The fine structure of the liver cells in the bat (Myotis myotis) during hibernation,arousal and forced feeding

Summary The ultrastructure of liver cells was studied in three groups of bats (Myotis myotis) captured in March. 1. Hibernating animals: The liver cells contain much glycogen and large mitochondria, poorly developed ER, relatively well developed Golgi apparatus, various amounts of lysosomes. Most of the bile canaliculi are closed. 2. Non-hibernating, starving animals: 24 hours after arousal the glycogen and lipid droplets disappear from the liver cells, the mitochondria swell slightly. There is no essential alteration in the ER. In the liver cells of the starving animals numerous autophagic vacuoles appear including disintegrated cytoplasmic components. 3. Non-hibernating force-fed animals: They were given food rich in fat and protein, containing sugar and a drug increasing bile secretion. Within an hour, glycogen and lipid reappears in the liver cells, autophagic vacuoles disappear, mitochondria become larger. After feeding, the Golgi apparatus hypertrophies considerably, many bile canaliculi open, the number of the peribiliary lysosomes decreases.This paper is dedicated to Professor W. Bargmann on the occassion of his 60th birthday.     

Ultrastructural study of liver cells from rooks living in ecologically unfavorable areas

The ultrastructure of liver cells was studied in rooks (Corvus frugilegus) living in radioactive and chemical contamination areas. The ultrastructure of liver cells from rook as well as jackdaw (Corvus monedula) and hooded crow (Corvus cornix) (Corvidae family) from a conventionally clean area was studied as control. Control hepatocytes proved to contain a great number of mitochondria, many of which were swollen and had clear matrix and disorganized cristae. The cristae nearly lacked glycogen and had abundant lipid droplets, which often tightly contacted mitochondria. The cytoplasm of hepatocytes in birds from both ecologically unfavorable areas had numerous mitochondria with the same ultrastructure. In contrast to control, the hepatocyte cytoplasm: (1) contained a lot of glycogen; (2) there were many lipid droplets, which directly contacted glycogen granules; and (3) had more abundant peroxisomes. In addition to normal erythrocytes, the sinusoids contained erythrocytes with mitochondria, vesicles, and lipid droplets in their cytoplasm. Analysis of many micrographs of lipid droplets contacting glycogen granules, mitochondria, peroxisomes, and cisterns of smooth endoplasmic reticulum allowed us to propose that glycogen is synthesized via gluconeogenesis from glycerol and products of fatty acid oxidation in the liver cell cytoplasm of rooks from ecologically unfavorable areas as distinct from control.     

Closer association of mitochondria with lipid droplets in hepatocytes and activation of Kupffer cells in resveratrol-treated senescence-accelerated mice

Resveratrol has been extensively investigated because of its beneficial effects in delaying age-related diseases, thus extending the lifespan, possibly by mimicking calorie restriction. For this study, cell biological techniques were used to examine how resveratrol influenced hepatocytes in a senescence-accelerated mouse P10 (SAMP10), treated from 35 to 55 weeks of age, with special emphasis on the relationship between mitochondria and lipid droplets. Survival ratio, body weight and food intake of SAMP10 did not differ significantly between the control and resveratrol-treated groups. Compared with the control, the treated livers were altered significantly, as follows. Lipid droplets were reduced and mitochondria were increased in number in hepatocytes. Phosphorylation of acetyl-CoA carboxylase and the expression of both the mitochondrial ATP synthase β subunit and Mn superoxide dismutase (SOD2) were increased. Mitochondria, expressing more SOD2, were more tightly associated with lipid droplets, suggesting the enhancement of lipolysis through the activation of mitochondrial functions. Cathepsin D expression was less in hepatocytes but enhanced in Kupffer cells, which were increased in number and size with more numerous lysosome-related profiles. Together, resveratrol may activate mitochondria resulting in consuming lipids, and may also activate Kupffer cells by which a beneficial milieu for hepatocytes may be created. Both might be related to improvement in the functioning of the liver, which is the organ that is central to metabolic regulation.     

The ultrastructure of the midgut glands inLigia italica (Isopoda) under different nutritional conditions

After a period of food deprivation,Ligia italica were refed for 2 days with different diets and their midgut glands were examined under the electron microscope with special reference to the large cells. The predominant features are the following: extended glycogen fields after sucrose-diet; numerous lipid droplets and peroxisome-like vesicles after lipid-diet (butter); swollen mitochondria and a great number of pinocytotic vesicles after protein diet (curds); electron dense vesicles and myelin bodies after the uptake ofEscherichia coli. In contrast to amphipods, the intertidal isopodL. italica is not able to digest cellulose, as the cell ultrastructure exhibits all features of starved animals, as well as that following feeding with lignin.Supported by Deutsche Forschungsgemeinschaft (Sto 75/4–9)     

Biochemical and Ultrastructural Changes in Tetrahymena pyriformis During Starvation*

SYNOPSIS. Certain of the ultrastructural and biochemical changes occurring during the first 25 hr of starvation in Tetrahymena pyriformis were studied. Ultrastructurally, numerous profiles of degenerating mitochondria were seen in the early stages of starvation. The presence of oxidizable substrate such as glucose and acetate did not prevent this degeneration. Numerous large nucleoli were formed, many of which seemed to be passing into the cytoplasm as forming autophagic vacuoles. There was a transient increase in Oil Red O-positive bodies, presumably lipid (triglycerides). The extent and duration of this increase were pronounced in the presence of acetate. The lipid droplets appeared to arise within the cisternae of the endoplasmic reticulum. Lipid reserves were apparently utilized prior to carbohydrates, as the disappearance of lipid droplets preceded glycogen utilization, both in the presence of acetate and in the absence of exogenous substrate. A considerable loss of cellular protein also occurred. In cells from inorganic medium supplemented with glucose, glycogen occupied much of the cell, leaving only islands of cell organelles. Acid phosphatase was localized, ultrastructurally, mainly in autophagic vacuoles which contained mitochondria and other cell organelles, and in association with small, double-membraned structures which seemed to be sequestering small areas of cytoplasm. Such sequestered areas also appeared within larger autophagic vacuoles. Residual bodies containing concentric whorls of myelin-like membranes surrounding a more solid core accumulated during starvation. Acid phosphatase activity decreased in amount but not in specific activity. The specific activity of cathespin doubled or tripled, but there was little change in total enzyme.     

The hepatocyte ultrastructure of the Sevan trout (Salmo ischchan gegarkuni Kessel) in ontogeny]

Hepatocytes of Lake Sevan Salmo were examined at several stages in their life cycles which are different from the point of view of the manner of feeding. Salmo were reared at the fish farm, they were fed with the yolk of the chick eggs. It was revealed that hepatocytes of larva, which was sampled immediately after hatching (endogenous feeding) intensively synthesized the proteins; accumulated and secreted the bile product stored glycogen and lipids. The ultrastructure of larva hepatocytes changed on the 5th and 10th day after larva began to accept food (mixed feeding--endogenous and exogenous). Golgi complex became bigger, glycogen disappeared, lipid droplets became smaller (on the 5th day) and disappeared completely (on the 10th day). Morphological differentiation finishes during the fingerling period (exogenous feeding). Cisternae of granular endoplasmic reticulum (GER) and mitochondria are arranged around nucleus, near bile canaliculus and sinusoids. Big areas of glycogen lie between the organelles. Relative volumes of GER, mitochondria, glycogen increased, but the relative volume of Golgi complex diminished.     

Zonal expression of StARD1 and oxidative stress in alcoholic-related liver disease

Alcoholic-related liver disease (ALD) is one of the leading causes of chronic liver disease and morbidity. Unfortunately, the pathogenesis of ALD is still incompletely understood. StARD1 has emerged as a key player in other etiologies of chronic liver disease, and alcohol-induced liver injury exhibits zonal distribution. Here, we report that StARD1 is predominantly expressed in perivenous (PV) zone of liver sections from mice-fed chronic and acute-on-chronic ALD models compared to periportal (PP) area and is observed as early as 10 days of alcohol feeding. Ethanol and chemical hypoxia induced the expression of StARD1 in isolated primary mouse hepatocytes. The zonal-dependent expression of StARD1 resulted in the accumulation of cholesterol in mitochondria and increased lipid peroxidation in PV hepatocytes compared to PP hepatocytes, effects that were abrogated in PV hepatocytes upon hepatocyte-specific Stard1 KO mice. Transmission electron microscopy indicated differential glycogen and lipid droplets content between PP and PV areas, and alcohol feeding decreased glycogen content in both areas while increased lipid droplets content preferentially in PV zone. Moreover, transmission electron microscopy revealed that mitochondria from PV zone exhibited reduced length with respect to PP area, and alcohol feeding increased mitochondrial number, particularly, in PV zone. Extracellular flux analysis indicated lower maximal respiration and spared respiratory capacity in control PV hepatocytes that were reversed upon alcohol feeding. These findings reveal a differential morphology and functional activity of mitochondria between PP and PV hepatocytes following alcohol feeding and that StARD1 may play a key role in the zonal-dependent liver injury characteristic of ALD.     

Energy reserves during food deprivation and compensatory growth in juvenile roach: the importance of season and temperature

The effect of 21 days of starvation, followed by a period of compensatory growth during refeeding, was studied in juvenile roach Rutilus rutilus during winter and summer, at 4, 20 and 27° C acclimation temperature and at a constant photoperiod (12L : 12D). Although light conditions were the same during summer and winter experiments and fish were acclimated to the same temperatures, there were significant differences in a range of variables between summer and winter. Generally winter fish were better prepared to face starvation than summer fish, especially when acclimated at a realistic cold season water temperature of 4° C. In winter, the cold acclimated fish had a two to three‐fold larger relative liver size with an approximately double fractional lipid content, in comparison to summer animals at the same temperature. Their white muscle protein and glycogen concentration, but not their lipid content, were significantly higher. Season, independent of photoperiod or reproductive cycle, was therefore an important factor that determined the physiological status of the animal, and should generally be taken into account when fish are acclimated to different temperature regimes. There were no significant differences between seasons with respect to growth. Juvenile roach showed compensatory growth at all three acclimation temperatures with maximal rates of compensatory growth at 27° C. The replenishment of body energy stores, which were utilized during the starvation period, was responsible for the observed mass gain at 4° C. The contribution of the different energy resources (protein, glycogen and lipid) was dependent on acclimation temperature. In 20 and 27° C acclimated roach, the energetic needs during food deprivation were met by metabolizing white muscle energy stores. While the concentration of white muscle glycogen had decreased after the fasting period, the concentrations of white muscle lipid and protein remained more or less constant. The mobilization of protein and fat was revealed by the reduced size of the muscle after fasting, which was reflected in a decrease in condition factor. At 20° C, liver lipids and glycogen were mobilized, which caused a decrease both in the relative liver size and in the concentration of these substrates. Liver size was also decreased after fasting in the 4° C acclimated fish, but the substrate concentrations remained stable. This experimental group additionally utilized white muscle glycogen during food deprivation. Almost all measured variables were back at the control level within 7 days of refeeding.     

非酒精性脂肪肝病肝组织超微结构特点

目的:阐明非酒精性脂肪肝病(NAFLD)的超微结构特点。方法:收集我校和其他单位送检的3例单纯性非酒精性脂肪肝,16例NASH患者和4例NAFLD肝硬化患者的肝穿刺组织。用2.5%戊二醛、1%锇酸双固定、Epon 812包埋,超薄切片70nm,醋酸铀和柠檬酸铅染色后,JEM-2000EX透射电镜观察。结果:单纯性脂肪肝患者主要表现为大小不等的脂滴沉积、以小脂滴为主,可互相融合。NASH患者的肝细胞都可出现大量脂滴积聚,为大小脂滴混合型、内容物主要为中等电子密度、比较均一的甘油三酯,部分脂滴周围可见磷脂成分,NASH患者肝细胞内脂滴也互相融合。肝细胞线粒体的超微结构改变包括多形性线粒体、基质颗粒增多、线粒体增大和嵴的丧失是主要的电镜异常发现,线粒体内还可见副晶格样包涵体。部分NASH患者肝细胞内可见Mallory小体。NASH患者肝细胞周围可见淋巴细胞浸润。肝血窦Kupffer细胞增生不明显,NAFLD肝硬化患者Disse间隙和肝细胞间可见胶原纤维增生。结论:NAFLD具有较为明确的超微结构改变,电镜检查有助于诊断。     

Seasonal changes in hepatocyte ultrastructure correlated with the cyclic synthesis of secretory proteins in the winter flounder (Pleuronectes americanus)

Liver tissue was sampled from flounder (Pleuronectes americanus) throughout the year with the intention of documenting changes in the ultrastructure coincident with the production and secretion of antifreeze proteins. In the winter, hepatocytes are dedicated to the production of these proteins and, in the female, also reproductive proteins. In both sexes, liver cells in the summer contain abundant lipid and glycogen stores. In the female, there is a conspicuous hepatocyte transformation from a fat-filled cell in the summer to one with well-developed rough endoplasmic reticulum in the winter. Large amounts of rough endoplasmic reticulum (11.2 mg/gm) were recovered after subcellular fractionation of female wintertime liver. The increased appearance of secretory organelles and the high number of nucleolar profiles observed in winter animals is consistent with the elevated demand for protein secretion and synthesis in both sexes. The fractional volumes occupied by lipid droplets and mitochondria were different when comparisons were made between sex and season. Females contained a greater volume of lipid than did males, and summer animals contained more lipid than those in winter.     

Ultrastructure and lipid content of the liver of the zebrafish,Brachydanio rerio,related to vitellogenin synthesis

The female zebrafish is capable of producing mature eggs on the fifth day of each reproductive cycle. During this five-day period the ultrastructure of hepatocytes undergoes several changes. The number of nuclear pores increases rapidly during spawning, followed by a proliferation of RER within 24 h. Two days after spawning, glycogen has disappeared and the liver contains large amounts of lipids. The lipid droplets are closely surrounded by elongated mitochondria. Golgi complexes are abundant, secreting dense bodies. Four days after spawning the hepatocytes tend to regain their pre-spawning appearance. It is suggested that the changes in the hepatocytes, which coincide with special phases of ovarian activity, are related to vitellogenin synthesis. Steroids, especially estradiol-17beta, may trigger this process in the liver.     

Morphology of the green livers in upstream migrants of Petromyzon marinus L.

A morphological comparison was made of the green livers of male and female lampreys (Petromyzon marinus L.) collected during the upstream (prespawning) migration. Light and electron microscope histochemistry for iron, and both thin sections and freeze-fracture replicas in the electron microscope, revealed some sexual dimorphism in these livers. Ferric iron is much more abundant in the liver of females and is present in the cytoplasmic matrix, in dense bodies, and in vacuoles of hepatocytes. The numerous vacuoles of females may be the deposition site of biliverdin and other bile components that would account for the darker green coloration of the liver compared to males. Hepatocytes in females are also characterized by prominent rough endoplasmic reticulum and Golgi apparatus that reflect the involvement of the cells in vitellogenesis. The presence of numerous lipid droplets in the hepatocytes of males indicates that the liver is an important storage site for fat. The lipid droplets are associated with electron-dense deposits of unknown nature. Large gap junctions typify the parenchymal cells of both male and female livers. Perisinusoidal and sinusoidal cells are similar to those in the nonparenchymal region in other vertebrate livers, namely, endothelial and Kupffer cells, lipocytes (Ito), and some granulated cells. The relationship of lipocytes to fibrous tissue and fibrogenesis is discussed.     

Ultrastructural changes in the gastrodermal phagocytic cells of the planarian Dugesia gonocephala s.l. during food digestion (Plathelminthes)

Summary In the present report the functional morphology of the planarian gastrodermal phagocytic cells is examined in feeding animals. A functional interpretation of some of the morphological findings is given. The events in the fine-structure modifications of the phagocytic cells in the course of phagocytosis and intracellular digestion of food particles were followed through five post-feeding stages in the planarian Dugesia gonocephala. Light and electron microscopical observations demonstrate that there is preliminary intraluminal digestion of food particles; their phagocytosis takes place quickly.Beef hepatocytes that served as food are found engulfed at first in food vacuoles near the apical border of the phagocytic cells, and are clearly recognizable. The vacuoles increase in number to occupy most of the cytoplasm of these cells. Progressive breakdown and disappearance of phagocytosed hepatocytes occurs. In time the vacuoles move deeper into the cells, their contents lose their identity, and condense to homogeneous or heterogeneous residual bodies. These are returned to the distal surface of the cells, and then voided into the intestinal lumen. At the same time, synthesis and accumulation of numerous lipid droplets occurs, probably as a final product resulting from metabolism of the digested material. When feeding is over, the phagocytic cells are filled with lipid droplets, acquiring their typical appearance.It is suggested that disintegration of phagocytic cells during starvation is balanced by proliferation and differentiation of neoblasts into new phagocytic cells during the feeding-starvation cycle.     

不同生殖期鳜肝脏超微结构变化的观察

应用透射电镜对生殖季节与非生殖季节鳜肝脏超微结构的变化进行了观察。鳜肝细胞含有单个卵圆形的核,核仁清楚;细胞质内含有粗面内质网、线粒体、糖原颗粒和脂滴等细胞器和内含物。胆小管由相邻的数个肝细胞质膜凹陷围成,而肝血窦则由内皮细胞的胞质构成。还发现了贮脂细胞、枯否氏细胞和成纤维细胞。胆小管腔和窦周隙内浸润许多由肝细胞发出的微绒毛结构。鳜肝细胞的超微结构在产卵前后呈现明显变化：产卵前的肝细胞内富含线粒体、糖原颗粒和脂滴,粗面内质网发达;而产卵后的肝细胞内核仁发生迁移,部分细胞核囊泡化,糖原颗粒和脂滴排空,少数肝细胞具双核结构。非生殖期多数肝细胞核含有双核仁结构,胞质内溶酶体数量增多。     

Ultrastructural cytochemistry of the sex pheromone glands of Lutzomyia cruzi male sand flies (Diptera: Psychodidae: Phlebotominae)

The sex pheromone glands of Lutzomyia cruzi male sand flies (Diptera: Psychodidae) were analyzed by cytochemical techniques. In adult males, the epithelium at the fourth abdominal tergite is modified into a glandular epithelium, with large columnar gland cells located side by side. The gland cell cytoplasm contains a large number of mitochondria and peroxisomes, the latter with positive (electron-dense) reaction for catalase, a typical peroxisomal enzyme marker. The gland cell cytoplasm also contains a central vacuolated area, with a large number of electron-lucent vacuoles, not limited by a unit membrane. In well-preserved preparations such vacuoles present a homogenous and slightly electron-dense content, typical of lipid droplets. Indeed, incubation of the tergites with imidazole-buffered osmium tetroxide (to detect lipids) resulted in positive reaction in these vacuoles, as well as in between the microvilli of the gland cells. Use of the osmium–potassium iodide (Os–KI) technique allowed to demonstrate the presence of several endoplasmic reticulum (ER) profiles, as expected in secretory cells. Our data suggest that ER, lipid droplets and peroxisomes are involved in the sand fly pheromone biosynthesis.     

Mobilization and recovery of energy stores in traíra, Hoplias malabaricus Bloch (Teleostei, Erythrinidae) during long-term starvation and after re-feeding

In some neotropical environments, fishes often experience periods of poor food supply, especially due to extreme fluctuations in rainfall regime. The fish species that experience periods of drought such as the traíra Hoplias malabaricus (Bloch 1794), may stand up to long-term food deprivation. In this study, experiments were performed in order to determine the dynamic of utilization of endogenous reserves in this species during starvation. Adult traíra were both fasted for 30–240 days and re-fed for 30 days following 90 and 240 days of fasting. Glycogen and perivisceral fat were primary energy substrates consumed. During the first 30 days, fish consumed hepatic and muscular glycogen, without exhausting these reserves, and used lipids from perivisceral fat. Hepatic lipids were an important energy source during the first 60 days of starvation and perivisceral fat were consumed gradually, being exhausted after 180 days. Protein mobilization was noticeable after 60 days of fasting, and became the major energy source as the lipid reserves were decreased (between 90 and 180 days). Following the longest periods of food deprivation, fish had utilized hepatic glycogen again. Fish re-fed for 30 days after 90 and 240 days of fasting were able to recover hepatic glycogen stores, but not the other energy reserves.     

Changes in plasma thyroxine,triiodothyronine and cortisol associated with starvation in rainbow trout (Salmo gairdneri)

Synopsis Chronically starved rainbow trout (Salmo gairdneri) showed a significant fall in liver size, total liver glycogen, liver glycogen concentration and plasma glucose levels. Liver lipid concentration did not differ significantly from controls although total liver lipid reserves fell during the first 40 days of starvation but had partly recovered after 65 days of starvation. Plasma cortisol and T3 levels did not show consistent changes concomitant with food deprivation over the 65 day period of the experiment. However, plasma T4 levels in fish starved for 40 or 65 days were significantly lower than comparably fed animals. The involvement of T4 in intermediate metabolic processes in salmonids is discussed.     

Destructive and restorative processes of the liver in rats under intensive physical loading

The ultrastructure of rat liver cells after running exercise was investigated. When rats were trained for a month and sacrificed immediately after the last exercise it was revealed that the number of liver cells mitochondria increased, but many of them had alterations: mitochondria became swollen, had lucid matrix. There were some variations in degree of alterations between different mitochondria: a) in the same hepatocyte, b) in different hepatocytes of the same animal, that was connected with individual sensitivity of organelles on the levels of the cell and of the organ. Rough endoplasmic reticulum bore few ribosomes. Glycogen was absent. There were abundant vesicles of smooth endoplasmic reticulum, autophagic vacuoles and peroxisomes in the liver cell cytoplasm. Adaptation of rat liver to the exercise programme becomes evident by 1.5 month of exercise. Mitochondria and rough endoplasmic reticulum were numerous and of normal structure. There were many peroxisomes and glycogen granules in the cytoplasm of hepatocyte. The presence of large autophagic vacuoles in the cytoplasm of some hepatocytes were obviously connected with more rapid destruction of some organelles, than in control.     

Fine Structure of the Fat Body and its Bacteroids in Blattella germanica (Blattodea)

The abdominal fat body of the cockroach Blattella germanica contains three characteristic cell types—trophocytes, bacteriocytes and urate cells—which have been investigated by electron microscopy. The trophocytes are rich in lipid droplets of different sizes; glycogen, rough endoplasmic reticulum and mitochondria are also abundant. In females immediately after eclosion, the trophocytes contain a greater number of lipid droplets, some of which have different electron density; glycogen and cytoplasmic organelles are clearly reduced. The bacteriocytes hold rod-like and spherical bacteroids, which are encapsulated by a vacuolar membrane; they show a thin cytoplasmic membrane and an evident cell wall surrounded by a membrane-like outer envelope. The bacteroids appear to be dividing either by transverse partition or by budding. The urate cells, adjacent to the bacteriocytes, are characterized by complex urate vacuoles delimited by a double layer-structure.     

The regulation of endogeneous energy stores during starvation and refeeding in the somatic tissues of the golden perch

Adult golden perch Macquaria ambigua were fed to satiety, starved for up to 210 days, or starved for 150 days then fed to satiety for 60 days to investigate the utilization of energy stores in response to food deprivation and re-feeding. Golden perch sequentially mobilize energy from hepatic tissue, extra-hepatic lipid, and finally muscle components in response to food deprivation. The relative size of the liver was significantly reduced by 30 days after the onset of food deprivation due to the simultaneous mobilization of lipid, protein and glycogen reserves. These stores were renewed rapidly within 30 days by satiety feeding. Mobilization of lipid stores in perivisceral fat bodies occurred between 30 and 60 days of food deprivation. These deposits were also renewed upon re-feeding, although not as rapidly as liver reserves. The glycogen content of the epaxial muscle was reduced by the 60th day of food deprivation but subsequently increased indicating the mobilization of other energy reserves. The concentration of muscle lipid decreased after 90 days of food deprivation. The only significant response in body composition observed in the fish fed to satiety throughout the study was an increase in the relative size of the perivisceral fat bodies. The results of this study suggest that golden perch are well adapted to cope with extended periods of food deprivation, storing energy as perivisceral fat when food is readily available and having a clearly sequential process for mobilizing energy when food is scarce which largely protects the integrity of the musculature.