We examined the ability of n-3 FA in flaxseed-supplemented rations to increase the n-3 FA content of bovine muscle. Two groups of animals were used in each of two separate trials: (i) Hereford steers supplemented (or not) with ground flaxseed (907 g/d) for 71 d, and (ii) Angus steers supplemented (or not) with ground flaxseed (454 g/d for 3 d followed by 907 g/d for 110 d). For the Hereford group, flaxseed-supplemented rations increased 18:3n-3 (4.0-fold), 20:5n-3 (1.4-fold), and 22:5n-3 (1.3-fold) mass as compared with the control, and increased total n-3 mass about 1.7-fold. When these data were expressed as mol%, the increase in 18:3n-3 was 3.3-fold and in 20:5n-3 was 1.3-fold in the phospholipid fraction, and 18:3n-3 was increased 4-fold in the neutral lipid fraction. For the Angus group, flaxseed ingestion increased masses and composition of n-3 FA similarly to that for the Herefords and doubled the total n-3 FA mass. The effect of cooking to a common degree of doneness on FA composition was determined using steaks from a third group of cattle, which were Angus steers. We demonstrated no adverse effects on FA composition by grilling steaks to an internal temperature of 64 degrees C. Because n-3 FA may affect gene expression, we used quantitative real-time reverse transcriptase-polymerase chain reaction to quantify the effect of feeding flaxseed on heart-FA binding protein, peroxisome proliferator activated receptor gamma (PPARgamma) and alpha (PPARalpha) gene expression in the muscle tissue. PPARgamma mRNA level was increased 2.7-fold in the flaxseed-fed Angus steers compared with the control. Thus, we demonstrate a significant increase in n-3 FA levels in bovine muscle from cattle fed rations supplemented with flaxseed and increased expression of genes that regulate lipid metabolism.
Heart fatty acid binding protein (H-FABP) has been associated with intramuscular fat content in pigs. In the current study, we showed that expression of H-FABP mRNA in adipose tissue of adult pigs was 8.5% of that in heart and 30% of that in skeletal muscle, and that H-FABP mRNA level was more than 10% of that of adipocyte fatty acid binding protein mRNA in adipose tissue. Levels of H-FABP mRNA reached a maximum in adipose tissue from 7-d neonates, with no further increase in the adult. Also, H-FABP mRNA was induced during adipogenic differentiation of stromal-vascular cells derived from adipose tissue and skeletal muscle. In conclusion, H-FABP may play a role in adipose tissue development and function in the pig.
Adipose triglyceride lipase (ATGL), a newly identified lipase, is a rate-limiting enzyme for triglyceride hydrolysis in adipocytes. The regulatory proteins involved in ATGL-mediated lipolysis in fat tissue are not fully identified and understood. The G(0)/G(1) switch gene 2 (G0S2) is an inhibitor of ATGL activity by interacting with ATGL through the hydrophobic domain of G0S2. Here, for the first time, we have cloned the coding sequence of G0S2 cDNA for the chicken, turkey, and quail. Sequence comparisons with mammals revealed that the avian G0S2 also have a conserved hydrophobic domain. Avian G0S2 is predominantly expressed in adipose tissues relative to other tested tissues. Within the adipose tissue, G0S2 is expressed 20-fold greater in the adipocyte than in the stromal-vascular (SV) fraction (P < 0.001). Expression of G0S2 mRNA gradually increased during differentiation of chicken adipocytes in culture (P < 0.05). However, there is G0S2 expression in embryonic adipose tissue, SV fraction, and primary preadipocytes before confluence that generally have an increased capacity of cell proliferation, which indicates it has an important role in adipocyte differentiation rather than proliferation. For a better understanding of how G0S2 responds to environmental stimuli, chickens were fasted for 24 h and then refed. Expression of G0S2 in adipose tissue was dramatically decreased (P < 0.05) in the chickens and quail after a 24-h fasting period, and increased to the control level after refeeding. In contrast to G0S2 expression, ATGL expression was induced (P < 0.05) after the 24-h fasting period and rapidly returned to the control level during the refeeding period. These data indicate that changes in lipolytic activities of adipose tissue in vivo can be regulated by G0S2 expression, as an inhibitor of ATGL.
The objective of this research was to further characterize the promoter regions of the bovine and porcine fatty acid-binding protein 4 (FABP4) genes relative to those of other mammals. The DNA sequences of FABP4 promoters for the human, mouse, cow, pig, and dog were obtained from the genomic database of the National Center for Biotechnology Information and also from the sequencing of bovine and porcine genomic DNA clones obtained by 5' PCR racing of genomic DNA. Sequence alignments of these FABP4 promoters using the basic local alignment search tool of the National Center for Biotechnology Information revealed 3 highly conserved promoter regions across the species. Two computational bioinformatics databases and the literature identified the conserved transcription factor-binding sites for C/EBP, adapter primer-1, and boxes of CAAT and TATA in the first conserved proximal promoter region, a direct repeat 1-type PPAR responsive element in the second distal conserved region, and another PPAR responsive element in the third distal conserved promoter region of FABP4 in all 5 mammals. Five new short interspersed repetitive elements (SINE) in the bovine FABP4 promoter and 2 new SINE in the porcine were found, but these SINE did not disrupt the 3 conserved regions, indicating that important regulatory elements are maintained regardless of evolutionary pressure. In conclusion, the conserved cis-acting elements, especially the 2 key adipocyte transcription factors C/EBP and PPAR, may contribute greatly to adipogenic regulation and adipose tissue-specific expression of FABP4 in these mammals. This helps to further characterize and decipher important cis-acting elements that are important for adipocyte development in adipose and muscle tissue.
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