Florence Gondret

Intramuscular fat content in meat-producing animals: development, genetic and nutritional control, and identification of putative markers.

Intramuscular fat (IMF) content plays a key role in various quality traits of meat. IMF content varies between species, between breeds and between muscle types in the same breed. Other factors are involved in the variation of IMF content in animals, including gender, age and feeding. Variability in IMF content is mainly linked to the number and size of intramuscular adipocytes. The accretion rate of IMF depends on the muscle growth rate. For instance, animals having a high muscularity with a high glycolytic activity display a reduced development of IMF. This suggests that muscle cells and adipocytes interplay during growth. In addition, early events that influence adipogenesis inside the muscle (i.e proliferation and differentiation of adipose cells, the connective structure embedding adipocytes) might be involved in interindividual differences in IMF content. Increasing muscularity will also dilute the final fat content of muscle. At the metabolic level, IMF content results from the balance between uptake, synthesis and degradation of triacylglycerols, which involve many metabolic pathways in both adipocytes and myofibres. Various experiments revealed an association between IMF level and the muscle content in adipocyte-type fatty acid-binding protein, the activities of oxidative enzymes, or the delta-6-desaturase level; however, other studies failed to confirm such relationships. This might be due to the importance of fatty acid fluxes that is likely to be responsible for variability in IMF content during the postnatal period rather than the control of one single pathway. This is evident in the muscle of most fish species in which triacylglycerol synthesis is almost zero. Genetic approaches for increasing IMF have been focused on live animal ultrasound to derive estimated breeding values. More recently, efforts have concentrated on discovering DNA markers that change the distribution of fat in the body (i.e. towards IMF at the expense of the carcass fatness). Thanks to the exhaustive nature of genomics (transcriptomics and proteomics), our knowledge on fat accumulation in muscles is now being underpinned. Metabolic specificities of intramuscular adipocytes have also been demonstrated, as compared to other depots. Nutritional manipulation of IMF independently from body fat depots has proved to be more difficult to achieve than genetic strategies to have lipid deposition dependent of adipose tissue location. In addition, the biological mechanisms that explain the variability of IMF content differ between genetic and nutritional factors. The nutritional regulation of IMF also differs between ruminants, monogastrics and fish due to their digestive and nutritional particularities.

Comparison of intramuscular adipose tissue cellularity in muscles differing in their lipid content and fibre type composition during rabbit growth

Abstract The age-related patterns of intramuscular lipid and adipocyte characteristics in longissimus lumborum (LL) and biceps femoris (BF), two fast-twitch glycolytic muscles, and semimembranosus proprius (SMP), a slow-twitch oxidative muscle, were compared during rabbit postnatal growth. Rabbits were slaughtered at 2, 4, 5, 6, 9, 11, 14, 17 or 20 weeks of age (n=5 in each age group). In the three muscles, an elevated rate of lipid deposition occurred around week 14 of age. This increase was associated with changes in triglyceride content during the postweaning period, while the phospholipid content remained nearly constant. In SMP and LL muscles, the age-related increase in total lipid and triglyceride contents was closely associated with the increase in number and size of clustered adipocytes. Throughout the postnatal period, the slow-twitch oxidative SMP muscle displayed higher total lipid, triglyceride and phospholipid contents than the predominantly fast-twitch glycolytic LL and BF muscles. The higher lipid content of SMP was mostly the result of a much higher number of intramuscular adipocytes and, to a minor extent, of higher phospholipid and triglyceride contents within the myofibres.

Regional differences in porcine adipocytes isolated from skeletal muscle and adipose tissues as identified by a proteomic approach

The content and distribution of body lipids are of special interest for production efficiency and meat quality in the farm animal industry. Triglycerides represent the most variable fraction of tissue lipids, and are mainly stored in adipocytes. Although several studies have reported regional differences in the expression of genes and their products in adipocytes from various species, the characteristics of i.m. adipocytes remain poorly described. To evaluate adipocyte features according to muscle and other fat locations, adipocyte proteins were isolated from trapezius skeletal muscle, and intermuscular, s.c., or perirenal adipose tissues from 6 female pigs (80 d of age). Protein extracts were labeled and analyzed by 2-dimensional, fluorescent, differential gel electrophoresis. The comparisons revealed that 149 spots were always differentially expressed (P < 0.05, ratio exceeding |2|-fold difference) between i.m. adipocytes and the fat cells derived from the 3 other adipose locations. The proteins that were downregulated in i.m. fat cells belonged to various metabolic pathways, such as lipogenesis (cytosolic malate dehydrogenase and isocitrate dehydrogenase, P < 0.01), glycolysis (enolases and aldolase, P </= 0.01), lipolysis (perilipin, P < 0.01), fatty acid oxidation (long-chain fatty-acyl CoA dehydrogenase, P < 0.01), and energy transfer (catalase, voltage-dependent anion channel 1, and electron-transfer flavoprotein, P < 0.05). In contrast, both prohibitin-1 and cell division cycle 42 homolog, with possible roles in cell growth, were up-regulated (P < 0.05) in i.m. adipocytes compared with other fat cells. Fewer differences were observed when adipocytes isolated from s.c., perirenal, and intermuscular fat tissues were compared, with a maximum of 17 spots differing significantly in abundance between perirenal and s.c. adipose tissues. The findings that proteins involved in both anabolic and energy-yielding catabolic pathways are downregulated in i.m. adipocytes compared with s.c., visceral, or intermuscular adipocytes, suggest that the metabolic activity of i.m. adipocytes is low. Thus, triggering adipogenesis rather than cell metabolism per se might be a valuable strategy to control lipid deposition in pig skeletal muscles.

Differentially-expressed genes in pig Longissimus muscles with contrasting levels of fat, as identified by combined transcriptomic, reverse transcription PCR, and proteomic analyses.

Intramuscular fat content is important for many meat quality parameters. This work is aimed at identifying functional categories of genes associated with natural variation among individuals in intramuscular fat content to help the design of genetic schemes for high marbling potential. Taking advantage of the global nature of transcriptomic and proteomic technologies, 40 genes were identified as differently expressed between high fat and low fat pig Longissimus muscles at slaughter weight. They are involved in metabolic processes, cell communication, binding, and response to stimulus. Using real-time PCR in muscle biopsies taken earlier in the fattening period, the group with a high intramuscular fat content was also characterized by the down-expression of genes playing a negative role in adipogenesis, such as architectural transcription factor high-motility hook A1, mitogen activated protein-kinase14, and cyclin D1. These results suggest that interindividual variability in intramuscular fat content might arise essentially from differences in early adipogenesis.

Divergent selection on 63-day body weight in the rabbit: response on growth, carcass and muscle traits

The effects of selection for growth rate on weights and qualitative carcass and muscle traits were assessed by comparing two lines selected for live body weight at 63 days of age and a cryopreserved control population raised contemporaneously with generation 5 selected rabbits. The animals were divergently selected for five generations for either a high (H line) or a low (L line) body weight, based on their BLUP breeding value. Heritability (h2) was 0.22 for 63-d body weight (N = 4754). Growth performance and quantitative carcass traits in the C group were intermediate between the H and L lines (N = 390). Perirenal fat proportion (h2 = 0.64) and dressing out percentage (h2 = 0.55) ranked in the order L < H = C (from high to low). The weight and cross-sectional area of the Semitendinosus muscle, and the mean diameter of the constitutive myofibres were reduced in the L line only (N = 140). In the Longissimus muscle (N = 180), the ultimate pH (h2 = 0.16) and the maximum shear force reached in the Warner-Braztler test (h2 = 0.57) were slightly modified by selection.

Metabolic changes and tissue responses to selection on residual feed intake in growing pigs.

Previous selection experiments using residual feed intake (RFI) to select pigs with a high feed efficiency have reported that a low RFI was associated with a reduced body fat content and a greater muscle glycogen content. In the current study, growing Large White female piglets from 2 lines divergently selected for RFI were used to determine the changes in energy and protein metabolisms in key tissues and their cross talks in response to selection. Pigs of low RFI (RFI(-); n = 26) or high RFI (RFI(+); n = 36) selection lines were offered free access to feed during postweaning and growing periods. Pigs of each line were then slaughtered at 19 kg (n = 8 per line) or 115 kg BW (n = 14 to 18 per line). A third group of pigs of the RFI(+) line was offered feed at the same level per metabolic BW (BW0.60) as RFI- pigs (group RFI+R, n = 14). Regardless of the growth period considered, G:F was less in RFI(+) pigs than in RFI(-) pigs. At 19 kg BW, RFI(+) and RFI(-) pigs had a similar body composition and tissue lipid content. The fractional rate of protein synthesis and proteasome activity were decreased (P < 0.090) in the livers of RFI(+) pigs compared with RFI(-) pigs whereas activities of energy catabolic enzymes did not differ in the liver and LM samples. Plasma insulin was conversely greater (P = 0.049) in RFI(+) pigs at this stage. At 115 kg BW, enzyme activities of protein catabolism in the liver and in the LM did not differ (P > 0.10) between RFI(+) pigs and RFI(-) pigs. Both lactate dehydrogenase activity participating in glucose metabolism and hydroxylacylCoA dehydrogenase activity involved in fatty acid oxidation were greater (P < 0.05) in the liver and LM of RFI(+) pigs compared with RFI(-) pigs. In the liver, contrary to the LM, those differences in enzyme activities were directly associated with selection on RFI regardless of ADFI. Increased backfat depth and content and greater lipid content and adipocyte hypertrophy (P < 0.05) in subcutaneous adipose tissue were reported in RFI(+) pigs compared with RFI(-) pigs at 115 kg BW without marked changes in key lipogenic enzyme activities; these changes were directly associated with ADFI. In conclusion, the present study shows an increase of catabolic pathway activities in the liver and muscle of RFI(+) pigs at market weight that is likely to generate more ATP compared with RFI(-) pigs.

Myosin isoform transitions in four rabbit muscles during postnatal growth.

SummaryFour rabbit muscles (i.e. semimembranosus proprius, psoas major, biceps femoris and longissimus lumborum), differing in their fibre type composition in the adult, were investigated during postnatal development. Muscle samples were taken at 1, 7, 14, 21, 28, 35, 49 and 77 days of age. Complementary techniques were used to characterize myosin heavy chain (MHC) isoform transitions, i.e. SDS-PAGE, immunocytochemistry and conventional histochemistry. Good accordance was found between electrophoretic and immunocytochemical techniques. Our results show that rabbit muscles were phenotypically immature at birth. At 1 day of age, perinatal isoform represented 70–90% of the total isoform content of the muscles. Two generations of myofibres could be observed on the basis of their morphology and reaction to specific antibodies. In all muscles, primary fibres expressed slow MHC. In contrast, secondary generation of fibres never expressed slow MHC in future fast muscles, while half of them expressed slow MHC in the future slow-twitch muscle, the semimembranosus proprius. During the postnatal period, all muscles displayed a transition from embryonic to perinatal MHC isoforms, followed by a transition from perinatal to adult MHC isoforms. These transitions occured mainly during the first postnatal month. The embryonic isoform was no longer expressed after 14 days, except in longissimus where it disappeared after 28 days. On the contrary, large differences were found in the timing of disappearance of the perinatal isoform between the four muscles. The perinatal isoform disappeared between 28 and 35 days in semimembranosus proprius and 35 and 49 days in psoas and biceps femoris. Interestingly, the perinatal isoform was still present in 6% of the fibres in longissimus at 77 days, the commercial slaughter age, denoting a great delay in the maturation. Fate of each generation of fibres differed between muscles.

Prenatal exposure to maternal low or high protein diets induces modest changes in the adipose tissue proteome of newborn piglets.

The possibility that maternal diets during gestation could affect growth and tissue development of offspring and program their later phenotype is an emerging challenge in pig production. The objective of the current study was to investigate the effects of contrasted protein levels in diets of pregnant sows on the proteomic features of subcutaneous adipose tissue (SCAT) of the offspring at birth and its possible persistence later in age. Sows were fed control (Con), low (LP), or high protein (HP) diets throughout gestation. A subset of piglets was killed at 1 d of age for SCAT sampling. The remaining piglets were cross-fostered to nonexperimental sows during lactation. They were fed standard diets during postweaning and fattening periods until 186 d of age. Modifications in SCAT protein abundance shortly after birth were investigated by 2-dimensional gel electrophoresis followed by mass spectrometry. A total of 65 spots were found differentially expressed (P <or= 0.10) in SCAT of 1-d-old experimental piglets vs. Con piglets. Proteins with a greater abundance in LP piglets compared with Con piglets were involved in pathways related to glucose and fatty acid metabolisms, lipid transport, and regulation of apoptosis. Upregulation of 5 proteins representative of these biological pathways in LP group vs. Con group were further validated (P < 0.05) by Western blot analyses. Furthermore, the specific activity of the key lipogenic enzyme fatty acid synthase was found greater (P = 0.06) in SCAT of 1-d-old LP piglets than in Con piglets. The main changes evidenced in SCAT of HP piglets compared with Con animals at 1 d of age rather concerned proteins putatively involved in AA metabolism or in protein turnover. Adipose tissue contents in some proteins that had displayed a greater (P <or= 0.10) abundance in experimental pigs compared with Con at d 1 (e.g., transaldolase, annexin II, and apolipoprotein A4) were, however, similar (P > 0.10) in the 3 groups at d 186 of age. Enolase 1 has less abundance (P < 0.05) in LP pigs compared with Con pigs at this stage. In conclusion, the proteomics tool has allowed the identification of early changes in various molecular pathways of SCAT in response to the levels of maternal protein supply during gestation.

Effect of age at slaughter on chemical traits and sensory quality of Longissimus lumborum muscle in the rabbit

The effect of age on chemical composition and sensory quality of longissimus lumborum muscle was investigated in rabbits slaughtered at 11 weeks or 18 weeks of age. Intramuscular fat (IMF) content increased from 1.3 to 2.2%, crude protein content ranged from 23.4-26.9%, whereas water content declined from 74.0 to 69.3%, as the age increased. The sensory quality was assessed by a 10-member experienced sensory panel. The age was found to affect meat tenderness, whereas juiciness and flavour did not differ significantly between ages. A positive trend was observed between tenderness and IMF content, both within age and between ages. Therefore, differences in sensory quality of rabbit meat were at least partly related to IMF content.

The Loss of Adipokine Genes in the Chicken Genome and Implications for Insulin Metabolism

Gene loss is one of the main drivers in the evolution of genomes and species. The demonstration that a gene has been lost by pseudogenization is truly complete when one finds the pseudogene in the orthologous genomic region with respect to active genes in other species. In some cases, the identification of such orthologous loci is not possible because of chromosomal rearrangements or if the gene of interest has not yet been sequenced. This question is particularly important in the case of birds because the genomes of avian species possess only about 15,000 predicted genes, in comparison with 20,000 in mammals. Yet, gene loss raises the question of which functions are affected by the changes in gene counts. We describe a systematic approach that makes it possible to demonstrate gene loss in the chicken genome even if a pseudogene has not been found. By using phylogenetic and synteny analysis in vertebrates, genome-wide comparisons between the chicken genome and expressed sequence tags, RNAseq data analysis, statistical analysis of the chicken genome, and radiation hybrid mapping, we show that resistin, TNFα, and PAI-1 (SERPINE1), three genes encoding adipokines inhibiting insulin sensitivity, have been lost in chicken and zebra finch genomes. Moreover, omentin, a gene encoding an adipokine that enhances insulin sensitivity, has also been lost in the chicken genome. Overall, only one adipokine inhibiting insulin sensitivity and five adipokines enhancing insulin sensitivity are still present in the chicken genome. These genetic differences between mammals and chicken, given the functions of the genes in mammals, would have dramatic consequences on chicken endocrinology, leading to novel equilibriums especially in the regulation of energy metabolism, insulin sensitivity, as well as appetite and reproduction.