level of AR45 (67 kDa) in ppMS transgenic male and WT male mice; (C) Protein expression level of myostatin (26 kDa) in pmMS transgenic male and WT male mice; (D) Protein expression level of myostatin (26 kDa) in ppMS transgenic male and WT male mice. In panels (A–D), each lane contains a sample from one individual mouse; (E) Quantitation of AR45 protein band intensity by densitometry using image J software and (F) Quantitation of myostatin protein band intensity by densitometry using image J software (NIH, Bethesda, MD, USA). GAPDH was used as an internal reference. MSTN, myostatin; *p < 0.05, **p < 0.01.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4581338&req=5

ijms-16-20020-f004: Protein expression level of AR45 and myostatin in the gastrocnemius of transgenic male mice and WT male mice by Western blot. (A) Protein expression level of AR45 (67 kDa) in pmMS transgenic male and WT male mice; (B) Protein expression level of AR45 (67 kDa) in ppMS transgenic male and WT male mice; (C) Protein expression level of myostatin (26 kDa) in pmMS transgenic male and WT male mice; (D) Protein expression level of myostatin (26 kDa) in ppMS transgenic male and WT male mice. In panels (A–D), each lane contains a sample from one individual mouse; (E) Quantitation of AR45 protein band intensity by densitometry using image J software and (F) Quantitation of myostatin protein band intensity by densitometry using image J software (NIH, Bethesda, MD, USA). GAPDH was used as an internal reference. MSTN, myostatin; *p < 0.05, **p < 0.01.

Mentions: To explore why there is such a huge difference in muscle growth and fiber size between pmMS and ppMS transgenic male mice, the AR45 protein level was analyzed by Western blot. The intensity of the AR45 protein band in each genotype of male mice was measured by densitometry using Image J 1.48u (NIH, Bethesda, MD, USA). Results indicated that compared with WT male mice, the level of AR45 protein increased 32.65% in pmMS transgenic male mice but decreased 46.07% in ppMS transgenic male mice (Figure 4A,B,E). On the other hand, the AR45 level is only slightly higher in pmMS female mice than in wild-type female mice (Figure 5A); the difference in the AR45 level between pmMS female mice and wild-type female mice is not significant.


Over-Expression of Porcine Myostatin Missense Mutant Leads to A Gender Difference in Skeletal Muscle Growth between Transgenic Male and Female Mice.

Ma D, Gao P, Qian L, Wang Q, Cai C, Jiang S, Xiao G, Cui W - Int J Mol Sci (2015)

Protein expression level of AR45 and myostatin in the gastrocnemius of transgenic male mice and WT male mice by Western blot. (A) Protein expression level of AR45 (67 kDa) in pmMS transgenic male and WT male mice; (B) Protein expression level of AR45 (67 kDa) in ppMS transgenic male and WT male mice; (C) Protein expression level of myostatin (26 kDa) in pmMS transgenic male and WT male mice; (D) Protein expression level of myostatin (26 kDa) in ppMS transgenic male and WT male mice. In panels (A–D), each lane contains a sample from one individual mouse; (E) Quantitation of AR45 protein band intensity by densitometry using image J software and (F) Quantitation of myostatin protein band intensity by densitometry using image J software (NIH, Bethesda, MD, USA). GAPDH was used as an internal reference. MSTN, myostatin; *p < 0.05, **p < 0.01.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581338&req=5

ijms-16-20020-f004: Protein expression level of AR45 and myostatin in the gastrocnemius of transgenic male mice and WT male mice by Western blot. (A) Protein expression level of AR45 (67 kDa) in pmMS transgenic male and WT male mice; (B) Protein expression level of AR45 (67 kDa) in ppMS transgenic male and WT male mice; (C) Protein expression level of myostatin (26 kDa) in pmMS transgenic male and WT male mice; (D) Protein expression level of myostatin (26 kDa) in ppMS transgenic male and WT male mice. In panels (A–D), each lane contains a sample from one individual mouse; (E) Quantitation of AR45 protein band intensity by densitometry using image J software and (F) Quantitation of myostatin protein band intensity by densitometry using image J software (NIH, Bethesda, MD, USA). GAPDH was used as an internal reference. MSTN, myostatin; *p < 0.05, **p < 0.01.
Mentions: To explore why there is such a huge difference in muscle growth and fiber size between pmMS and ppMS transgenic male mice, the AR45 protein level was analyzed by Western blot. The intensity of the AR45 protein band in each genotype of male mice was measured by densitometry using Image J 1.48u (NIH, Bethesda, MD, USA). Results indicated that compared with WT male mice, the level of AR45 protein increased 32.65% in pmMS transgenic male mice but decreased 46.07% in ppMS transgenic male mice (Figure 4A,B,E). On the other hand, the AR45 level is only slightly higher in pmMS female mice than in wild-type female mice (Figure 5A); the difference in the AR45 level between pmMS female mice and wild-type female mice is not significant.

Bottom Line: Piedmontese cattle breeds have a missense mutation, which results in a cysteine to tyrosine substitution in the mature myostatin protein (C313Y).This loss-of-function mutation in myostatin results in a double-muscled phenotype in cattle.These results provide useful insight and basic theory to future studies on improving pork quality by genetically manipulating myostatin expression or by regulating myostatin activity.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory for Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China. madezun@126.com.

ABSTRACT
Myostatin, a transforming growth factor-β family member, is a negative regulator of skeletal muscle development and growth. Piedmontese cattle breeds have a missense mutation, which results in a cysteine to tyrosine substitution in the mature myostatin protein (C313Y). This loss-of-function mutation in myostatin results in a double-muscled phenotype in cattle. Myostatin propeptide is an inhibitor of myostatin activity and is considered a potential agent to stimulate muscle growth in livestock. In this study, we generated transgenic mice overexpressing porcine myostatin missense mutant (pmMS), C313Y, and wild-type porcine myostatin propeptide (ppMS), respectively, to examine their effects on muscle growth in mice. Enhanced muscle growth was observed in both pmMS and ppMS transgenic female mice and also in ppMS transgenic male mice. However, there was no enhanced muscle growth observed in pmMS transgenic male mice. To explore why there is such a big difference in muscle growth between pmMS and ppMS transgenic male mice, the expression level of androgen receptor (AR) mutant AR45 was measured by Western blot. Results indicated that AR45 expression significantly increased in pmMS transgenic male mice while it decreased dramatically in ppMS transgenic male mice. Our data demonstrate that both pmMS and ppMS act as myostatin inhibitors in the regulation of muscle growth, but the effect of pmMS in male mice is reversed by an increased AR45 expression. These results provide useful insight and basic theory to future studies on improving pork quality by genetically manipulating myostatin expression or by regulating myostatin activity.

No MeSH data available.


Related in: MedlinePlus