A semi-quantitative RT-PCR method to measure thein vivo effect of dietary conjugated linoleic acid on porcine muscle PPAR gene expressionReportar como inadecuado




A semi-quantitative RT-PCR method to measure thein vivo effect of dietary conjugated linoleic acid on porcine muscle PPAR gene expression - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

Biological Procedures Online

, Volume 5, Issue 1, pp 20–28

Received: 01 November 2002Revised: 16 December 2002Accepted: 09 January 2003DOI: 10.1251-bpo43

Cite this article as: Meadus, W.J. Biol. Proced. Online 2003 5: 20. doi:10.1251-bpo43

Abstract

Conjugated linoleic acid CLA can activate in vitro the nuclear transcription factors known as the peroxisome proliferators activated receptors PPAR. CLA was fed at 11 g CLA-kg of feed for 45d to castrated male pigs barrows to better understand long term effects of PPAR activationin vivo. The barrows fed CLA had lean muscle increased by 3.5% and overall fat reduced by 9.2% but intramuscular fat IMF % was increased by 14% P<0.05. To measure the effect of long term feeding of CLA on porcine muscle gene expression, a semi-quantitative RT-PCR method was developed using cDNA normalized against the housekeeping genes cyclophilin and β-actin. This method does not require radioactivity or expensive PCR instruments with real-time fluorescent detection. PPAR and the PPAR responsive gene AFABP but not PPARα were significantly increased P<0.05 in the CLA fed pig’s muscle. PPARα and PPARγ were also quantitatively tested for large differences in gene expression by western blot analysis but no significant difference was detected at this level. Although large differences in gene expression of the PPAR transcriptional factors could not be confirmed by western blotting techniques. The increased expression of AFABP gene, which is responsive to PPAR transcriptional factors, confirmed that dietary CLA can induce a detectable increase in basal PPAR transcriptional activity in the live animal.

Indexing termsRT-PCR PPAR gene expression in vivo pigs muscle Published: February 17, 2003

Download to read the full article text



Autor: W. J. Meadus

Fuente: https://link.springer.com/







Documentos relacionados