Cellular and molecular characterisation of porcine congenital splayleg and the involvement of P311 and SPARCL-1 as candidate genes /
Porcine congenital splayleg (PCS) is the most important congenital condition of piglet, assciated with lameness and immbolity, of unknown actilogy and pathogenesis. the aim of this study is to investigate the cellular and molecular changes in skeletal muscles of PCS, thereby gaining new molecular in...
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| Format: | Thesis Book |
| Language: | English |
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2007.
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| 100 | 1 | |a Ooi, Peck Toung. | |
| 245 | 1 | 0 | |a Cellular and molecular characterisation of porcine congenital splayleg and the involvement of P311 and SPARCL-1 as candidate genes / |c by Ooi Peck Toung. |
| 260 | |a 2007. | ||
| 300 | |a 224 leaves : |b ill. ; |c 30cm. | ||
| 502 | |a Thesis (PhD) - University of Glasgow, 2007. | ||
| 520 | |a Porcine congenital splayleg (PCS) is the most important congenital condition of piglet, assciated with lameness and immbolity, of unknown actilogy and pathogenesis. the aim of this study is to investigate the cellular and molecular changes in skeletal muscles of PCS, thereby gaining new molecular insights into this clinical condition. Based on immunohistochemistry and histological image analyses on 4 sets of 2-day-old splayleg piglets, each with a corresponding normal litter mate, a consistent discovery has been that PCS muscles [semitendinosus (ST), longissimus dorsi (LD) and gastrocnemius (G)] showed extensive fibre atrophy without apparent tissue damage. At present, it is not certain if PCS-associated fibre atrophy is accompanied by fibre hypoplasia. Both normal and PCS muscle fibres showed similar widespread distribution of lipid-and oxidative-positive fibres. Although there was no significant difference in fibre type composition, several structural myosin heavy chain (MyHC) genes were significantly down-regulated in PCS affected muscles. Interestingly, MAFx, a major atrophy marker, was highly up-regulated in almost all PCS muscles, when highly up-regulated in almost all PCS muscles, when compared with controls from normal litter mates. In contrast, P311, a novel 8-kDa protein, was relatively down-regulated in all PCS mescles examined. To futher investigate the functional role of 311 in skeletal muscle, its full-length cDNA was sequence (Accession No. EF416570) and over-expressed in murine C2C12 mescle cells. P311 over-expression enhanced cell proliferation and reduced myotube formation in C2C12 cells. the over-expression of calcineurin, a key intracellular calcium-dependent signalling factor of muscle differentiation, down-regulated P311 expression. Reduced P311 expression in PCS piglets might contribute to atrophy through reduced myotube contribution. To investigate the functional role of SPARCL-1, a matricellular secreted glycoprotein that belongs to SPARC family, its full-length cDNA was sequenced (Accession No. EF416571) and over-expressed in murine C2C12 muscle cells. SPARCL-1 over-expression led to reduced cell proliferation and down-regulation of MyHC genesduring late differentiation. SPARCL-1 might be associated as a negative regulator of skeletal muscle proliferation and cell differentiation. However, endogenous SPARCL-1 expression was similar between PCS and normal muscles. Hence, although SPARCL-1 could play a role in muscle development, it is unlikely to be a main factor in the development of PCS. In summary, PCS is shown to be a condition characterised by extensive fibre atrophy and raised fibre density, and it is proposed that the combined differential expression of MAFbx and P311 is of potential value in the diagnosis of sub-clinical PCS. | ||
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