Myostatin short interfering hairpin RNA gene transfer increases skeletal muscle mass
| Autor: | Jacob Rajfer, Suzanne Reisz-Porszasz, Monica G. Ferrini, Freddi I. Zuniga, Thomas R. Magee, Dolores Vernet, Liliana Cantini, Nestor F. Gonzalez-Cadavid, Jorge N. Artaza |
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| Rok vydání: | 2006 |
| Předmět: |
Male
Genetic Vectors Gene Expression Myostatin Transfection Cell Line Muscle hypertrophy Small hairpin RNA Mice Genes Reporter Transforming Growth Factor beta Drug Discovery Myosin Gene expression Genetics medicine Animals Humans Gene Silencing RNA Small Interfering Muscle Skeletal Molecular Biology Genetics (clinical) Base Sequence biology Gene Transfer Techniques Skeletal muscle musculoskeletal system Molecular biology Rats Inbred F344 Muscle atrophy Rats Blot Electroporation medicine.anatomical_structure Lac Operon biology.protein Molecular Medicine medicine.symptom Plasmids |
| Zdroj: | The Journal of Gene Medicine. 8:1171-1181 |
| ISSN: | 1521-2254 1099-498X |
| DOI: | 10.1002/jgm.946 |
| Popis: | Background Myostatin negatively regulates skeletal muscle growth. Myostatin knockout mice exhibit muscle hypertrophy and decreased interstitial fibrosis. We investigated whether a plasmid expressing a short hairpin interfering RNA (shRNA) against myostatin and transduced using electroporation would increase local skeletal muscle mass. Methods Short interfering RNAs (siRNAs) targeting myostatin were co-transfected with a myostatin-expressing plasmid into HEK293 cells and identified for myostatin silencing by Western blot. Corresponding shRNAs were cloned into plasmid shRNA expression vectors. Myostatin or a randomer negative control shRNA plasmid was injected and electroporated into the tibialis anterior or its contralateral muscle, respectively, of nine rats that were sacrificed after 2 weeks. Six other rats received a β-galactosidase reporter plasmid and were sacrificed at 1, 2, and 4 weeks. Uptake of plasmid was examined by β-galactosidase expression, whereas myostatin expression was determined by real-time polymerase chain reaction (PCR) and Western blotting. Muscle fiber size was determined by histochemistry. Satellite cell proliferation was determined by PAX7 immunohistochemistry. Myosin heavy chain type II (MHCII) expression was determined by Western blot. Results β-Galactosidase reporter plasmid was expressed at 1 and 2 weeks but diminished by 4 weeks in tibialis anterior skeletal muscle. Myostatin shRNA reduced myostatin mRNA and protein expression by 27 and 48%, respectively. Tibialis anterior weight, fiber size, and MHCII increased by 10, 34, and 38%, respectively. Satellite cell number was increased by over 2-fold. Conclusions This is the first demonstration that myostatin shRNA gene transfer is a potential strategy to increase muscle mass. Copyright © 2006 John Wiley & Sons, Ltd. |
| Databáze: | OpenAIRE |
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