Clinical Application of Skeletal Muscle Quantity and Quality Assessment Using Bioelectrical Impedance and Ultrasound Images.

A decline in muscle strength is a key factor responsible for physical dysfunction in older individuals. Both loss of muscle quantity and quality are associated with muscle strength decline. While the gold standard method for evaluating muscle mass and quality is magnetic resonance imaging, it is not suitable for clinical settings because of the measurement and analysis costs. Bioelectrical impedance analysis (BIA) and B-mode ultrasonography are clinically useful alternatives for skeletal muscle assessment owing to their feasibility and noninvasiveness. The recent advancements in the techniques for BIA and ultrasonography have improved their accuracy in assessing skeletal muscle quantity and quality, making them useful in detecting age-related and disease-specific alterations. This review comprehensively analyzes the advantages of using BIA and ultrasound imaging for assessing skeletal muscle quantity and quality and detecting muscle degeneration. We summarize the recent findings regarding age-related changes in muscle characteristics and the associations of muscle degeneration with physical dysfunction in patients with knee osteoarthritis. Furthermore, we discuss the clinical application of skeletal muscle assessment using BIA and ultrasound for evaluating training effects and exercise prescription.

Sex differences in quantitative ultrasonographic measurements of the rectus femoris in children.

The distribution and amount of intramuscular fat and fibrous tissue can be influenced by biological sex and impact muscle quality in both the functional (force-generating capacity) and morphological (muscle composition) domains. While ultrasonography (US) has proven effective in assessing age- or sex-related differences in muscle quality, limited information is available on sex differences in children. Quantitative ultrasonographic measurements, such as echo intensity (EI), EI bands (number of pixels across 50-unit intervals) and texture, may offer a comprehensive framework for identifying sex differences in muscle composition. The aim of our study was to examine the effect of sex on the rectus femoris (RF) muscle quality in children. We used EI (mean and bands) and texture as muscle quality estimates derived from B-mode US. We hypothesised that RF muscle quality differs significantly between girls and boys. Additionally, we also hypothesised that there is a significant correlation between EI bands and texture. Forty-four non-active healthy children were recruited (n = 22 girls, 12.8 ± 1.5 years; and n = 22 boys, 13.5 ± 1.2 years). RF was assessed using EI mean, EI bands, and texture analysis (homogeneity and correlation) using the Gray-Level Co-Occurrence Matrix. The results revealed significant (p < 0.05) sex differences in RF EI bands and texture. Boys displayed higher values in the 0-50 EI band and had more homogeneous muscle texture than girls. Conversely, girls displayed greater values in the 51-100 EI band and had less homogenous texture compared to boys (p < 0.05). A positive correlation was observed between the 0-50 EI band and muscle homogeneity. However, the 51-100 EI band correlated negatively with homogeneity (p < 0.05), particularly for girls. In conclusion, our study revealed sex-specific differences in mean EI, EI bands, and texture of the RF muscle in children. The variations in the correlations between the first and second EI bands and texture reveal different levels of homogeneity in each band. This indicates that distinct muscle tissue constituents, such as intramuscular fat and/or connective tissue, may be reflected in EI bands. Overall, the methods used in this study may be useful for examining muscle quality in healthy children and those with medical conditions.

Exploration of Genes Related to Intramuscular Fat Deposition in Xinjiang Brown Cattle.

The aim of this study was to investigate the differentially expressed genes associated with intramuscular fat deposition in the longissimus dorsi muscle of Xinjiang Brown Bulls. The longissimus dorsi muscles of 10 Xinjiang Brown Bulls were selected under the same feeding conditions. The intramuscular fat content of muscle samples was determined by the Soxhlet extraction method, for which 5 samples with high intramuscular fat content (HIMF group) and 5 samples with low intramuscular fat content (LIMF group) were selected. It was found that the intramuscular fat content of the HIMF group was 46.054% higher than that of the LIMF group. Muscle samples produced by paraffin sectioning were selected for morphological observation. It was found that the fat richness of the HIMF group was better than that of the LIMF group. Transcriptome sequencing technology was used to analyze the gene expression differences of longissimus dorsi muscle. Through in-depth analysis of the longissimus dorsi muscle by transcriptome sequencing technology, we screened a total of 165 differentially expressed genes. The results of Gene Ontology (GO) enrichment analysis showed that the differentially expressed genes in the two groups were mainly clustered in biological pathways related to carbohydrate metabolic processes, redox processes and oxidoreductase activities. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that the differentially expressed genes were significantly clustered in 15 metabolic pathways, which mainly covered fatty acid metabolism (related to lipid metabolism and glucose metabolism), the pentose phosphate pathway, the Peroxisome Proliferator-Activated Receptor (PPAR) signaling pathway and other important metabolic processes. The three genes that were predominantly enriched in the glycolipid metabolic pathway by analysis were SCD5, CPT1C and FBP2, all of which directly or indirectly affect intramuscular fat deposition. In summary, the present study investigated the differences in gene expression between high and low intramuscular fat content in the longissimus dorsi muscle of Xinjiang Brown Bulls by transcriptome sequencing technology and revealed the related signaling pathways. Therefore, we hypothesized that SCD5, CPT1C and FBP2 were the key genes responsible for the significant differences in intramuscular fat content of the longissimus dorsi muscles in a population of Xinjiang Brown Bulls. We expect that these findings will provide fundamental support for subsequent studies exploring key genes affecting fat deposition characteristics in Xinjiang Brown Bulls.

Molecular Characterizations of FAM13A and Its Functional Role in Inhibiting the Differentiation of Goat Intramuscular Adipocytes through RIG-I Receptor Signaling Pathway.

The aim of this study was to elucidate the effect of FAM13A on the differentiation of goat intramuscular precursor adipocytes and its mechanism of action. Here, we cloned the CDS region 2094 bp of the goat FAM13A gene, encoding a total of 697 amino acid residues. Functionally, overexpression of FAM13A inhibited the differentiation of goat intramuscular adipocytes with a concomitant reduction in lipid droplets, whereas interference with FAM13A expression promoted the differentiation of goat intramuscular adipocytes. To further investigate the mechanism of FAM13A inhibiting adipocyte differentiation, 104 differentially expressed genes were screened by RNA-seq, including 95 up-regulated genes and 9 down-regulated genes. KEGG analysis found that the RIG-I receptor signaling pathway, NOD receptor signaling pathway and toll-like receptor signaling pathway may affect adipogenesis. We selected the RIG-I receptor signaling pathway enriched with more differential genes as a potential adipocyte differentiation signaling pathway for verification. Convincingly, the RIG-I like receptor signaling pathway inhibitor (HY-P1934A) blocked this pathway to save the phenotype observed in intramuscular adipocyte with FAM13A overexpression. Finally, the upstream miRNA of FAM13A was predicted, and the targeted inhibition of miR-21-5p on the expression of FAM13A gene was confirmed. In this study, it was found that FAM13A inhibited the differentiation of goat intramuscular adipocytes through the RIG-I receptor signaling pathway, and the upstream miRNA of FAM13A (miR-21-5p) promoted the differentiation of goat intramuscular adipocytes. This work extends the genetic regulatory network of IMF deposits and provides theoretical support for improving human health and meat quality from the perspective of IMF deposits.

microRNA Temporal-Specific Expression Profiles Reveal longissimus dorsi Muscle Development in Tianzhu White Yak.

As a class of regulatory factors, microRNAs (miRNAs) play an important role in regulating normal muscle development and fat deposition. Muscle and adipose tissues, as major components of the animal organism, are also economically important traits in livestock production. However, the effect of miRNA expression profiles on the development of muscle and adipose tissues in yak is currently unknown. In this study, we performed RNA sequencing (RNA-Seq) on Tianzhu white yak longissimus dorsi muscle tissue obtained from calves (6 months of age, M6, n = 6) and young (30 months of age, M30, n = 6) and adult yak (54 months of age, M54, n = 6) to identify which miRNAs are differentially expressed and to investigate their temporal expression profiles, establishing a regulatory network of miRNAs associated with the development of muscle and adipose. The results showed that 1191 miRNAs and 22061 mRNAs were screened across the three stages, of which the numbers of differentially expressed miRNAs (DE miRNAs) and differentially expressed mRNAs (DE mRNAs) were 225 and 450, respectively. The expression levels of the nine DE miRNAs were confirmed using a reverse transcription quantitative PCR (RT-qPCR) assay, and the trend of the assay results was generally consistent with the trend of the transcriptome profiles. Based on the expression trend, DE miRNAs were categorized into eight different expression patterns. Regarding the expression of DE miRNAs in sub-trends Profile 1 and Profile 2 (p < 0.05), the gene expression patterns were upregulated (87 DE miRNAs). Gene ontology (GO) and Kyoto Encyclopedia of Genes Genomes (KEGG) analyses showed that the identified DE miRNAs and DE mRNAs were enriched in pathway entries associated with muscle and intramuscular fat (IMF) growth and development. On this basis, we constructed a DE miRNA-mRNA interaction network. We found that some DE mRNAs of interest overlapped with miRNA target genes, such as ACSL3, FOXO3, FBXO30, FGFBP4, TSKU, MYH10 (muscle development), ACOX1, FADS2, EIF4E2, SCD1, EL0VL5, and ACACB (intramuscular fat deposition). These results provide a valuable resource for further studies on the molecular mechanisms of muscle tissue development in yak and also lay a foundation for investigating the interactions between genes and miRNAs.

The Effects of Exercise Interventions on Ectopic and Subcutaneous Fat in Patients with Type 2 Diabetes Mellitus: A Systematic Review, Meta-Analysis, and Meta-Regression.

Background/Objectives: The aim of the present study was to determine the effects of exercise training on ectopic and subcutaneous fat in patients with type 2 diabetes mellitus (T2DM). Methods: Web of Science, PubMed, and Scopus were searched for original articles published through November 2023 that included exercise versus control interventions on body mass (BM), liver fat percentage, visceral fat area (VFA), subcutaneous fat area (SFA), and intramuscular fat volume or mass (IMF) in patients with T2DM. Weighted mean differences (WMDs) for liver fat and BM, standardized mean differences (SMDs) for VFA, SFA, and IMF, and 95% confidence intervals (95% CIs) were determined using random-effects models. Results: Thirty-six studies comprising 2110 patients with T2DM were included in the present meta-analysis. Exercise training effectively reduced BM [WMD = -2.502 kg, p = 0.001], liver fat% [WMD = -1.559%, p = 0.030], VFA [SMD = -0.510, p = 0.001], and SFA [SMD = -0.413, p = 0.001] in comparison to the control. The IMF [SMD = 0.222, p = 0.118] remained unchanged compared to the controls. Subgroup analyses showed that the type of exercise, duration, and body mass index (BMI) of participants were sources of heterogeneity. Conclusions: The current meta-analysis provides strong evidence that exercise training, particularly aerobic and combined (aerobic and resistance) exercise programs, is effective for reducing BM, VFA, and SFA in patients with T2DM. However, aerobic exercise was more effective for reducing liver fat than combined exercise. The beneficial effects of exercise on VFA and SFA reduction, but not liver fat, are associated with weight loss. These findings highlight the importance of including consistent exercise as a key management component for T2DM and associated ectopic fat deposition, with potential long-term benefits for metabolic health.

Consumer eating quality and physicochemical traits of pork Longissimus and Semimembranosus differed between genetic lines.

Pork eating quality is affected by various factors. In this study, Longissimus thoracis et lumborum (LTL) and Semimembranosus (SM) muscles from seven genetic lines (PM-LR - Pure maternal, Landrace-type; PM-LW - Pure maternal, Large White-type; PM-D - Pure maternal, Duroc-type; PT-D - Pure terminal, Duroc-type; PT-LW - Pure terminal, Large White-type; PT-LR - Pure Terminal, Landrace-type; Comp-P × LW × D - Composite Terminal - Pietran × Large white × Duroc) were analyzed for pH, intramuscular fat (IMF) content, and collagen content and solubility. A consumer sensory test using check-all-that-apply (CATA) and biometric approaches was also conducted. The results showed that the IMF content of line PM-D was the highest (P = 0.004), while line PT-LW received the highest score in tenderness, liking of flavor, purchase intent, and quality grading (P < 0.05). Line PM-LR and PT-LR showed the lowest IMF content and were least preferred by consumers. Compared to LTL, SM showed higher pH, collagen solubility, and sensory scores in tenderness, juiciness, liking of flavor, and overall liking (P < 0.05). Different muscles and lines were associated with different CATA terms but not with differences in consumer emotional responses. pH positively influenced tenderness, juiciness, and overall liking (P < 0.05), but IMF and collagen had little effect. The flavor was the most important sensory attribute contributing to overall liking, followed by tenderness. Genetic line and muscle affected pork chemical properties and eating quality. The findings are important for the Australian pork industry to improve the eating quality of their products.

Changing Australia's trading language has enhanced the implementation of objective carcase measurement technologies.

In 2016 an Australian project, the Advanced Livestock Measurement Technologies project (ALMTech), was initiated to accelerate the development and implementation of technologies that measure lean meat yield and eating quality. This led to the commercial testing, and implementation of a range of new technologies in the lamb, beef, and pork industries. For measuring lean meat yield %, these technologies included dual energy X-ray absorptiometry, hand-held microwave systems, and 3-D imaging systems. For measuring beef rib-eye traits and intramuscular fat %, both pre- and post-chilling technologies were developed. Post-chilling, a range of camera systems and near infrared spectrophotometers were developed. While pre-chilling, technologies included insertable needle probes, nuclear magnetic resonance, and X-ray systems. Initially these technologies were trained to predict the pre-existing traits already traded upon within industry. However, this approach was limiting because the technologies could measure attributes that were either non-existent in the trading language, were superior as calibrating standards, or more accurately reflected value than the pre-existing trait. Therefore, we introduced IMF% into the trading language for both beef and sheep meat, and carcase lean%, fat%, and bone% for sheep meat. These new technologies and the traits that they predict have delivered multiple benefits. Technology provider-companies are instilled with the confidence to commercialise due to the provision of achievable accreditation standards. Processors have the confidence to invest in these technologies and establish payment grids based upon their measurements. And lastly, it has enhanced data flow into genetic databases, industry data systems (MSA), and as feedback to producers.

A review of emerging technologies, nutritional practices, and management strategies to improve intramuscular fat composition in beef cattle.

The flavour, tenderness and juiciness of the beef are all impacted by the composition of the intramuscular fat (IMF), which is a key determinant of beef quality. Thus, enhancing the IMF composition of beef cattle has become a major area of research. Consequently, the aim of this paper was to provide insight and synthesis into the emerging technologies, nutritional practices and management strategies to improve IMF composition in beef cattle. This review paper examined the current knowledge of management techniques and nutritional approaches relevant to cattle farming in the beef industry. It includes a thorough investigation of animal handling, weaning age, castration, breed selection, sex determination, environmental factors, grazing methods, slaughter weight and age. Additionally, it rigorously explored dietary energy levels and optimization of fatty acid profiles, as well as the use of feed additives and hormone implant techniques with their associated regulations. The paper also delved into emerging technologies that are shaping future beef production, such as genomic selection methods, genome editing techniques, epigenomic analyses, microbiome manipulation strategies, transcriptomic profiling approaches and metabolomics analyses. In conclusion, a holistic approach combining genomic, nutritional and management strategies is imperative for achieving targeted IMF content and ensuring high-quality beef production.

Activation of skeletal carbohydrate-response element binding protein (ChREBP)-mediated de novo lipogenesis increases intramuscular fat content in chickens.

The intracellular lipids in muscle cells of farm animals play a crucial role in determining the overall intramuscular fat (IMF) content, which has a positive impact on meat quality. However, the mechanisms underlying the deposition of lipids in muscle cells of farm animals are not yet fully understood. The purpose of this study was to determine the roles of carbohydrate-response element binding protein (ChREBP) and fructose in IMF deposition of chickens. For virus-mediated ChREBP overexpression in tibialis anterior (TA) muscle of chickens, seven 5-d-old male yellow-feather chickens were used. At 10 d after virus injection, the chickens were slaughtered to obtain TA muscles for analysis. For fructose administration trial, sixty 9-wk-old male yellow-feather chickens were randomly divided into 2 groups, with 6 replicates per group and 5 chickens per replicate. The chickens were fed either a basal diet or a basal diet supplemented with 10% fructose (purity ≥ 99%). At 4 wk later, the chickens were slaughtered, and breast and thigh muscles were collected for analysis. The results showed that the skeletal ChREBP mRNA levels were positively associated with IMF content in multiple species, including the chickens, pigs, and mice (P < 0.05). ChREBP overexpression increased lipid accumulation in both muscle cells in vitro and the TA muscles of mice and chickens in vivo (P < 0.05), by activation of the de novo lipogenesis (DNL) pathway. Moreover, activation of ChREBP by dietary fructose administration also resulted in increased IMF content in mice and notably chickens (P < 0.05). Furthermore, the lipidomics analysis revealed that ChREBP activation altered the lipid composition of chicken IMF and tented to improve the flavor profile of the meat. In conclusion, this study found that ChREBP plays a pivotal role in mediating the deposition of fat in chicken muscles in response to fructose-rich diets, which provides a novel strategy for improving meat quality in the livestock industry.

Dynamic distribution of gut microbiota-metabolites during post-weaning longissimus dorsi muscle development in Ningxiang pigs.

Ningxiang pigs (NXPs) have a strong ability to deposit fat and intramuscular fat (IMF). However, microbiota-metabolite development and the role in IMF deposition have been rarely reported. Here, we compared the gut microbiota and metabolite profiles and IMF content at 30, 70, 150, 200, and 250 days of age of NXPs. The results revealed that the IMF content in NXPs increased significantly (P < 0.05) as the pigs' age extended. Additionally, the C14:0 content in the longissimus dorsi muscle at 30 and 70 days of age was significantly lower (P < 0.05) than that at 150 and 200 days of age. The Shannon index and ACE index showed a pattern of initially increasing and then decreasing. LEfSe analysis revealed that 41 differential bacteria at the genus level were specific to different growth stages, indicating the dominant bacteria's dynamic changes in the NXPs during different stages of age. Furthermore, we found that there were significant differences in cecal metabolism, the classification of differential metabolites revealed that 15.61% of compounds were fatty acyls, 13.98% were prenol lipids, and 10.57% were steroids and steroid derivatives. Next, the network analysis showed that Lachnospiraceae-XPB1014-group was positively related to 4-2-Aminophenyl-2-4-dioxobutanoic-acid, (Z)-3-Octene, 5-Methyl-furaldehyde, Propyl-2-4-decadienoate, which were also positively correlated with the IMF content. Our findings illustrated the dynamic distribution of cecal microbiota and metabolite composition at different growth stages in NXPs and their correlation with IMF deposition. These results provide a valuable insight into optimizing meat quality and overall health in post-weaning NXPs, providing a foundation for enhancement in pork product.IMPORTANCEUnderstanding the dynamic interplay between gut microbiota, metabolites, and intramuscular fat (IMF) deposition in pigs at various growth stages holds significant importance for the pork industry. This research sheds light on how the composition of gut microbiota and metabolites changes throughout the developmental stages of pigs, impacting IMF content in meat. By identifying specific bacterial genera and metabolites associated with IMF deposition, this study offers valuable insights for optimizing meat quality and health in post-weaning pigs. Such knowledge could lead to targeted interventions or management strategies aimed at enhancing pork product quality and overall profitability for producers. Ultimately, this research contributes to advancing our understanding of the complex relationship between gut microbiota, metabolites, and meat quality, offering practical implications for the swine industry.

Lysine 2-hydroxyisobutyrylation levels determined adipogenesis and fat accumulation in adipose tissue in pigs.

BACKGROUND: Excessive backfat deposition lowering carcass grade is a major concern in the pig industry, especially in most breeds of obese type pigs. The mechanisms involved in adipogenesis and fat accumulation in pigs remain unclear. Lysine 2-hydroxyisobutyrylation (Khib), is a novel protein post-translational modification (PTM), which play an important role in transcription, energy metabolism and metastasis of cancer cells, but its role in adipogenesis and fat accumulation has not been shown. RESULTS: In this study, we first analyzed the modification levels of acetylation (Kac), Khib, crotonylation (Kcr) and succinylation (Ksu) of fibro-adipogenic progenitors (FAPs), myogenic precursors (Myo) and mesenchymal stem cells (MSCs) with varied differentiation potential, and found that only Khib modification in FAPs was significantly higher than that in MSCs. Consistently, in parallel with its regulatory enzymes lysine acetyltransferase 5 (KAT5) and histone deacetylase 2 (HDAC2) protein levels, the Khib levels increased quadratically (P < 0.01) during adipogenic differentiation of FAPs. KAT5 knockdown in FAPs inhibited adipogenic differentiation, while HDAC2 knockdown enhanced adipogenic differentiation. We also demonstrated that Khib modification favored to adipogenic differentiation and fat accumulation by comparing Khib levels in FAPs and backfat tissues both derived from obese-type pigs (Laiwu pigs) and lean-type pigs (Duroc pigs), respectively. Accordingly, the expression patterns of KAT5 and HDAC2 matched well to the degree of backfat accumulation in obese- and lean-type pigs. CONCLUSIONS: From the perspective of protein translational modification, we are the first to reveal the role of Khib in adipogenesis and fat deposition in pigs, and provided new clues for the improvement of fat accumulation and distribution as expected via genetic selection and nutritional strategy in obese-type pigs.

The importance of psoas muscle on low back pain: a single-center study on lumbar spine MRI.

Background: Low back pain (LBP) is the most frequent indication to magnetic resonance imaging (MRI) examinations of the lumbosacral spine. The individual role of soft tissues, including muscles, on LBP is not fully understood and the contribution of each MRI-derived parameter of soft tissues status on the intensity of LBP has not been investigated in detail. Methods: The study design was observational retrospective, single center carried out at a University Hospital. Images were acquired using a using a 1.5 Tesla scanner. Patients completed a symptom questionnaire and rated their pain intensity using the Visual Analogue Scale (VAS). The VAS scores ​​were categorized as mild, moderate, and severe using cutoff values of 3.8 and 5.7, based on the literature. Biometric data, including weight and height, were also recorded to calculate the body mass index (BMI). The ratios between intramuscular fat infiltration and net muscle area were also calculated. Patient sample included 94 patients with LBP underwent MRI of the lumbosacral spine. Results: The stepwise analysis revealed that increasing psoas net area was associated with lower VAS levels (odds ratio [OR]: 0.94: 95% confidence interval [CI]: 0.90-0.98; p=.005), and an increase of one square centimeter of total psoas area resulted in a greater probability of reporting a mild (+1.21%; 95% CI: 0.37, 2.05%) or a moderate VAS (+0.40%; 95% CI: -0.02, 0.82%), Furthermore, a more severe VAS was associated with a higher BMI (OR: 1.13; 95% CI: 1.00-1.27). Conclusion: Our study demonstrates a relationship between LBP and MRI parameters of paravertebral and psoas muscles status. The psoas muscle is extremely important for spine stabilization and is linked to clinical symptoms of patients affected by LBP. These findings could contribute to future studies and improve treatment options in patients with LBP, possibly reducing the impact on disability, quality of life and socioeconomical burden.

Correlated Responses to Selection for Intramuscular Fat on the Gut Microbiome in Rabbits.

Intramuscular fat (IMF) content is important for meat production and human health, where the host genetics and its microbiome greatly contribute to its variation. The aim of this study is to describe the consequences of the genetic modification of IMF by selecting the taxonomic composition of the microbiome, using rabbits from the 10th generation of a divergent selection experiment for IMF (high (H) and low (L) lines differ by 3.8 standard deviations). The selection altered the composition of the gut microbiota. Correlated responses were better distinguished at the genus level (51 genera) than at the phylum level (10 phyla). The H-line was enriched in Hungateiclostridium, Limosilactobacillus, Legionella, Lysinibacillus, Phorphyromonas, Methanosphaera, Desulfovibrio, and Akkermansia, while the L-line was enriched in Escherichia, Methanobrevibacter, Fonticella, Candidatus Amulumruptor, Methanobrevibacter, Exiguobacterium, Flintibacter, and Coprococcus, among other genera with smaller line differences. A microbial biomarker generated from the abundance of four of these genera classified the lines with 78% accuracy in a logit regression. Our results demonstrate different gut microbiome compositions in hosts with divergent IMF genotypes. Furthermore, we provide a microbial biomarker to be used as an indicator of hosts genetically predisposed to accumulate muscle lipids, which opens up the opportunity for research to develop probiotics or microbiome-based breeding strategies targeting IMF.

GSK3 regulation Wnt/ß-catenin signaling affects adipogenesis in bovine skeletal muscle fibro/adipogenic progenitors.

Clarifying the cellular origin and regulatory mechanisms of intramuscular fat (IMF) deposition is crucial for improving beef quality. Here, we used single-nucleus RNA sequencing to analyze the structure and heterogeneity of skeletal muscle cell populations in different developmental stages of Yanbian cattle and identified eight cell types in two developmental stages of calves and adults. Among them, fibro/adipogenic progenitors (FAPs) expressing CD29 (ITGA7)pos and CD56 (NCAM1)neg surface markers were committed to IMF deposition in beef cattle and expressed major Wnt ligands and receptors. LY2090314/XAV-939 was used to activate/inhibit Wnt/ß-catenin signal. The results showed that the blockade of Glycogen Synthase Kinase 3 (GSK3) by LY2090314 promoted the stabilization of ß-catenin and reduced the expression of genes related adipogenic differentiation (e.g., PPARγ and C/EBPα) in bovine FAPs, confirming the anti-adipogenic effect of GSK3. XAV-939 inhibition of the Wnt/ß-catenin pathway promoted the lipid accumulation capacity of FAPs. Furthermore, we found that blocking GSK3 enhanced the paracrine effects of FAPs-MuSCs and increased myotube formation in muscle satellite cells (MuSCs). Overall, our results outline a single-cell atlas of skeletal muscle development in Yanbian cattle, revealed the role of Wnt/GSK3/ß-catenin signaling in FAPs adipogenesis, and provide a theoretical basis for further regulation of bovine IMF deposition.

Adipocyte Progenitor Pools Composition and Cellular Niches Affect Adipogenesis Divergence in Porcine Subcutaneous and Intramuscular Fat.

Intramuscular fat (IMF) contributed positively to pork quality, whereas subcutaneous fat (SCF) was often considered to be a detrimental factor impacting growth and carcass traits. Reducing SCF while maintaining optimal IMF levels requires a thorough understanding of the adipogenic differences between these two adipose depots. Our study explored the differences in adipogenesis between porcine IMF and SCF, and the results showed that subcutaneous adipocytes (SCAs) demonstrate a greater potential for adipogenic differentiation, both in vivo and in vitro. Lipidomic and transcriptomic analyses suggested that intramuscular adipocytes (IMAs) are more inclined to biosynthesize unsaturated fatty acids. Furthermore, single-cell RNA sequencing (scRNA-seq) was employed to dissect the intrinsic and microenvironmental discrepancies in adipogenesis between porcine IMF and SCF. Comparative analysis indicated that SCF was enriched with preadipocytes, exhibiting an enhanced adipogenic potential, while IMF was characterized by a higher abundance of stem cells. Furthermore, coculture analyses of porcine intramuscular adipogenic cells and myogenetic cells indicated that the niche of IMAs inhibited its adipogenic differentiation. Cell communication analysis identified 160 ligand-receptor pairs and channels between adipogenic and myogenetic cells in IMF. Collectively, our study elucidated two intrinsic and microenvironmental novel mechanisms underpinning the divergence in adipogenesis between porcine SCF and IMF.

Single-nucleus RNA sequencing and lipidomics reveal characteristics of transcriptional and lipid composition in porcine longissimus dorsi muscle.

BACKGROUND: Global per capita meat consumption continues to rise, especially pork. Meat quality is influenced by the content of intramuscular fat (IMF) as a key factor. The longissimus dorsi muscle of Dahe pigs (DHM, IMF: 7.98% ± 1.96%) and Dahe black pigs (DHBM, IMF: 3.30% ± 0.64%) was studied to explore cellular heterogeneity and differentially expressed genes (DEGs) associated with IMF deposition using single-nucleus RNA sequencing (snRNA-seq). The lipid composition was then analyzed using non-targeted lipidomics. RESULTS: A total of seven cell subpopulations were identified, including myocytes, fibroblast/fibro/adipogenic progenitors (FAPs), satellite cells, endothelial cells, macrophages, pericytes, and adipocytes. Among them, FAPs and adipocytes were more focused because they could be associated with lipid deposition. 1623 DEGs in the FAPs subpopulation of DHBM were up-regulated compared with DHM, while 1535 were down-regulated. These DEGs enriched in the glycolysis/gluconeogenesis pathway. 109 DEGs were up-regulated and 806 were down-regulated in the adipocyte subpopulation of DHBM compared with DHM, which were mainly enriched in the PPAR signaling pathway and fatty acid (FA) biosynthesis. The expression level of PPARG, ABP4, LEP, and ACSL1 genes in DHM was higher than that in DHBM. Lipidomics reveals porcine lipid composition characteristics of muscle tissue. A total of 41 lipid classes and 2699 lipid species were identified in DHM and DHBM groups. The top ten relative peak areas of lipid classes in DHM and DHBM were triglyceride (TG), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS), diglyceride (DG), cardiolipin (CL), ceramides (Cer), Simple Glc series (Hex1Cer), sphingomyelin (phSM), and phosphatidylinositol (PI). The relative peak areas of 35 lipid species in DHM were lower than DHBM, and 28 lipid species that were higher. There was a significant increase in the TG fatty acyl chains C6:0, C17:0, and C11:4, and a significant decrease in C16:0, C18:1, C18:2, and C22:4 in DHBM (p < 0.05). CONCLUSIONS: C16:0 FA may downregulate the expression level of PPARG gene, which leads to the downregulation of fat metabolism-related genes such as ACSL, PLIN2, and FABP4 in DHBM compared with DHM. This may be the reason that the lipid deposition ability of Dahe pigs is stronger than that of Dahe black pigs, which need further investigation.

Preliminary studies on the molecular mechanism of intramuscular fat deposition in the longest dorsal muscle of sheep.

BACKGROUND: Intramuscular fat content is an important index reflecting the quality of mutton, which directly affects the flavor and tenderness of mutton. Livestock and poultry intramuscular fat content is influenced by genetics, nutritional level, and environmental factors. Key regulatory factors play a crucial role in intramuscular fat deposition. However, there is a limited amount of research on the identification and function of key genes involved in intramuscular fat content deposition specifically in sheep. RESULTS: Histological differences in the longest dorsal muscle of the small-tailed frigid sheep increased in diameter and decreased in several muscle fibers with increasing monthly age; The intramuscular fat content of the longest dorsal muscle of the small-tailed cold sheep varied with age, with a minimum of 1 month of age, a maximum of 6 months of age, and a minimum of 12 months of age. Transcriptomic sequencing and bioinformatics analysis revealed a large number of differential genes in the longest dorsal muscles of little-tailed billy goats of different months of age, which were enriched in multiple GO entries and KEGG pathways. Among them, the pathway associated with intramuscular fat was the AMPK signaling pathway, and the related genes were PPARGC1A and ADIPOQ; Immunohistochemical studies showed that PPARGC1A and ADIPOQ proteins were expressed in connective tissues, cell membranes, and, to a lesser extent, the cytoplasm of the longest dorsal muscle of the little-tailed frigid sheep; Real-time PCR and Western Blot validation showed that PPARGC1A and ADIPOQ were both expressed in the longest dorsal muscle of the little-tailed frigid sheep at different ages, and there were age differences in the amount of expression. The ADIPOQ gene was negatively correlated with the intramuscular fat content of the longest dorsal muscle, and the PPARGC1A gene was positively correlated with the intramuscular fat content of the longest dorsal muscle; As inferred from the above results, the ADIPOQ gene was negatively correlated with the intramuscular fat content of the longest dorsal muscle (r = -0.793, P < 0.05); and the PPARGC1A gene was positively correlated with the intramuscular fat content of the longest dorsal muscle r = 0.923, P < 0.05). CONCLUSIONS: Based on the above results, it can be inferred that the ADIPOQ gene is negatively correlated with the intramuscular fat content of the longest back muscle (r = -0.793, P < 0.05); the PPARGC1A gene is positively correlated with the intramuscular fat content of the longest back muscle (r = 0.923, P < 0.05).

Metabolic and microbial mechanisms related to the effects of dietary wheat levels on intramuscular fat content in finishing pigs.

The current study aimed to investigate the metabolic and microbial mechanisms behind the effects of dietary wheat levels on intramuscular fat (IMF) content in the psoas major muscle (PM) of finishing pigs. Thirty-six barrows were arbitrarily assigned to two groups and fed with diets containing 25% or 55% wheat. Enhancing dietary wheat levels led to low energy states, resulting in reduced IMF content. This coincided with reduced serum glucose and low-density lipoprotein cholesterol levels. The AMP-activated protein kinase α2/sirtuin 1/peroxisome proliferator-activated receptor-γ coactivator 1α pathway may be activated by high-wheat diets, causing downregulation of adipogenesis and lipogenesis genes, and upregulation of lipolysis and gluconeogenesis genes. High-wheat diets decreased relative abundance of Lactobacillus and Coprococcus, whereas increased SMB53 proportion, subsequently decreasing colonic propionate content. Microbial glycolysis/gluconeogenesis, d-glutamine and D-glutamate metabolism, flagellar assembly, and caprolactam degradation were linked to IMF content. Metabolomic analysis indicated that enhancing dietary wheat levels promoted the protein digestion and absorption and affected amino acids and lipid metabolism. Enhancing dietary wheat levels reduced serum glucose and colonic propionate content, coupled with strengthened amino acid metabolism, contributing to the low energy states. Furthermore, alterations in microbial composition and propionate resulted from high-wheat diets were associated with primary bile acid biosynthesis, arachidonic acid metabolism, steroid hormone biosynthesis, and biosynthesis of unsaturated fatty acids, as well as IMF content. Colonic microbiota played a role in reducing IMF content through modulating the propionate-mediated peroxisome proliferators-activated receptor signaling pathway. In conclusion, body energy and gut microbiota balance collectively influenced lipid metabolism.

Transcriptome analysis reveals the effects of Schizochytrium sp. on the meat quality attributes of Tan lambs.

Schizochytrium sp., a feed additive, positively affects the quality of animal meat. In this study, the molecular mechanisms through which dietary Schizochytrium sp. affects the meat quality characteristics of Tan lambs were investigated using transcriptomic techniques. The findings demonstrate that the lambs supplemented with Schizochytrium sp. had a larger loin eye area and a higher average daily gain and intramuscular fat content (P < 0.05). They also had lower drip loss (at 24 and 48 h) and shear force (P < 0.05). Further, 745 genes were differentially expressed between lambs supplemented with Schizochytrium and the control group. Moreover, KEGG pathway analysis showed that the ECM-receptor interaction pathway, which is related to muscle generation and intramuscular fat deposition, was significantly enriched in the lambs administered a diet containing Schizochytrium sp. Herein, we identified some pivotal genes linked to muscular system development and lipid metabolism. Thus, using Schizochytrium sp. may boost the meat quality of Tan lambs by modifying the expression of genes related to hub pathways. The results supply a new basis to determine the molecular mechanisms through which Schizochytrium sp. supplementation regulates the meat quality characteristics of sheep.