Intramuscular fat
Intramuscular fat (IMF), also known as intramuscular adipose tissue (IMAT), is white adipose tissue deposited within the skeletal muscle, specifically between bundles of muscle fibers or fascicles, distinct from intermuscular fat located between muscle groups and intramyocellular lipids stored inside muscle cells.[1] This deposition occurs through the differentiation of fibro/adipogenic progenitors (FAPs), mesenchymal cells that express platelet-derived growth factor receptor alpha (PDGFRα), into adipocytes.[2] In physiological contexts, IMF functions primarily as an energy reserve, similar to other white adipose depots, and may support muscle regeneration by aiding satellite cell function in healthy states.[3] However, its accumulation is tightly regulated by genetic factors (e.g., PPARγ and C/EBPα transcription factors), nutrition, hormones like leptin and testosterone, and physical activity, with deposition often increasing post-puberty or during high-energy diets.[1] In humans, moderate IMF contributes to metabolic homeostasis, but excessive levels—exceeding approximately 12% of muscle area—act as a physical barrier to muscle contraction and secrete pro-inflammatory adipokines (e.g., TNFα, IL-6), leading to insulin resistance and reduced muscle quality.[2] Pathologically, IMF infiltration is a hallmark of conditions including sarcopenia, obesity, muscular dystrophies (e.g., Duchenne muscular dystrophy), chronic obstructive pulmonary disease (COPD), and muscle injury, where it correlates with diminished strength, mobility impairment, and higher all-cause mortality risk independent of overall body fat.[2] Aging and inactivity promote this ectopic fat buildup via disrupted signaling pathways like Wnt and Hedgehog, while exercise and certain diets can mitigate it.[3] In livestock such as cattle and pigs, IMF—termed marbling—is economically valuable, enhancing meat tenderness, flavor, and juiciness through its even distribution and higher content of monounsaturated fatty acids, with breeds like Wagyu exhibiting up to 37.8% IMF compared to 2.8% in Brahman cattle.[1] Overall, while IMF plays beneficial roles in energy storage and tissue quality, its dysregulation underscores its dual nature as a marker of metabolic health.[3]Definition and Physiology
Composition and Location
Intramuscular fat (IMF), also known as intramuscular adipose tissue (IMAT), is white adipose tissue deposited within skeletal muscle, specifically between bundles of muscle fibers or fascicles.[1] It arises through the differentiation of fibro/adipogenic progenitors (FAPs), mesenchymal cells expressing platelet-derived growth factor receptor alpha (PDGFRα), into adipocytes that primarily store triglycerides in large lipid droplets, with minor amounts of phospholipids and cholesterol esters. This composition and location distinguish IMF from intramyocellular lipids (IMCL), which are small triglyceride-rich droplets stored within the cytoplasm of individual muscle fibers, and from intermuscular fat located between major muscle groups.[4] Anatomically, IMF occupies the extracellular space between fascicles, integrating into the muscle structure to provide localized fat storage, unlike subcutaneous fat beneath the skin or visceral fat around organs, which are larger systemic reservoirs of adipocytes. In humans, IMF distribution varies by muscle type, age, and metabolic status, but it is generally more prominent in certain muscles like the thigh and calf. The concept of intramuscular fat was initially recognized in meat science during the early 20th century, where it is termed "marbling" for the visible white fat streaks within muscle that enhance meat tenderness, flavor, and juiciness.[5] In human physiology, systematic study of IMF began in the 1990s, aided by advances in imaging and biopsy methods that clarified its patterns in healthy and diseased states.Biological Functions
Intramuscular fat serves primarily as an energy reserve, storing triglycerides that can be hydrolyzed and mobilized as fatty acids for oxidation during prolonged energy demands, akin to other white adipose depots.[3] In healthy physiological contexts, moderate IMF levels may support muscle regeneration by creating a favorable microenvironment that aids satellite cell activation and function during repair processes.[6] Beyond energy storage, IMF provides mechanical cushioning to muscle fibers, potentially mitigating shear forces during contraction and contributing to overall tissue resilience. Its accumulation and utilization are regulated by hormones such as insulin, which promotes lipogenesis and storage, and catecholamines, which stimulate lipolysis via adrenergic receptors, as well as transcription factors like PPARγ that coordinate lipid metabolism genes.[1]Measurement and Assessment
Imaging and Non-Invasive Methods
Magnetic resonance imaging (MRI) is a primary non-invasive method for quantifying intramuscular fat, particularly through the proton density fat fraction (PDFF), which measures the ratio of mobile protons from fat (triglycerides) to the total density of protons from both fat and water.[7] PDFF provides an accurate assessment of intramuscular fat (IMF) content by correcting for confounding factors like T2* relaxation and field inhomogeneities.[8] A key protocol in MRI for this purpose is the Dixon method, which exploits the chemical shift difference between water and fat signals to generate separate images and quantify fat fractions, enabling precise separation of intramuscular fat from water components in skeletal muscle.[9] Studies have validated PDFF against muscle biopsies, showing strong correlations (r > 0.9) between MRI-derived fat fractions and histologically measured fat percentages in diseased muscles.[10] Computed tomography (CT) estimates intramuscular fat infiltration using attenuation values in Hounsfield units (HU), where lower values indicate higher fat content due to the distinct density differences between fat (-190 to -30 HU) and muscle tissue (positive HU).[11] This approach allows for the calculation of fat infiltration ratios by thresholding regions of interest in muscle cross-sections, though it primarily assesses macroscopic intramuscular fat and requires careful region-of-interest selection to exclude intermuscular adipose tissue.[12] CT attenuation has been correlated with direct lipid content from biopsies, confirming its utility as a marker of myosteatosis in older adults.[11] However, repeated CT scans involve ionizing radiation, limiting its use in longitudinal studies. Ultrasound employs echo intensity (EI) analysis to assess muscle echogenicity, where increased grayscale values reflect higher intramuscular fat content due to greater sound wave reflection from lipid deposits compared to lean muscle.[13] EI measurements, often taken from the vastus lateralis or rectus femoris, correlate moderately with fat fractions derived from MRI (r ≈ 0.6-0.8), providing a bedside estimate of muscle quality.[14] This method is particularly useful for detecting age-related or disease-associated fat accumulation.[15] Each technique offers distinct advantages and faces specific limitations in clinical and research applications. MRI and CT provide high-resolution, volumetric quantification of fat distribution without operator dependency, making them gold standards for precise IMF assessment, though their high cost, limited accessibility, and need for specialized equipment restrict widespread use.[16] Ultrasound excels in portability, low cost, and real-time imaging, facilitating routine screening, but suffers from operator variability, subcutaneous fat interference, and limited penetration depth (typically <6 cm), which can affect reproducibility.[17] All methods have been validated against biopsy standards, with MRI showing the strongest direct correlations to histological fat content, while ultrasound and CT offer complementary, indirect measures.[10]Biochemical and Invasive Techniques
Biochemical and invasive techniques provide direct, high-resolution analysis of intramyocellular lipids (IMCL) and intramuscular fat (IMF), enabling quantification at the cellular and molecular levels through tissue extraction and specialized assays.[18] Muscle biopsy remains the gold standard for invasive assessment, typically involving percutaneous needle extraction from sites such as the vastus lateralis muscle to obtain small tissue samples for IMCL and IMF evaluation.[19] For IMF, histological analysis quantifies the percentage of muscle area occupied by adipocytes using stains like hematoxylin-eosin and digital image processing. Following extraction, cryosections of the biopsy are stained with Oil Red O to visualize lipid droplets, which appear as red inclusions within muscle fibers, allowing qualitative and semi-quantitative assessment of IMCL distribution.[20] For ultrastructural detail, transmission electron microscopy of biopsy samples reveals the size, number, and subsarcolemmal or intermyofibrillar location of lipid droplets, providing insights into their morphological characteristics and proximity to mitochondria.[21] Biochemical assays on homogenized biopsy tissue enable precise molecular quantification of IMCL components. Enzymatic hydrolysis methods, involving lipases to break down triglycerides into glycerol and fatty acids, are used to measure total triglyceride content, often expressed as micromoles per gram of wet tissue. Advanced techniques like mass spectrometry further profile specific lipid species, such as diacylglycerols, by identifying their molecular composition and abundance, which can indicate metabolic perturbations in muscle lipid storage.[18] In vivo tracing of IMCL dynamics employs stable isotope-labeled fatty acids, administered intravenously, to track incorporation and turnover rates within muscle lipid pools via subsequent biopsy and gas chromatography-mass spectrometry analysis.[22] This approach quantifies fatty acid flux, revealing synthesis and oxidation rates of IMCL under various physiological conditions.[23] Standardization of these techniques faces challenges from inter-individual and procedural variability, including differences in biopsy site selection—such as the vastus lateralis, where fiber type distribution can affect IMCL measurements—and processing artifacts like uneven freezing or staining inconsistencies that alter lipid droplet visibility.[24] Repeated biopsies may also induce local tissue changes, contributing to measurement variability across studies.[25] These methods often correlate with non-invasive imaging for validation but offer superior specificity for molecular profiling.Metabolic Role
Energy Utilization During Activity
Intramuscular fat (IMF), deposited as adipocytes between muscle fiber bundles, primarily serves as a long-term energy reserve within skeletal muscle, contributing to overall fat oxidation during prolonged physical activity or energy deficits rather than providing rapid fuel like intramyocellular lipids (IMCL). Unlike IMCL, which can supply up to 25% of energy expenditure during moderate-intensity endurance exercise (e.g., at 60-65% VO₂max), IMF mobilization is slower and more dependent on hormonal signals such as low insulin and high catecholamines, potentially accounting for a smaller, supportive role in total fatty acid provision during extended sessions like marathon running.[26][27] In endurance-trained individuals, chronic exercise adaptations may indirectly enhance IMF utilization by improving overall lipid metabolism and reducing IMF accumulation through mechanisms like increased Wnt signaling, which inhibits adipogenesis. However, direct measurements of IMF depletion during acute exercise are limited, with studies suggesting minimal short-term changes compared to the 20-60% IMCL reductions observed in type I fibers post-exercise.[3] This positions IMF as a structural energy depot that supports sustained activity by maintaining local fatty acid availability without significantly depleting during moderate bouts.[28] The turnover of IMF is influenced by training status and nutrition, with resynthesis occurring over days to weeks following energy demand, aiding muscle recovery and adaptation. While not the primary substrate, excessive IMF may impair energy efficiency by acting as a mechanical barrier to contraction.[1] Sex-based differences in IMF metabolism during exercise are less pronounced than for IMCL, though estrogen may promote greater overall fat utilization in females, potentially including minor contributions from IMF via enhanced lipolysis in adipose depots.[29]Interaction with Insulin Signaling
Intramuscular fat (IMF) modulates insulin signaling primarily through paracrine and endocrine effects from its adipocytes, rather than intracellular lipid intermediates like those from IMCL. In states of excess accumulation, such as obesity or sarcopenia, IMF adipocytes secrete pro-inflammatory adipokines including tumor necrosis factor-alpha (TNFα) and interleukin-6 (IL-6), which activate JNK and IKKβ pathways in adjacent muscle fibers, leading to serine phosphorylation of insulin receptor substrate-1 (IRS-1) and inhibition of tyrosine phosphorylation. This disrupts IRS-1 association with PI3K, impairing Akt activation and GLUT4 translocation, thereby reducing insulin-stimulated glucose uptake.[2][3] Additionally, IMF contributes to systemic insulin resistance by promoting free fatty acid (FFA) release into local and circulating pools, elevating plasma FFA levels that can double in obese individuals compared to lean controls and drive hepatic gluconeogenesis. This FFA spillover, arising from incomplete oxidation and adipokine-mediated lipolysis, correlates with IMF content and exacerbates hyperglycemia.[30][31] In pathological conditions, IMF infiltration physically hinders muscle function and amplifies inflammation, with studies linking higher IMF to up to 50% reductions in insulin sensitivity independent of IMCL. However, in healthy or trained states, moderate IMF may support metabolic homeostasis without impairing signaling.[32] A protective effect akin to the "athlete's paradox" for IMCL is observed with exercise, which reduces IMF via enhanced mitochondrial function and anti-inflammatory signaling, preserving insulin sensitivity despite stable or slightly elevated fat levels.[33] The pathway for IMF-induced insulin resistance can be summarized as:- IMF adipocyte secretion → Pro-inflammatory adipokines (TNFα, IL-6)
- Adipokine activation → JNK/IKKβ signaling
- Inflammatory effects → Serine phosphorylation of IRS-1, reduced tyrosine phosphorylation
- Downstream impairment → Decreased PI3K/Akt signaling → Reduced GLUT4 translocation and glucose uptake