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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

Identification and expression of pmMS and ppMS transgenes in mice. (A) Identification of transgenic mice by PCR; (B) Identification of transgenic mice by RT-PCR; (C) Identification of transgenic mice by Southern blot; (D) The mRNA expression level of exogenous pmMS and endogenous myostatin and (E) The mRNA expression level of exogenous ppMS and endogenous myostatin. There were five transgenic mice and six WT (wild type) mice in each group. M, marker; ppMS, wild-type porcine myostatin propeptide; pmMS, porcine myostatin missense mutant; m1, WT male mice; m2, transgenic male mice expressing ppMS; f1, transgenic female mice expressing ppMS; f2, WT female mice; m3, transgenic male mice expressing pmMS; m4, WT male mice; f3, transgenic female mice expressing pmMS; f4, WT female mice. **p < 0.01.
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ijms-16-20020-f001: Identification and expression of pmMS and ppMS transgenes in mice. (A) Identification of transgenic mice by PCR; (B) Identification of transgenic mice by RT-PCR; (C) Identification of transgenic mice by Southern blot; (D) The mRNA expression level of exogenous pmMS and endogenous myostatin and (E) The mRNA expression level of exogenous ppMS and endogenous myostatin. There were five transgenic mice and six WT (wild type) mice in each group. M, marker; ppMS, wild-type porcine myostatin propeptide; pmMS, porcine myostatin missense mutant; m1, WT male mice; m2, transgenic male mice expressing ppMS; f1, transgenic female mice expressing ppMS; f2, WT female mice; m3, transgenic male mice expressing pmMS; m4, WT male mice; f3, transgenic female mice expressing pmMS; f4, WT female mice. **p < 0.01.

Mentions: Both types of transgenic mice were confirmed by PCR and Southern blot (Figure 1A,C). Since there is a difference in gene sequence between C57 BL/6 mice and porcine myostatin, we used reverse transcription PCR by designing primers that only amplify exogenous porcine myostatin to identify the genotype of transgenic mice at the RNA level (Figure 1B). Based on the real-time quantitative PCR (RT-PCR) results, the mRNA expression level of total myostatin, which contains both exogenous porcine and murine endogenous myostatin, was much higher in transgenic mice than in wild-type (WT) mice (Figure 1D,E). Our data confirm that both types of transgenic mice were successfully generated.


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)

Identification and expression of pmMS and ppMS transgenes in mice. (A) Identification of transgenic mice by PCR; (B) Identification of transgenic mice by RT-PCR; (C) Identification of transgenic mice by Southern blot; (D) The mRNA expression level of exogenous pmMS and endogenous myostatin and (E) The mRNA expression level of exogenous ppMS and endogenous myostatin. There were five transgenic mice and six WT (wild type) mice in each group. M, marker; ppMS, wild-type porcine myostatin propeptide; pmMS, porcine myostatin missense mutant; m1, WT male mice; m2, transgenic male mice expressing ppMS; f1, transgenic female mice expressing ppMS; f2, WT female mice; m3, transgenic male mice expressing pmMS; m4, WT male mice; f3, transgenic female mice expressing pmMS; f4, WT female mice. **p < 0.01.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4581338&req=5

ijms-16-20020-f001: Identification and expression of pmMS and ppMS transgenes in mice. (A) Identification of transgenic mice by PCR; (B) Identification of transgenic mice by RT-PCR; (C) Identification of transgenic mice by Southern blot; (D) The mRNA expression level of exogenous pmMS and endogenous myostatin and (E) The mRNA expression level of exogenous ppMS and endogenous myostatin. There were five transgenic mice and six WT (wild type) mice in each group. M, marker; ppMS, wild-type porcine myostatin propeptide; pmMS, porcine myostatin missense mutant; m1, WT male mice; m2, transgenic male mice expressing ppMS; f1, transgenic female mice expressing ppMS; f2, WT female mice; m3, transgenic male mice expressing pmMS; m4, WT male mice; f3, transgenic female mice expressing pmMS; f4, WT female mice. **p < 0.01.
Mentions: Both types of transgenic mice were confirmed by PCR and Southern blot (Figure 1A,C). Since there is a difference in gene sequence between C57 BL/6 mice and porcine myostatin, we used reverse transcription PCR by designing primers that only amplify exogenous porcine myostatin to identify the genotype of transgenic mice at the RNA level (Figure 1B). Based on the real-time quantitative PCR (RT-PCR) results, the mRNA expression level of total myostatin, which contains both exogenous porcine and murine endogenous myostatin, was much higher in transgenic mice than in wild-type (WT) mice (Figure 1D,E). Our data confirm that both types of transgenic mice were successfully generated.

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