Myostatin inactivation induces a similar muscle molecular signature in double-muscled cattle as in mice (original) (raw)
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Target genes of myostatin loss-of-function in muscles of late bovine fetuses
BMC Genomics, 2007
Background: Myostatin, a muscle-specific member of the Transforming Growth Factor beta family, negatively regulates muscle development. Double-muscled (DM) cattle have a loss-offunction mutation in their myostatin gene responsible for the hypermuscular phenotype. Thus, these animals are a good model for understanding the mechanisms underpinning muscular hypertrophy. In order to identify individual genes or networks that may be myostatin targets, we looked for genes that were differentially expressed between DM and normal (NM) animals (n = 3 per group) in the semitendinosus muscle (hypertrophied in DM animals) at 260 days of fetal development (when the biochemical differentiation of muscle is intensive). A heterologous microarray (human and murine oligonucleotide sequences) of around 6,000 genes expressed in muscle was used.
Proteomic analysis of bovine skeletal muscle hypertrophy
PROTEOMICS, 2005
Myostatin plays a major role in muscle growth and development and animals with disruption of this gene display marked increases in muscle mass. Little is known about muscle physiological adaptations in relation to this muscle hypertrophy. To provide a more comprehensive view, we analyzed bovine muscles from control, heterozygote and homozygote young Belgian blue bulls for myostatin deletion, which results in a normal level of inactive myostatin. Heterozygote and homozygote animals were characterized by a higher proportion of fast-twitch glycolytic fibers in Semitendinosus muscle. Differential proteomic analysis of this muscle was performed using two-dimensional gel electrophoresis followed by mass spectrometry. Thirteen proteins, corresponding to 28 protein spots, were significantly altered in response to the myostatin deletion. The observed changes in protein expression are consistent with an increased fast muscle phenotype, suggesting that myostatin negatively controls mainly fast-twitch glycolytic fiber number. Finally, we demonstrated that differential mRNA splicing of fast troponin T is altered by the loss of myostatin function. The structure of mutually exclusive exon 16 appears predominantly expressed in muscles from heterozygote and homozygote animals. This suggests a role for exon 16 of fast troponin T in the physiological adaptation of the fast muscle phenotype.
Genetics Research, 2004
Myostatin is a negative regulator of muscle growth, and mutations in this gene are associated with muscular hypertrophy and reduced fat in mice, humans and cattle. Marker assisted introgression (MAI) was used to introduce a murine myostatin mutation, Mstn Cmpt-dl1Abc (Compact, C), into a high growth mouse line (DUHi) Objective: To investigate the effects of myostatin-deficiency in a high growth background on growth, body composition and, using samples from M. rectus femoris and M. longissimus dorsi, on muscle histology. Results: Inter se matings between C/+ animals up to generation 10 produced a total of 838 genotyped pups, 29% +/+, 63% +/C, and only 8% C/C. This highly significant distortion of the segregation ratio shows that Compact is associated with lower fitness on the DUHi-background. Homozygous (C/C) mice were, compared to wildtype (+/+) mice, c4-5% lighter, had c7-8% shorter tails, increased muscle weights (e.g. M. quadriceps in males was 59% heavier), an increased 'dressing percentage' (c49 vs. 39%) and the weights of several organs (e.g. liver, kidney, heart) were significantly reduced (12 -20%). Total body fat content was higher in +/+ animals (c17.5%) than in C/C-animals (10-12%). Total muscle fibre number was increased by 24%, whereas fibre size was not significantly affected. Protein and DNA concentrations, DNA/protein ratios and specific CK activity remained unchanged, indicative of increases in the contents of total DNA and muscle specific protein with increased muscle mass. Fibre type distribution was shifted to white glycolytic muscle fibres (+c16% units) at the expense of red oxidative fibres. Capillary density was substantially lower in C/C than in +/+ mice, with lower number of capillaries per fibre (-35%) and larger fibre area per capillary (+77%).
BMC Genomics, 2009
Background Myostatin (MSTN), a member of the TGF-β superfamily, has been identified as a negative regulator of skeletal muscle mass. Inactivating mutations in the MSTN gene are responsible for the development of a hypermuscular phenotype. In this study, we performed transcriptomic and proteomic analyses to detect altered expression/abundance of genes and proteins. These differentially expressed genes and proteins may represent new molecular targets of MSTN and could be involved in the regulation of skeletal muscle mass. Results Transcriptomic analysis of the Quadriceps muscles of 5-week-old MSTN-null mice (n = 4) and their controls (n = 4) was carried out using microarray (human and murine oligonucleotide sequences) of 6,473 genes expressed in muscle. Proteomic profiles were analysed using two-dimensional gel electrophoresis coupled with mass spectrometry. Comparison of the transcriptomic profiles revealed 192 up- and 245 down- regulated genes. Genes involved in the PI3K pathway, insulin/IGF pathway, carbohydrate metabolism and apoptosis regulation were up-regulated. Genes belonging to canonical Wnt, calcium signalling pathways and cytokine-receptor cytokine interaction were down-regulated. Comparison of the protein profiles revealed 20 up- and 18 down-regulated proteins spots. Knockout of the MSTN gene was associated with up-regulation of proteins involved in glycolytic shift of the muscles and down-regulation of proteins involved in oxidative energy metabolism. In addition, an increased abundance of survival/anti-apoptotic factors were observed. Conclusion All together, these results showed a differential expression of genes and proteins related to the muscle energy metabolism and cell survival/anti-apoptotic pathway (e.g. DJ-1, PINK1, 14-3-3ε protein, TCTP/GSK-3β). They revealed the PI3K and apoptotic pathways as MSTN targets and are in favour of a role of MSTN as a modulator of cell survival in vivo.
Australian Journal of Agricultural Research, 2006
This study investigated whether the expression profile of GDF8 (myostatin), myogenic regulatory factors (MRFs: MYF5, MYOD1, MYOG (myogenin), and MYF6), and IGF-system (IGF1, IGF2, IGF1R) genes are correlated with anatomical muscle, nutrition level, and estimated breeding values (EBVs) for muscling, growth, and/or fatness. Real-time PCR was employed to quantitatively measure the mRNA levels of these genes in the semimembranosus (SM) and semitendinosus (ST) muscles of growing lambs. The lambs were sired by Poll Dorset rams with differing EBVs for growth, muscling, and fatness, and were fed either high or low quality and availability pasture from birth to ∼8 months of age. With the exception of MYOD1, the mRNA levels of all genes examined in this study showed varying degrees of nutritional regulation. All the MRF mRNA levels were higher in the SM muscle than the ST muscle, whereas myostatin mRNA was higher in the ST muscle than the SM muscle. Interactions between muscle type and nutrition were detected for IGF2, MYF6, and myogenin, while positive correlations between IGF2 and IGF1R and between MYOD1 and myogenin mRNA levels were apparent in both muscles. At the genotypic level, subtle differences in mRNA levels suggested interactions between nutrition and sire EBV. The findings of this study confirm that the MRFs, IGFs, and myostatin genes are differentially affected by a variety of factors that include nutrition, muscle type, and sire EBVs. Together, these data suggest that this suite of genes has important roles during postnatal muscle growth, even at quite late stages of growth and development.
Animal Genetics, 2008
Myostatin (MSTN), a transforming growth factor b superfamily member, is an essential factor for the growth and development of muscle mass. The protein functions as a negative regulator of muscle growth and is related to the so-called double-muscling phenotype in cattle, where a series of mutations renders the gene inactive. One particular breed of pigs, the Belgian Piétrain, also shows a heavily muscled phenotype. The similarity of muscular phenotypes between the double-muscled cattle and Piétrain pigs indicated that MSTN may be a candidate gene for muscular hypertrophy in pigs. In this study, we sequenced and analysed the complete MSTN gene from 45 pigs of five different breeds, including the heavily muscled Piétrain breed at one extreme and the Meishan and Wild boar breeds at the other extreme. In total, 7626 bp of the porcine MSTN gene were sequenced, including the 5¢ and 3¢ UTR. Fifteen polymorphic loci were found, three of which were located in the promoter region, five in intron 1 and seven in intron 2. Most mutations were found when comparing the obtained MSTN sequence with porcine MSTN sequences already published. However, one polymorphism located at position 447 of the porcine MSTN promoter had a very high allele frequency in the Piétrain pig breed and disrupted a putative myocyte enhancer factor 3 binding site. Real-time PCR using Sybr Green showed that this mutation was associated with expression levels of the MSTN gene in m. longissimus dorsi at an age of 4 weeks.
Proliferation Rates of Bovine Primary Muscle Cells Relate to Liveweight and Carcase Weight in Cattle
PloS one, 2015
Muscling in cattle is largely influenced by genetic background, ultimately affecting beef yield and is of major interest to the beef industry. This investigation aimed to determine whether primary skeletal muscle cells isolated from different breeds of cattle with a varying genetic potential for muscling differ in their myogenic proliferative capacity. Primary skeletal muscle cells were isolated and cultured from the Longissimus muscle (LM) of 6 month old Angus, Hereford and Wagyu X Angus cattle. Cells were assessed for rate of proliferation and gene expression of PAX7, MYOD, MYF5, and MYOG. Proliferation rates were found to differ between breeds of cattle whereby myoblasts from Angus cattle were found to proliferate at a greater rate than those of Hereford and Wagyu X Angus during early stages of growth (5-20 hours in culture) in vitro (P < 0.05). The proliferation rates of myoblasts during early stages of culture in vitro were also found to be positively related to the liveweig...
Domestic Animal Endocrinology, 2014
The purpose of this study was to determine whether myostatin alters glucose transporter-4 (GLUT4) expression in bovine skeletal muscles and myoblasts isolated from doublemuscled (DM) and normal-muscled (NM) Japanese Shorthorn cattle. Plasma concentrations of glucose were lower in DM cattle than in NM cattle (P < 0.01). The expression of GLUT4 messenger RNA (mRNA) in the skeletal muscle ex vivo and in myoblasts at 72 h after differentiation in vitro was higher in DM cattle than in NM cattle (P < 0.01). In contrast, the NM and DM cattle did not differ with respect to skeletal muscle expression of GLUT1 and myocyte enhancer factor-2c (MEF2c), a transcription factor of GLUT4. In differentiated myoblasts, the expression of GLUT1, GLUT4, and MEF2c mRNAs was greater in DM cattle than in NM cattle (P < 0.01). In the presence and absence of insulin, glucose uptake in myoblasts was increased in DM cattle relative to that of NM cattle (P < 0.01). The addition of myostatin decreased the expression of GLUT4 and MEF2c mRNAs in DM myoblasts (P < 0.05). Results of the present study suggest that myostatin inhibits the expression of GLUT4 mRNA possibly via MEF2c and that the greater ability of the DM cattle to produce muscle relative to the NM cattle may be due to their greater sensitivity to insulin and greater use of glucose.
Inferring the in vivo cellular program of developing bovine skeletal muscle from expression data
Gene Expression Patterns, 2013
We outline an in vivo cellular program of bovine longissimus muscle development inferred from expression data from 60 days post conception to 3 months postnatal. Analytic challenges included changes in cellular composition, ambiguous 'diagnostic' markers of cell type and contrasts between cattle human and mouse myogenesis. Nevertheless, the expression profiles of the myosin isoforms support slow and fast muscle fibres emanating from primary and secondary myogenesis respectively, while expression of the prenatal myosin subunits is down regulated prior to birth. Of the canonical pro-myogenic transcription factors (TF), MYF6 and MYF5 are negatively co-expressed, with MYF6 displaying higher expression in the post-natal samples and MYF5, MYOG, HES6 and PAX7 displaying higher expression in early development. A set of TFs (SIX1, EYA2 and DACH2) considered important in undifferentiated murine cells were equally abundant in differentiated bovine cells. An examination of mammalian regulators of fibre composition, muscle mass and muscle metabolism, underscored the roles of PPARGC1A, TGFb signalling and the NHR4 Nuclear Hormone Receptors on bovine muscle development. Enriched among the most variably expressed genes from the entire data set were molecules regulating mitochondrial metabolism of carbohydrate (PDK4), fat (UCP3), protein (AGXT2L1) and high energy phosphate (CKMT2). The dramatic increase in the expression of these transcripts, which may enable the peri-natal transition to metabolic independence critical for new-born herbivores, provides surprising evidence for substantial developmental remodelling of muscle mitochondria and reflects changes in nutrient availability. Overall, despite differences in size, metabolism and physiology, the muscle structural subunit expression program appears very similar in ruminants, rodents and humans.