Ectopic Overexpression of Porcine Myh1 Increased in Slow Muscle Fibers and Enhanced Endurance Exercise in Transgenic Mice
Abstract
:1. Introduction
2. Results
2.1. Identification of Porcine Myh1 Gene
2.2. Establishment of Ectopic Overexpression of Porcine Myh1 in Mice
2.3. Effects on Slow Fiber Muscle in Myh1 TG Mice
2.4. Effect of Ectopic Expressed Porcine Myh1 to Muscle Type Formation
2.5. Enhanced Physical Performance in Myh1 TG Mice
3. Discussion
4. Materials and Methods
4.1. Use of Mice and IACUC Commitment
4.2. Construct of Myh1 Overexpression Vector and TG Mice Production
4.3. Analysis of Gene Expression
4.4. Western Blotting
4.5. Immunohistochemistry
4.6. ATPase and Succinate Dehydrogenase Staining
4.7. Physiological Studies
4.8. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Gene | Primer Sequence (5’-3’) | Size (bp) | |
---|---|---|---|
Forward | Reverse | ||
Myh7 (mouse) | AGTCCCAGGTCAACAAGCTG | TTCCACCTAAAGGGCTGTTG | 145 |
Myh2 (mouse) | AGTCCCAGGTCAACAAGCTG | GCATGACCAAAGGTTTCACA | 130 |
Myh1 (mouse) | AGTCCCAGGTCAACAAGCTG | CACATTTTGCTCATCTCTTTG | 113 |
Myh1 (porcine) | CTTCACTGGCGCAGCAGGT | AGATGCGGATGCCCTCCA | 256 |
Myh4 (mouse) | AGTCCCAGGTCAACAAGCTG | TTTCTCCTGTCACCTCTCAACA | 100 |
Myoglobin (mouse) | GCAAGGCCCTGGAGCTCTTC | GCTTGGTGGGCTGGACAGTG | 100 |
Tnnt1 (mouse) | CCCCCGAAGATTCCAGAAGG | TGCGGTCTTTTAGTGCAATGAG | 154 |
Tnni1 (mouse) | ATGCCGGAAGTTGAGAGGAAA | TCCGAGAGGTAACGCACCTT | 140 |
Tnnc1 (mouse) | GCGGTAGAACAGTTGACAGAG | CCAGCTCCTTGGTGCTGAT | 103 |
Aldoa (mouse) | GGCAACACCCAGCAACAGAC | TCATCTGCAGCCAGGATGCC | 158 |
Pvalb (mouse) | ATCAAGAAGGCGATAGGAGCC | GGCCAGAAGCGTCTTTGTT | 252 |
Tnnt3 (mouse) | GGAACGCCAGAACAGATTGG | TGGAGGACAGAGCCTTTTTCTT | 104 |
Tnni2 (mouse) | AGAGTGTGATGCTCCAGATAGC | AGCAACGTCGATCTTCGCA | 170 |
Tnnc2 (mouse) | ATGGCAGCGGTACTATCGACT | CCTTCGCATCCTCTTTCATCTG | 72 |
GAPDH (mouse) | TGAAGGTCGGTGTGAACGG | CGTGAGTGGAGTCATACTGGAA | 150 |
GAPDH (porcine) | CCTTCATTGACCTCAACTACAT | CCAAAGTTGTCATGGATGACC | 400 |
Antibody | Number | Source |
---|---|---|
Myc-Tag | #2276 | Cell Signaling |
MYH4 | SC-168672 | Santa Cruz Biotechnology |
MYH7 | SC-53089 | Santa Cruz Biotechnology |
Myoglobin | SC-8081 | Santa Cruz Biotechnology |
Troponin I-SS | SC-8119 | Santa Cruz Biotechnology |
Cytochrom C | SC-13156 | Santa Cruz Biotechnology |
β-actin | #4970 | Cell Signaling |
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Ahn, J.S.; Kim, D.-H.; Park, H.-B.; Han, S.-H.; Hwang, S.; Cho, I.-C.; Lee, J.-W. Ectopic Overexpression of Porcine Myh1 Increased in Slow Muscle Fibers and Enhanced Endurance Exercise in Transgenic Mice. Int. J. Mol. Sci. 2018, 19, 2959. https://doi.org/10.3390/ijms19102959
Ahn JS, Kim D-H, Park H-B, Han S-H, Hwang S, Cho I-C, Lee J-W. Ectopic Overexpression of Porcine Myh1 Increased in Slow Muscle Fibers and Enhanced Endurance Exercise in Transgenic Mice. International Journal of Molecular Sciences. 2018; 19(10):2959. https://doi.org/10.3390/ijms19102959
Chicago/Turabian StyleAhn, Jin Seop, Dong-Hwan Kim, Hee-Bok Park, Sang-Hyun Han, Seongsoo Hwang, In-Cheol Cho, and Jeong-Woong Lee. 2018. "Ectopic Overexpression of Porcine Myh1 Increased in Slow Muscle Fibers and Enhanced Endurance Exercise in Transgenic Mice" International Journal of Molecular Sciences 19, no. 10: 2959. https://doi.org/10.3390/ijms19102959