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Analysis of gene expression during the onset of muscle hypertrophy in callipyge lambs: time course

ABSTRACT: The callipyge mutation causes postnatal muscle hypertrophy in heterozygous lambs that inherit a paternal callipyge allele (+/CLPG). Our hypothesis was that the up-regulation of one or both of the affected paternally expressed genes (DLK1 or PEG11) initiates changes in biochemical and physiological pathways in skeletal muscle to induce hypertrophy. The goal of this study was to identify changes in gene expression during the onset of muscle hypertrophy in order to identify the pathways that are involved in the expression of the callipyge phenotype. Gene expression was analyzed in longissimus dorsi total RNA from lambs at 10, 20, and 30 days of age using the Affymetrix Bovine Expression Array. Keywords: time course

ORGANISM(S): Bos taurus Ovis aries

PROVIDER: GSE5195 | GEO | 2006/09/19

SECONDARY ACCESSION(S): PRJNA96539

REPOSITORIES: GEO

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Transcription profiling of callipyge lambs at the onset of muscle hypertrophy in : time course

Project description:The callipyge mutation causes postnatal muscle hypertrophy in heterozygous lambs that inherit a paternal callipyge allele (+/CLPG). Our hypothesis was that the up-regulation of one or both of the affected paternally expressed genes (DLK1 or PEG11) initiates changes in biochemical and physiological pathways in skeletal muscle to induce hypertrophy. The goal of this study was to identify changes in gene expression during the onset of muscle hypertrophy in order to identify the pathways that are involved in the expression of the callipyge phenotype. Gene expression was analyzed in longissimus dorsi total RNA from lambs at 10, 20, and 30 days of age using the Affymetrix Bovine Expression Array. Experiment Overall Design: Longissimus dorsi were collected from 10, 20, and 30 day old callipyge and normal lambs with 2 biological replicates for each group. Total RNA was isolated and hybridized to Affymetrix Bovine Expression arrays. Data were analyzed in SAS and all genotype effects were followed up by qPCR in 42 lambs ranging from 10 to 200 days of age.

2008-11-07 | E-GEOD-5195 | biostudies-arrayexpress

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Expression data from Sheep kidney fat (KF) in lambs at 12 weeks of age; comparison of two genotypes, Callipyge (NCpat (CN)) and wild type (NN)

Project description:The ovine Callipyge mutation causes postnatal muscle hypertrophy localized to the pelvic limbs and torso, as well as body leanness. The mechanism underpinning enhanced muscle mass is unclear, as is the systemic impact of the mutation. Using muscle fibre typing immunohistochemistry we confirmed muscle specific effects and demonstrated that affected muscles had greater prevalence and hypertrophy of type 2X fast twitch glycolytic fibres and decreased representation of types 1, 2C, 2A and/or 2AX fibres. To investigate potential systemic effects of the mutation, proton NMR spectra of plasma taken from lambs at 8 and 12 weeks of age were measured. Multivariate statistical analysis of plasma metabolite profiles demonstrated effects of development and genotype but not gender. Plasma from Callipyge lambs at 12 weeks of age, but not 8 weeks, was characterized by a metabolic profile consistent with contributions from the affected hypertrophic fast twitch glycolytic muscle fibres. Microarray analysis of the perirenal adipose tissue depot did not reveal a transcriptional effect of the mutation in this tissue. We conclude that there is an indirect systemic effect of the Callipyge mutation in skeletal muscle in the form of changes of blood metabolites, which may contribute to secondary phenotypes such as body leanness. Microarrays were used for transcription profiling of kidney fat samples taken from Callipyge (n=4) and wild type (n=4) lambs at 12 weeks of age. Sheep used in this experiment were bred from a research flock of Dorset/Suffolk/Rambouillet cross-bred sheep raised at Utah State University and cared for and euthanased for sample collection in accordance with the animal ethics guidelines of Utah State University (Utah, USA). Kidney fat (KF) samples were taken from lambs at 12 weeks of age; four callipyge (CN) genotype animals and four normal (NN) genotype animals were compared.

2014-06-03 | E-GEOD-58148 | biostudies-arrayexpress

Expression data from Sheep kidney fat (KF) in lambs at 12 weeks of age; comparison of two genotypes, Callipyge (NCpat (CN)) and wild type (NN)

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Transcription profiling of lamb Callipyge muscle types (ST, LD, SS, SM) at birth (T=0) and 12 weeks (T=12)

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Assessing the effect of the CLPG mutation on the microRNA catalogue of skeletal muscle using high-throughput sequencing

Project description:The callipyge phenotype is a monogenic muscular hypertrophy that is only expressed in heterozygous sheep receiving the CLPG mutation from their sire. The wild-type phenotype of CLPG/CLPG animals is thought to result from translational inhibition of paternally expressed DLK1 transcripts by maternally expressed miRNAs. To identify the miRNA responsible for this trans-effect, we used high-throughput sequencing to exhaustively catalogue miRNAs expressed in skeletal muscle of sheep of the four CLPG genotypes. We have identified 747 miRNA species of which 110 map to the DLK1-GTL2 or callipyge domain. We demonstrate that the latter are imprinted and preferentially expressed from the maternal allele. We show that the CLPG mutation affects their level of expression in cis (~3.2-fold increase) as well as in trans (~1.8-fold increase). In CLPG/CLPG animals, miRNAs from the DLK1-GTL2 domain account for ~20% of miRNAs in skeletal muscle.We show that the CLPG genotype affects the levels of A to I editing of at least five pri-miRNAs of the DLK1-GTL2 domain, but that levels of editing of mature miRNA are always minor. We present suggestive evidence that the miRNAs from the domain target the ORF of DLK1 thereby causing the trans-inhibition underlying polar overdominance. We highlight the limitations of high-throughput sequencing for digital gene expression profiling as a result of biased and inconsistent amplification of specific miRNAs.

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