Puberty with relevance to growth and metabolism, two

Topic: HistoryCanada
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Last updated: August 25, 2019

Puberty is acomplex developmental process when animals mature into a grown adult able tosexually reproduce and is regulated by interacting genes. To enhance ourunderstanding of the relatedness between muscular growth and development andthe pubertal process, we characterised the transcriptome of the longissimus dorsi muscle of 11 heifers. Muscletissue samples from five pre- and six post- pubertal Brahman heifers wereanalysed. The expression of 14,376 was detected and 431 genes were differentiallyexpressed (DE) when pre and post heifers were compared using linear mixedmodels.

Functional enrichment analysis was carried using the list of DE genesas a target list, compared with a background list (all genes expressed ordefault for Bos Taurus species). There were XX gene ontology terms that weresignificant… and one pathway….IntroductionPuberty is a developmental process when adolescents reach sexual maturity and becomecapable of reproduction and is affected by the interaction of variousgenes. Gene interactions associated with puberty are described as gene regulatorynetwork, which is a collection of molecular regulators that interact with eachother to govern gene expression levels of RNA and proteins. Brahman heifers (Bos indicus) are cattle that originate from mostly India andSoutheast Asia and because of the heat and disease resistance, Bos indicus breeds have been usedintensively in tropical regions worldwide.

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 Age of puberty is heritable but generally in cattle, puberty occurs atabout 10 months of age.  Breeding cattlefor earlier onset of puberty through selective breeding will help improveefficiency of beef production. Muscle (Longissimus dorsi) is a tissue with relevance to growthand metabolism, two physiological processes influenced by puberty.Muscletissue is vital in heifers as it plays a part in blood circulation, breathingand digesting of the food. Without muscle tissues, heifer would even find ithard to move around. Longissimus dorsi is the spinal muscle located at the back, as it provides extension andunilterally lateralflexion of the back as it is bilaterally contracted. Muscleweight as per cent of total muscle weight in 500lb carcass (Butterfield &May, 1966) for longissimus dorsi is6.

55. In thiscurrent study, we have characterised the transcriptome of the periphery muscletissues (longissimus dorsi), known tobe related to growth and metabolism and is needed for cattle’s to reachpuberty. The tissues were collected from the pre- and post- pubertal Brahmanheifers from the same age group.

Next, the muscle tissue was prepared andquantified. Functional enrichment analysis was also done using the DE genes andthen compared with GWAS results. By the end of this study, we would be able toidentify the gene ontology and the pathways related to puberty.  Materials& Methods Retrieving of Animal Samples Heifers werehandled and managed as per approval by the Animal Ethics Committee of theUniversity of Queensland, Production and Companion Animal Group (certificatenumber QAAFI/279/12). 12 Brahman heifers were selected in this study with thesame age group and phenotypic characteristics of Bos Indicus. The heifersobserved was weighted between 176 and 224 kg.

Heifers were similarly managedand grazed a pasture-based diet at the Gatton Campus beef facilities of theUniversity of Queensland (Gatton, Australia). Observation of pubertaldevelopment was done, and the purpose was to collect tissue samples from 6 pre-and 6 post- pubertal heifers. Tissue Sample CollectionBetween the12th and 13th rib, a 1cm length of Longissimus dorsi muscle (LDM) was collected from the abdominalcavity was collected from both pre- and post- heifers. The 12 tissues werecollected within 15 minutes of post mortem, snap-frozen in liquid nitrogen andthen stored under -800C until it needs to be processed. Duringtissue processing, we failed to obtain 1 tissue sample. Thus, a total of 11tissue samples were obtainable for RNA-Seq analysis. Protein Sample Preparation for MS Individual fractions were added tolysis buffer (7M urea, 2M thiourea, 4% SDS, 10mM DTT and 1 Mm PMSF), sonicatedat power 4 for 10 seconds and vortexed vigorously at 300C for 1hour. Excess acrylamide was quenched in the samples by adding 5mM DTT, and the11 samples were precipitated with 4 volumes of methanol: acetone (1:1) at -200Covernight.

After dissolving the precipitate in 50Mm ammonium acetate, theprotein concentration was measured using Nanodrop (Thermo Scientific). 50 ?L of protein was diluted to a final volume of 100 ?Land digested by addition of 1 ?g of trypsin (Sigma) overnight at 370C. Sampleswere then desalted by C-18 Zip-tip (adapted from Millipore procedure) andanalysed by liquid chromatography electrospray ionisation tandem massspectrometry (LC-ESI-MS). Results14,376 geneswere expressed in the muscle.

431 of the pre- and post- puberty genes weredifferentially expressed, which makes it about 3%. Functional annotationbioinformatics microarray analysis was done using DAVID.  

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