Protective Effects of Voluntary Exercise on Hepatic Fat Accumulation Induced by Dietary Restriction in Zucker Fatty Rats.

Weight control based on dietary restriction (DR) alone can cause lipid metabolic failure and progression to fatty liver. This study aimed to investigate the effect of exercise on preventing DR-induced hepatic fat accumulation in Zucker fatty (ZF) rats by focusing on the relationship between adipose tissue lipolysis and hepatic fat uptake. Six-week-old male ZF rats were randomly assigned to obese, DR, or DR with exercise (DR + Ex) groups. The DR and DR + Ex groups were fed a restricted diet, with the latter also undergoing voluntary exercise. After 6 weeks, hepatic fat accumulation was observed in the DR group, whereas intrahepatic fat was markedly reduced in the DR + Ex group. Compared with the obese (Ob) group, the DR group exhibited 2.09-fold expression of hepatic fatty acid translocase (FAT)/CD36 proteins (p < 0.01) and 0.14-fold expression of hepatic fatty acid-binding protein (FABP)1 (p < 0.01). There were no significant differences between the DR + Ex group and the Ob group. FAT/CD36 and hepatic triglyceride (TG) expression levels were strongly positively correlated (r = 0.81, p < 0.001), whereas there was a strong negative correlation between FABP1 and hepatic TG expression levels (r = -0.65, p < 0.001). Our results suggest that hepatic fat accumulation induced by DR in ZF rats might be prevented through exercise-induced modifications in FAT/CD36 and FABP1 expression.

Characterization of fat metabolism in the fatty liver caused by a high-fat, low-carbohydrate diet: A study under equal energy conditions.

The pathology of fatty liver due to increased percentage of calories derived from fat without increased overall caloric intake is largely unclear. In this study, we aimed to characterize fat metabolism in rats with fatty liver resulting from consumption of a high-fat, low-carbohydrate (HFLC) diet without increased caloric intake. Four-week-old male Sprague-Dawley rats were randomly assigned to the control (Con) and HFLC groups, and rats were fed the corresponding diets ad libitum. Significant decreases in food intake per gram body weight were observed in the HFLC group compared with that in the Con group. Thus, there were no significant differences in body weights or caloric intake per gram body weight between the two groups. Marked progressive fat accumulation was observed in the livers of rats in the HFLC group, accompanied by suppression of de novo lipogenesis (DNL)-related proteins in the liver and increased leptin concentrations in the blood. In addition, electron microscopic observations revealed that many lipid droplets had accumulated within the hepatocytes, and mitochondrial numbers were reduced in the hepatocytes of rats in the HFLC group. Our findings confirmed that consumption of the HFLC diet induced fatty liver, even without increased caloric intake. Furthermore, DNL was not likely to be a crucial factor inducing fatty liver with standard energy intake. Instead, ultrastructural abnormalities found in mitochondria, which may cause a decline in ß-oxidation, could contribute to the development of fatty liver.

Effects of habitual exercise and dietary restriction on intrahepatic and periepididymal fat accumulation in Zucker fatty rats.

BACKGROUND: Habitual exercise and dietary restriction are commonly recommended to prevent or ameliorate obesity and lifestyle-related diseases, including fatty liver. This study investigated the effects of habitual exercise and dietary restriction on hepatic triglyceride (TG) levels, serum leptin levels, and histological adipocyte size in periepididymal adipose tissue from Zucker fatty (ZF) rats. METHODS: Six-week-old male ZF rats were randomly assigned to one of three groups: sedentary (Sed), sedentary and dietary restriction (Sed + DR), and training and dietary restriction (Tr + DR). Male Zucker lean (L) rats were used as control animals. All rats had access to water and the allowed quantity of food ad libitum. The rats in the Sed + DR and Tr + DR groups were fed a 30% restricted diet, while those in the Tr + DR group exercised voluntarily on a wheel ergometer. After 12 weeks, the rats were sacrificed for a histological examination of their liver and periepididymal adipose tissue. Hepatic and serum TG, serum total cholesterol, glutamic-oxaloacetic transaminase, glutamic-pyruvic transaminase, free fatty acid, and leptin levels were also measured. RESULTS: The hepatic TG levels were significantly higher in the Sed + DR group than in the L (P < 0.001) and Sed (P < 0.05) groups. By contrast, the hepatic TG levels in the Tr + DR group were significantly lower than those in the Sed (P < 0.05) and Sed + DR (P < 0.001) groups, but not significantly different from the L group values. The periepididymal adipocytes were significantly larger in the Sed, Sed + DR, and Tr + DR groups than in the L group (P < 0.001) and were significantly smaller in the Tr + DR group compared to the Sed and Sed + DR groups (P < 0.001). CONCLUSIONS: Our results suggest a relationship between lipid metabolism and the size of adipose cells in ZF rats. Exercising plays an important role in decreasing hepatic TG levels, serum leptin levels, and the size of adipose cells.

Filamentous structure of the external surface of abalone eggs revealed by quick-freeze deep-etch technique.

The external surface of abalone eggs was examined by thin section and quick-freeze, deep-etch electron microscopy. In thin sections, networks of fine filaments were found interconnecting the adjacent microvilli on the surface of unfertilized eggs. Quick-freeze, deep-etch electron microscopy revealed the three-dimensional structure of these networks of filaments on the external surface of the egg. Mainly two networks of filaments were identified; one was composed of thicker (14-19 nm) filaments interconnecting with the neighboring microvilli nearly horizontally, and the other was composed of thinner (8-14 nm) branched filaments closely surrounding the microvilli surface as well as highly interconnecting neighboring microvilli in a polygonal pattern. The overall structure of the filamentous network on the egg surface showed no distinct alteration after fertilization. These networks of filaments observed on the egg surface may play a key role in sperm-egg interaction.

Organization of Actin Filaments in the Axial Rod of Abalone Sperm Revealed by Quick Freeze Technique: (actin filaments/abalone sperm/quick-freeze deep-etch/freeze-substitution/cytoskeleton).

An axial rod in abalone (Haliotis discus) sperm is a structure composed of a bundle of actin filaments, which elongates anteriorly to form the acrosomal process during the acrosome reaction. The ultrastructure of the actin filament bundle constituting the axial rod was examined using quick freeze technique followed by either freeze-substitution or deep-etch electron microscopy. Thin sections of quick freeze and freeze-substituted sperm revealed that the actin filaments in the axial rod are hexagonally packed in a paracrystalline array through its almost entire length with an average center-to-center spacing of 12 nm. Periodic transverse bands were also observed across the actin filament bundle, which may reflect the cross-bridges interconnecting the adjacent filaments. Quick-freeze deep-etch analysis provided the three-dimensional view of the axial rod. Actin filaments exhibiting 5.5-6 nm spaced striations were observed to run in parallel with each other inside the axial rod. The existence of cross-bridging structures was also displayed between adjacent filaments. These results suggest that the actin filaments in the axial rod are probably held together by regularly spaced cross-bridges to form a well ordered hexagonally packed bundle, and also cross-linked by fibrous structure to the lateral inner acrosomal membrane which closely surrounds the anterior half of the actin filament bundle.

Coiled Filamentous Structure in the Sperm Acrosome of Gastropoda, Sulculus aquatilis and Turbo cornutus: (acrosome reaction/coiled filaments/gastropoda/cytoskeleton/fertilization).

A helically coiled filamentous structure, termed the truncated cone originally identified in Haliotis discus, was demonstrated to exist in the apex of the acrosome subjacent to the outer acrosomal membrane of the sperm in two species of Gastropoda, Sulculus aquatilis and Turbo cornutus. Thin section and whole mount preparations revealed that in intact sperm this filamentous structure consisted of about 12 helically arranged filaments with a diameter of 10-12 nm which were tightly packed in a truncated shape. This truncated cone elongated anteriorly from the acrosomal opening and transformed into a cylinder which closely surrounds the acrosomal process during the acrosome reaction. In S. aquatilis and T. cornutus, the truncated cone elongated more than 3 and 1.5 times as long as the original height, respectively. The elongated truncated cone was characterized by striations with increase in its periodic spacing and inclinations in thin sections. The truncated cone in both the species was fundamentally analogous to that of Haliotis discus, further suggesting that the truncated cone plays a role in fertilization as a common cytoskeletal structure among the species of Gastropoda.

Distribution of Intramembrane Particles and Its Changes during the Acrosome Reaction in Spermatozoa of the Japanese Abalone: (abalone sperm/acrosome reaction/membrane particle/freeze-fracture).

The distribution of intramembrane particles in the plasma and acrosomal membranes of sperm of the Japanese abalone, Haliotis discus, and its changes during the acrosome reaction were studied by the freeze-fracture replica technique. The P face of the plasma membrane covering the acrosome has sparse membrane particles except in the apical region, which includes the trigger and 'truncated cone' regions. Large particles with an average diameter of 10 nm are located in this apical region. The E face of the plasma membrane has only a few particles. On the outer acrosomal membrane, many particles are randomly distributed throughout the P face, but only a small number of particles are found on the E face. Numerous particles on the P face of the inner acrosomal membrane show a regular arrangement as a dense lattice or with a concentric circular pattern. The initial change in the acrosome reaction is clearance of membrane particles from both the P and E faces of the plasma and outer acrosomal membranes around the apical region, where fusion of the two membranes occurs. As the acrosomal process elongates, the dense arrangement of particles on the inner acrosomal membrane changes via a loose lattice arrangement to a patchy distribution with particle-free areas. Then the arrangement is further disorganized becoming a sparse, random distribution.

Ultrastructural Changes of the "Truncated Cone" during the Acrosome Reaction in Japanese Abalone Spermatozoa: (abalone/acrosome reaction/truncated cone/ultrastructure).

An electron-dense structure termed the "truncated cone" covers the apical surface of the acrosomal contents except for the trigger region in Haliotis discus spermatozoa. The truncated cone, having a slant height of 0.3 µm and diameters of circular top and base of 0.3 and 0.6 µm, shows striations with a periodicity of 6.6 to 8.0 nm. During the acrosome reaction, the truncated cone elongates simultaneously with the protrusion of the acrosomal process through the truncated cone. As the growth of the acrosomal process further proceeds, the truncated cone transforms into a cylindrical shape and eventually reaches 1 µm in length and 0.2 µm in diameter. The elongated truncated cone is characterized by regularlly helical striations with a periodicity of 19 to 21 nm with an inclination of 40° to 46°. These results may suggest that the truncated cone is composed of coiled filaments, which coil up further during the acrosome reaction causing the truncated cone to slenderize and elongate. The elongation is also achieved by stretching of the coil. In H. discus hannai Ino, structural changes in the truncated cone show close homology to those in H. discus. No such morphologically unique organelle has been found in other species thus far.

Fine Structural Changes in the Acrosome Reaction of the Japanese Abalone, Haliotis disus: (abalone/sperm/acrosome reaction/ultrastructure).

The spermatozoon of the Japanese abalone, Haliotis discus, and its structural changes during the acrosome reaction were observed by electron microscopy. The spermatozoon has a huge acrosome in the shape of a hanging bell or a forefinger with a deep fossa at the posterior end being filled with a bundle of microfilaments. The membranes of the acrosomal apex, the so-called trigger region, are structurally discernible from those of other acrosomal regions. Following the trigger region, a unique structure under the acrosomal membrane covers the surface of the acrosomal content in the form of a truncated cone. The acrosome reaction occurs in the jelly layer very close to the egg envelope. First, the membranes at the apex of the acrosome are vesiculated, followed by the formation of a narrow gap between the outer acrosomal membrane and the acrosomal content. Next, the bundle of micro-filaments elongates, running through the center of the acrosome, reaching the trigger region and protruding out of the acrosomal top. Then release of the acrosomal content occurs in two steps, disclosing the "membrane undercoating structure" that comprises globular particles with a fuzzy material connecting them. This resembles the undercoat network found in erythrocytes.