The unique physiological features of the broiler pectoralis major muscle as suggested by the three-dimensional ultrastructural study of mitochondria in type IIb muscle fibers

Typical skeletal muscles are composed of mixed muscle fiber types, which are classified as slow-twitch (type I) and fast-twitch (type II) fibers, whereas pectoralis major muscles (PMs) in broiler chickens are 100% composed of type IIb fast-twitch fibers. Since metabolic properties differ among muscle fiber types, the combination of muscle fiber types is involved in physiological functions and pathological conditions in skeletal muscles. In this study, using serial block-face scanning electron microscopy, we compared three-dimensional (3D) mitochondrial properties in type IIb fibers in broiler PMs and those in type I fibers of broiler gastrocnemius muscles (GMs) heterogeneously composed of slow- and fast-twitch muscle fibers. In type I fibers in the GMs, elongated mitochondria with numerous interconnections to form a substantial network among myofibrils were observed. Along with lipid droplets sandwiched by mitochondria, these features are an adaptation to effective oxidative respiration and constant oxidative damage in slow-twitch muscle fibers. In contrast, type IIb fibers in the PMs showed small and ellipsoid-shaped mitochondria with few interconnections and no lipid droplets, forming a sparse network. The mitochondrial spatial network comprises of active mitochondrial dynamics to reduce mitochondrial damage; therefore, type IIb fibers possess physiologically low capacity to maintain mitochondrial wellness due to static mitochondrial dynamics. Based on 3D mitochondrial properties, we discussed the contrasting physiological functions between type I and IIb fibers and proposed a high contractile power and low stress resistance as unique physiological properties of broiler PMs.     

Adaptaciones en la composici6n fibrilar del musculo Longisimo lumbar a entrenamientos de resistencia

The effect of exercise training on the longissimus muscle types was investigated in rats. There were four groups of rats, each exposed to exercise of equal intensity, but of varying duration. A segment of the longissimus muscle between the 2nd and 4th lumbar vertebrae was used for muscle fiber classification with histochemical ATPase classification. With exercise there were no significant changes in the percentage of the muscle comprised of type I muscle fibers. There was, however, significant increase in the percentage of type II A fibers. This paralleled a decrease in type II B fibers. The percentage of FG fiber types decreased significantly in all groups studied. There was a correlation between intensity and duration of exercise and the degree of decrease seen in the latter fiber types. An increase in oxidative muscle capabilities was evidence by the changes observed in the subtypes of class II fibers.     

Adaptation of Equine Locomotor Muscle Fiber Types to Endurance and Intensive High Speed Training

Skeletal muscle constitutes the largest organ system in the mammalian body and is essential for movement and force generation. Muscle tissue has the unique ability to adapt and remodel with regular exercise. Adaptation of equine contractile apparatus to exercise training with a different character occurs at the structural to the cellular and molecular levels and depends on age, breed, and sex. In Andalusian and Arabian horses from 3 months to 24 years of age, it was found that the mean cross-sectional area occupied by IIA and IIX fibers was greater in stallions than in mares. In muscle of Dutch Warmblood foals from birth to 1 year of age, a significant number of fibers coexpress either developmental and type IIa or cardiac–alpha and type I myosin heavy chain (MyHC) isoforms. Endurance training results in increased mitochondrial density, capillary supply, changes in key metabolic enzymes, and increased maximal oxygen uptake and promotes a transition from type II to type I muscle fiber. In horses, prolonged aerobic exercise training has been shown to induce a further decline in the percentage of type IIx MyHC isoform expression and an increase of type I and IIa MyHC isoform expression. Short-duration, high-intensity exercise training stimulates type IIA and hybrid (IIA/IIX) fibers. Therefore, intensive high-speed trotting facilitates muscle fiber hypertrophy and increases the oxidative capacity of type IIX fibers.     

Association of a single nucleotide polymorphism in the 5' upstream region of the porcine myosin heavy chain 4 gene with meat quality traits in pigs

We identified a potential molecular marker associated with meat quality traits in the myosin heavy chain 4, MYH4 gene of Landrace pigs. Sequencing revealed a single nucleotide polymorphism (SNP; g.‐1398G>T) in the 5' upstream region of MYH4. It was significantly associated with the number of type IIa muscle fibers and water‐holding capacity based on filter‐paper fluid uptake. The GG genotype groups had a greater number of type IIa fibers and a larger area composed of type IIa fibers than the other genotype group (P = 0.004 and P = 0.061, respectively). Expression level of MYH4 gene in the genotype TT or GT was higher than in genotype of GG (P < 0.0001). The T allele may enhance expression level of MYH4 gene and then the portion of IIb type fiber in the muscle be increased by the T allelle. Therefore, we suggest that the g.‐1398G>T in the 5' upstream region of the porcine MYH4 may be used as a molecular marker for meat quality traits, although its functional effect is not defined yet.     

Histopathological observation of stress myopathy in M. longissimus in the pig and relationships with meat quality, fattening and slaughter traits

The characteristic myopathic features revealed by histological observations included strong proliferation of connective and fatty tissue, perivascular infiltrations and necrosis of muscle fibers with phagocytosis to the lesser extent. In the myopathic muscle, as well as in giant fibers, histochemical techniques showed a reduction in succinate dehydrogenase and lactate dehydrogenase activity in type beta R (slow-twitch, oxidative) and alpha R (fast-twitch, oxidative and glycolytic). Magnesium-activated adenosine triphosphatase reaction ranged from diffuse to negative in beta R, alpha R and alpha W (fast-twitch, glycolytic) fiber types. Diffuse reaction for acid phosphatase and total loss of glycogen content were observed. The micrographs of the myopathic muscle indicated enlarged mitochondria with atrophy or complete destruction of cristae. Many myofibrils were hypercontracted. Giant fibers possessed mitochondria enlarged to an even greater extent and many of the myofibrils had loss of continuity, were narrow, depleted and were also hypercontracted. Significant differences between myopathic and normal groups were found in number of beta R fibers (lower in the myopathic group), number of alpha R fibers and percent of alpha R and alpha W fibers (higher in the myopathic group). Differences (P less than .01) existed between meat pH1 value in the myopathic group (mean value of 5.95) and the normal group (mean value of 6.29). Meat from the myopathic group of pigs also had a lower (P less than .01) pH24 value and reduced water-holding capacity (P less than .01) relative to the meat of the normal pigs. The lack of difference of fattening and slaughter traits between the groups suggested that the White Zlotnicka pigs is of particular value because it is possible to improve the production traits without increasing the incidence of these syndromes within the breed. Negative correlations (P less than .05) between number of giant fibers and percent of alpha W fibers, and between percent of giant fibers and percent of alpha W fibers indicate that alpha W fibers can undergo degeneration and be transformed into giant fibers. Therefore, it it suggested that giant fibers should be treated as muscular, pathological results of past stresses and not as an additional type of normal muscle cells.     

Histochemical staining characteristics of normal horse skeletal muscle

The histologic and histochemical staining characteristics of the triceps brachii (long head), extensor carpi radialis, gluteus medius, vastus lateralis, biceps femoris, semimembranosus, semitendinosus, and extensor digitorum longus muscles of 8 Thoroughbreds, 2 Quarter Horses, 1 Arabian, 1 Paso Fino, and 1 Shetland Pony are described. Muscle fiber morphology, staining distribution and intensity, amount of IM connective tissue, number of IM blood vessels and IM nerves, calcium-activated adenosine triphosphatase activity (CaATPase), percentage of fibertype population, percentage of relative fibertype area, mean fiber diameter, nonspecific esterase activity, alkaline phosphatase activity, and acid phosphatase activity were evaluated, using 10 common histochemical and histologic stains. Two fiber types (I, II) and 3 subtypes (IIA, IIB, IIC) were observed, using CaATPase-, nicotinamide-adenine dinucleotide-tetrazolium reductase-, periodic acid-Schiff hematoxylin-, and nonspecific esterase-stained frozen serial muscle sections. Type I muscle fibers in general had low CaATPase activity, high oxidative capacity, low glycogen capacity, and low esterase activity. Type IIA muscle fibers had high CaATPase activity, intermediate oxidative capacity, high glycogen concentration, and high esterase activity. Type IIB fibers had high CaATPase activity, low oxidative capacity, high glycogen concentration, and a high esterase activity. Type IIC muscle fibers had high CaATPase activity, high oxidative capacity, variable glycogen concentration, and high esterase activity. Type II (IIA and IIB) muscle fibers predominated in the muscles. The percentage of muscle fiber population, mean minimal muscle fiber diameter, and percentage of relative muscle fiber area were determined for each sampled muscle. Type IIA and IIB muscle fibers predominated in the percentage of muscle fiber population and percentage of relative muscle fiber area. Type IIB muscle fibers had the greatest minimal fiber diameter, type IIA muscle fibers had intermediate minimal fiber diameter, and type I muscle fibers had the smallest minimal fiber diameter. The percentage of relative muscle fiber area was less variable (P less than or equal to 0.05) than the percentage of muscle fiber population. Mean muscle fiber diameter did not significantly differ between breeds. Alkaline and acid phosphatase activities were at low levels in all muscles biopsied and were limited to the IM connective tissue fibrocytes, macrophages, and capillaries.(ABSTRACT TRUNCATED AT 400 WORDS)     

Bovine sire selection based on maintenance energy affects muscle fiber type and meat color of F1 progeny

A total of 42 F(1) Red Angus progeny from sires divergent in maintenance energy (ME(M)) EPD were analyzed to determine whether selecting for sire ME(M) would alter end-product meat quality. Data from animals were grouped based on the divergence of the ME(M) EPD of their sire from the Red Angus Association-reported breed average and defined as either high or low, the assumption being that high-ME(M) cattle are less efficient because their maintenance requirements represent a larger proportion of their dietary intake. Steer progeny (n = 7) from the high group produced bottom round steaks with a greater a* (redness) color value (P = 0.02) after 5 d in a simulated retail display when compared with bottom round steaks from the low group (n = 18). Bottom round steaks from the high group had a greater b* (yellowness) color value at d 1 (P = 0.03) and d 5 (P = 0.01) of retail display. Samples from the biceps femoris were taken at 12 mo (from both steers and heifers) and 15 mo (from steers only) of age for fiber type proportion analysis. At 12 mo of age, steers from the low group had more type I fibers (P = 0.02), whereas steers from the high group had more type IIb fibers (P = 0.01). Furthermore, samples from steers in the low group at 15 mo had more type I fibers (P = 0.02), and steers from the high group maintained more type IIb fibers (P = 0.02). No changes in fiber type proportions were observed between the high- and low-ME(M) EPD heifers (n = 17). Relative mRNA abundance of genes involved in the synthesis, storage, and breakdown of glycogen were analyzed as a variable important for meat quality, but no statistical differences were observed. At 12 mo age, glycogenin (glyc) was negatively correlated with the proportion of type IIa fibers (r = -0.32 and P = 0.12) as well as with the proportion of type IIb fibers (r = -0.42 and P = 0.03) in the biceps femoris of the steers. In samples taken from the biceps femoris at 15 mo age, glyc was negatively correlated with the proportion of type IIa fibers (r = -0.42 and P = 0.03) in the steers. This indicates that relative mRNA expression of glyc may serve as a marker of muscle glycogen storage capacity in steers. Thus, selection for efficient Red Angus beef cattle based on sire ME(M) EPD does not adversely affect meat quality in F(1) progeny, based on the variables assessed in this study. Furthermore, selection for progeny from low-ME(M) EPD sires may improve fresh meat quality within Red Angus beef cattle.     

Correlation between histochemically assessed fiber type distribution and isomyosin and myosin heavy chain content in porcine skeletal muscles.

Highly sensitive enzyme assays developed to differentiate skeletal muscle fibers allow the recognition of three main fiber types: slow-twitch oxidative (SO), fast-twitch oxidative glycolytic (FOG), and fast-twitch glycolytic (FG). Myosin, the predominant contractile protein in mammalian skeletal muscle, can be separated based on the electrophoretic mobility under nondissociating conditions into SM2, SM1, IM, FM3, and FM2 isoforms, or under dissociating conditions into myosin heavy chain (MHC) I, IIb, IIx/d, and IIa. The purpose of the present study was to determine whether the histochemical method of differentiation of fiber types is consistent with the electrophoretically identified isomyosin and MHC isoforms. These comparisons were made using serratus ventralis (SV), gluteus medius (GM), and longissimus muscles (LM) from 13 pigs. Two calculation methods for the histochemical assessed fiber type distribution were adopted. The first method incorporated the number of fibers counted for each fiber type and calculated a percentage of the total fiber number (fiber number percentage: FNP). The second method expressed the cross-sectional area of each fiber type as a percentage of the total fiber area measured per muscle (fiber area percentage: FAP). Independent of the calculation methods, correlation analyses revealed in all muscles a strong relation between SO fibers, the slow isomyosin (SM1 and SM2), and MHCI, as well as between the FG fibers, the fast isomyosin (FM3 and FM2), and MHCIIx/b content (P<.05). There were no correlations between FOG fiber population assessed by histochemical analysis and intermediate isoform (IM) or MHCIIa content. The present results did not provide conclusive evidence as to which of the calculation methods (FNP or FAP) was more closely related to myosin composition of skeletal muscles. Despite some incompatibility between the methods, the present study shows that histochemical as well as electrophoretic analyses yielded important information about the composition of porcine skeletal muscle. The combination of the two methods may be essential to accurately characterize porcine skeletal muscles.     

Fiber type composition of rostral and caudal portions of the digastric muscle in the dog

The distribution of muscle fiber types in rostral and caudal portions of the musculus digastricus (digastric muscle) was studied in 6 dogs. Staining procedures which stain specifically for type IIM fibers, a fiber type found in other muscles supplied by the trigeminal nerve, were used. Rostral and caudal portions of the muscle were compared because the rostral portion is innervated by the trigeminal nerve, and the caudal portion is innervated by the facial nerve. The musculus triceps brachii (triceps muscle), which contains fiber types I and IIA, and the musculus masseter (masseter muscle), which contains type IIM, were used as controls. Mean fiber diameters were calculated for each of the muscles. Both portions of the digastric muscle exhibited the same histochemical behavior, possessing types I and IIA myofibers. Neither portion contained type IIM fibers. Type I fibers in the masseter muscle were histochemically different from type I fibers in the other muscles studied. Type II fibers predominated in all 3 muscles, but there were significantly (P less than 0.001) more type I fibers in the triceps muscle than in either portion of the digastric muscle or in the masseter muscle. Type II fibers were significantly larger than type I fibers in the caudal digastric (P less than 0.01) and masseter (P less than 0.05) muscles. There was no difference in the size of type I or type II fibers between any of the muscles studied (P greater than 0.20).     

Glut4突变调控脂肪重分布及肌纤维重塑的机制研究

旨在探讨Glut4基因突变后骨骼肌能量代谢及肌纤维转化的分子机制。本研究利用CRISPR/Cas9技术构建Glut4^Q177L突变鼠。选取22周龄健康的野生雄鼠和Glut4^Q177L突变雄鼠,即试验分为两组,每组3只,3个重复,进行表型评价和糖耐量、胰岛素耐量试验;荧光定量检测两组小鼠腓肠肌葡萄糖转运蛋白、脂代谢、肌纤维类型相关调控基因的表达差异;Western blot测定腓肠肌AMPK及其磷酸化蛋白含量。结果表明,突变鼠皮下脂肪和附睾脂重量显著低于对照组（P<0.05）;突变鼠糖耐量曲线下面积极显著增加（P<0.01）,表明其糖耐量受损;突变鼠血清中甘油三酯的含量显著下降（P<0.05）。相较于野生型小鼠,Glut4^Q177L突变鼠腓肠肌中Glut4、Glut1和Glut12等多个葡萄糖转运蛋白表达量显著升高（P<0.05）;葡萄糖摄取受限导致调控能量代谢关键基因AMPK在mRNA、蛋白和磷酸化修饰水平均显著升高（P<0.05）,同时促进与脂肪酸摄取与合成代谢相关的CD36和ATGL等基因表达上调（P<0.05）;慢速氧化型肌纤维（I型）和快速氧化型肌纤维（IIa型）相关基因表达极显著升高（P<0.01）,而快速酵解型纤维（IIb型）相关基因表达显著降低（P<0.05）。本研究结果显示,Glut4葡萄糖结合关键位点突变降低了骨骼肌和脂肪葡萄糖摄取能力,通过上调其它转运蛋白增加葡萄糖摄取;激活AMPK信号通路及分泌肌细胞因子调控脂肪分解以满足能量需求;促进线粒体丰富的氧化型肌纤维生成以提高能量利用效率。Glut4突变不仅可以为骨骼肌胰岛素抵抗提供有效的动物模型,还可以为家畜生物育种提供基因编辑参考位点。     

The effect of testosterone on gastrocnemius muscle fibres in growing and adult male and female rats: a histochemical,morphometric and ultrastructural study

In this study, the effect of testosterone on gastrocnemius muscle fibres in growing and adult rats (male and female) was examined using histochemical, morphometric and ultrastructural techniques. After physiological saline (PS), olive oil (OvO) or olive oil + testosterone (OvOT) injections on 72 rats (growing and mature, 36 male and 36 female), the sample tissues of fibre types of the gastrocnemius muscle taken were examined by histochemical [alkaline adenosine triphosphatase (alk-ATPase), acid ATPase (ac-ATPase)], morphometric and ultrastructural techniques. In PS-injected control groups, the gastrocnemius muscle of both sexes contained all the fibre types studied [slow-oxidative muscle fibres (type I), fast-oxidative glycolytic muscle fibres (type IIA) and fast-glycolytic muscle fibres (type IIB)]. The type I fibres had the smallest diameter, type IIA had a medium diameter and type IIB fibres had the largest diameter. In OvO-injected groups, it was observed that the OvO had little effect on the gastrocnemius muscles of either sex, although there was significant enlargement of type IIB fibres. After the injection of OvOT, hypertrophy of muscle fibres was determined by morphometric study. The biggest increase in diameter was on type I fibres. In addition, degenerations on some mitochondria, accumulation of lipid droplets on type I and type II fibres, an increase in glycogen particles, bifurcation of myofibrils, an increase in the number and diameter of units resembling T tubules and an increase in ribosomal content were also observed in the same group by transmission electron microscope. Consequently, it was determined that testosterone can induce protein synthesis in gastrocnemius muscle fibres, and induces changes in shape and size, and also can change the appearance and the number of fibres.     

Muscle fibre distribution in forelimb,hindlimb and trunk muscles in three bat species: The little Japanese horseshoe,greater horseshoe and Egyptian fruit

This study characterised muscle fibres in trunk, forelimb and hindlimb muscles of three bat species: little Japanese horseshoe (Rhinolophus cornutus), greater horseshoe (Rhinolophus ferrumequinum) and Egyptian fruit (Rousettus aegyptiacus). Twenty-seven muscles from trunk, forelimb and hindlimb were dissected, weighed and analysed by immunohistochemistry and sodium didecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and determined their cross-sectional areas (CSA). Results showed that Type IIa and Type IIa/x made the highest proportion of total muscle mass. Moderate proportion was formed by Type IIb. Type I and IIx appeared at very low levels in all bats. Type IIb was the only fibre type detected in patagial muscles in wing membrane of greater horseshoe while other fibre types were not observed. Type I muscle fibres were very few and appeared infrequently in fifteen muscles of Egyptian fruit and in only one muscle in each, greater horseshoe and little Japanese horseshoe. Type IIx was also detected in three muscles in greater horseshoe and only one muscle in Egyptian fruit but none in little Japanese horseshoe. The highest average CSA μm² was detected in Type IIb and values were 734.2μm² for LHB; 1537.9μm² for GHB and 1,720.9μm² for EFB. Lowest and undetermined values were observed for Type I and IIx. These data demonstrate that Type IIa, IIa/x and IIb form significant proportion of adult bat muscle mass and Type IIb is the largest fibre type. The distribution pattern is suggestive of specialised functions of the fibres in relation to orientation and speed of bats during flight.     

Semitendinosis muscle development in several strains of fetal and perinatal pigs

At 110 d of gestation, fetuses were removed from Ossabaw, Yorkshire and crossbred sows and from sows selected for high (obese) or for low (lean) backfat thickness. Ossabaw and obese fetuses were smaller than lean, Yorkshire and crossbred fetuses (595 +/- 32 and 863 +/- 44 g vs 1,030 +/- 77, 1,380 +/- 1,144 +/- 80 g; means +/- SE), respectively. Minimum fiber diameters in the semitendinosus muscle were larger in obese, lean and Ossabaw fetuses than in Yorkshire and crossbred fetuses (12.9 +/- .3, 12.5 +/- .2 and 11.8 +/- .2 micron vs 10.2 +/- .2 and 11.1 +/- .8 micron), respectively. Histochemical analysis for NADH-tetrazoleum reductase (NADH-TR) and esterase activities indicated no fiber type differentiation and no strain differences. Fiber type differentiation was obvious with acid ATPase histochemistry in muscles from all fetuses. The white portion of the semitendinosus from Ossabaw, obese and lean fetuses had many fibers that contained no histochemically detectable lipid (oil red O staining). The unstained fibers (oil red O) were always the most peripherally located fibers in a fasciculi. In some instances, 50% of the fibers in a fasciculi were not stained for lipid. All the fibers in the red portion of muscle from Ossabaw, obese and lean fetuses contained lipid. All the fibers in the red and white portions of muscles from crossbred and Yorkshire fetuses contained lipid. Muscles from young (1 to 2 d old) Ossabaw, Yorkshire and crossbred pigs were also histochemically analyzed. Analysis for NADH-TR, esterase and alkaline phosphatase (capillary staining) activities indicated no fiber type differentiation and no strain differences. As in the fetuses, the white portion of muscle from Ossabaw pigs had many fibers with no lipid (oil red O). Lipid was present in all fibers in the deep portions of muscle from Ossabaws and in all fibers in both portions of muscle from crossbred and Yorkshire pigs. These results indicate that when lipid staining is used as the criterion, fiber type differentiation is evident in muscle from fetuses and young pigs from strains not genetically selected for muscling (Ossabaw, obese and lean strains). Furthermore, fiber type differentiation is not evident in muscle from strains of pigs genetically selected for greater muscling (crossbred and Yorkshires).     

Meta-analysis of the effect of animal maturity on muscle characteristics in different muscles, breeds, and sexes of cattle

The effect of animal maturity on fiber cross-sectional area, percentage of fiber types, activities of isocitrate dehydrogenase (ICDH) and lactate dehydrogenase (LDH), total and insoluble collagen and lipid concentration was investigated in the longis-simus thoracis (LT), semitendinosus (ST), and triceps brachii (TB) muscles. The analysis considered 2,642 muscle samples from bulls, steers, and cows of Aubrac, Charolais, Limousin, Montbéliard, and Salers breeds. For the bulls, the fiber cross-sectional area, percentage of slow oxidative fibers, and ICDH activity showed a quadratic relationship (P < 0.05), and the percentage of fast oxidative-glycolytic and fast glycolytic fibers and LDH activity showed a cubic relationship (P < 0.05) with increased maturity. A linear relationship was observed for the collagen and lipid muscle characteristics. The response equation coefficients for different muscles indicate that development of muscle characteristics is different for each muscle. Compared with the other muscles, ST muscle had a greater fiber cross-sectional area, proportion of fast glycolytic fibers, LDH activity, and collagen content. The LT muscle had a greater proportion of slow-oxidative fibers and lipid (P < 0.05). Within the ST muscle, all characteristics except lipid concentration showed different development between the breeds. Steers showed greater changes in muscle fiber cross-sectional area, percentage of fast oxidative-glycolytic and fast glycolytic fibers, and total lipid in the muscle with increasing maturity compared with bulls. The mean fiber cross-sectional area and percentage of fast glycolytic fibers was greater and the mean lipid concentration was less in bulls compared with steers (P < 0.05). Data for cows were from more mature animals. Muscle characteristics in cows did not show large changes with increasing degree of maturity. Muscle type accounts for a greater proportion of the variation in the muscle characteristics than breed and sex of the animal.     

Ultrastructural alterations of the myocardium of rats fed rapeseed oils.

Ultrastructural studies were conducted on the myocardium of rats fed corn oil, Tower RSO (0 . 88 per cent EA), 1788 RSO (3 . 6 per cent EA) and Target RSO (38 . 9 per cent EA) supplemented diets for 18 weeks. Cardiac myocytes of Tower RSO-fed rats showed some loosening of myofibrils and a slight increase in the number of mitochondria, few of which had lost their cristae. Large intravascular lipid droplets were observed in the myocardium of rats fed the 1788 RSO diet, as well as some small lipid droplets which were seen in close association with mitochondria. There was an apparent increase in the number of mitochondria of both normal and giant size. Many of the mitochondria exhibited distortion of shape and degeneration of cristae. The matrix of megamitochondria contained vesicles and electron-dense floccular inclusions and at times electron-lucent lipid-like material. The degenerative changes of mitochondria were most pronounced in the Target RSO group, where some megamitochondria showed a complete loss of cristae and a replacement of matrix with lipid-like material. These observations suggest that both intravascular lipid globules and the mitochondrial alterations are possible contributory factors involved in the development of cardiac lesions in RSO-fed rats.