Action of metabolic hormones on the fine structure of rat liver cells III. Effects of adrenalectomy and administration of cortisone

Electron microscopic studies were made of hepatocytes from sham-operated rats, adrenalectomized animals fasted 15 hours, and adrenalectomized rats fasted 15 hours but given a single I.P. injection (10 mg) of cortisone acetate. The objective of this work was to define the earliest morphological response of hepatocytes to injection of a glucocorticoid and to provide additional information on the mechanism of hormone action at the cellular level. Hepatocytes from fasted, adrenalectomized rats contained no glycogen particles and very little smooth endoplasmic reticulum (SER). In addition the rough endoplasmic reticulum was disorganized and showed fewer ribosomes and polysomes than found in liver cells from sham-operated rats. Two hours after glucocorticoid injection glycogen particles were seen in numerous centrilobular cells and some periportal hepatocytes. Elements of SER were associated with the glycogen particles. By 4 hours after hormone injection abundant glycogen was found in all hepatocytes. Centrilobular cells showed dispersed glycogen with extensive tubules of SER associated with the glycogen particles. Periportal hepatocytes accumulated glycogen as dense masses scattered throughout the cytosome. SER occurred mainly at the edges of the glycogen masses. Midlobular cells showed glycogen patterns intermediate between periportal and centrilobular cells; masses of dispersed glycogen with abundant SER occurred within and around the glycogen areas of the cells. Glucocorticoid stimulation also caused cisternae of RER to align in parallel arrays, and more ribosomes and polysomes appeared on membranes of RER than in similar cells from adrenalectomized rats. The interpretation is offered that the glucocorticoid-stimulated proliferation of SER is the morphological expression of induced microsomal enzyme synthesis (glucose-6-phosphatase) known to occur under these hormonal conditions.     

Fine structure of hepatocytes from fasted and fed rats

The fine structure of hepatocytes from rats maintained on a controlled feeding schedule are described. Liver samples were processed for electron microscopy, histochemistry and chemical determinations of glycogen at precise time-intervals following a 30-hour fast and a 2-hour meal. Hepatocytes from 30-hour-fasted rats with extremely low hepatic glycogen levels were devoid of glycogen particles. Centrilobular cells showed areas of the cytoplasm rich in vesicles of smooth endoplasmic reticulum (SER) while periportal hepatocytes contained less extensive regions of SER. Soon after feeding the fasted rats, glycogen particles appeared in regions of the cell rich in SER. Centrilobular hepatocytes contained numerous glycogen areas which were infiltrated with tubules of SER, while periportal cells showed dense glycogen deposits with SER restricted to the periphery of the masses of glycogen. Throughout glycogen deposition each glycogen particle was closely associated with membranes of SER until maximum glycogen deposition was achieved 12 hours after initiation of feeding. At this point SER was reduced to the lowest amounts of the time-periods studied. During stages of glycogen depletion SER proliferated and reached the highest concentration measured in this study. Tubules of SER were present throughout the glycogen masses of centrilobular hepatocytes, whereas in periportal cells the organelle was restricted to the periphery of the glycogen masses. It is concluded that SER is associated with glycogen particles in rat hepatocytes during both deposition and depletion of glycogen.     

Glycogen metabolism in cultured chick hepatocytes: A morphological study

Ultrastructural and autoradiographic observations of cultured chick hepatocytes under the following conditins are described: Induction of glycogen synthesis with glucose alone and glucose plus insulin, and glucagon-induced glucogen glucogen breakdown. Profiles of hepatocytes cultured in medium containig 10 mM glucose showed typical cellular organelles and occasionally a few glycogen granules. After incubation of hepatocytes with ³H-glucose, silver grains were found over these sparse glycogen granules, indicating a low level of glycogen synthesis by a few cells. After addition of 75 mM glucose for 1 hr about 3% of the profiles of cells showed glycogen, and by 24 hr half of the hepatocytes had glycogen. Addition ofinsulin plus glucose induced glycogen synthesis in 82% of the cells after 6 hr. and by 24 hr almost every cellular profile showed glyocgen particles. Morphologically, glycogen accumulation was similar whether the cells were stimulated by high glucose or by glucose plus insulin: glycogen granules appeared in restricted regions of the cytoplasm, which were rich in smooth endoplasmic reticulum (SER), and peroxisomes were found close to the newly deposited glycogen particles. At maximum glycogen accumulation the association of SER and peroxisomes with glycogen was less obvious. Glycogenolysis induced by incubation of glycogen-rich hepatocytes with glucagon resulted in proliferation of SER in the glycogen regions of the cells. These observations are compatible with the concept of regions in the hepatocyte cytoplasm specialized for glycogen metabolism. Possible roles for SER and peroxisomes found near glycogen particles and other organelles in hepatic glycogen metabolism are discussed.     

Rapid alterations induced by insulin in hepatocyte ultrastructure and glycogen levels

The speed with which insulin alters hepatocyte ultrastructure and glycogen levels in insulin-deficient rats has been studied. Insulin deficiency was induced with alloxan, followed by insulin treatment with regular and NPH insulin. Rats were killed at various times after the insulin injection, blood samples were obtained, plasma glucose levels were determined, and liver samples were prepared for electron microscopy and glycogen determinations. Plasma glucose levels in insulin-deficient rats declined to normal values by 4 hours post insulin, returning to insulin-deficient levels by 8 hours post insulin. Hepatic glycogen was considerably reduced in the insulin-deficient rats. By 1 hour post insulin hepatic glycogen increased, reached maximal levels by 8 hours, then declined to insulin-deficient levels by 36 hours. The ultrastructural appearance of both centrilobular and periportal hepatocytes from insulin-deficient rats showed abundant vesicular smooth endoplasmic reticulum (SER), decreased rough endoplasmic reticulum (RER), and enlarged RER intracisternal spaces. One-half hour post insulin, centrilobular hepatocytes were unchanged. In periportal hepatocytes, however, vesicular SER was no longer visible, the RER intracisternal spaces appeared normal, and the amount of RER had increased. By 1 hour post insulin the centrilobular hepatocytes showed similar ultrastructural changes. These changes became more pronounced in the next few hours and remained through 24 hours. By 36 hours both centrilobular and periportal hepatocytes appeared similar to those in the insulin-deficient rat. These results demonstrate the rapid and lobular-specific effects insulin has on the hepatocyte.     

Lobular and cellular patterns of early hepatic glycogen deposition in the rat as observed by light and electron microscopic radioautography after injection of 3H-galactose

Very low hepatic glycogen levels are achieved by overnight fasting of adrenalectomized (ADX) rats. Subsequent injection of dexamethasone (DEX), a synthetic glucocorticoid, stimulates marked increases in glycogen synthesis. Using this system and injecting ³H-galactose as a glycogen precursor 1 hr prior to sacrifice, the intralobular and intracellular patterns of labeled glycogen deposition were studied by light (LM) and electron (EM) microscopic radioautography. LM radioautography revealed that 1 hr after DEX treatment, labeling patterns for both periportal and centrilobular hepatocytes resembled those in rats with no DEX treatment: 18% of the hepatocytes were unlabeled, and 82% showed light labeling. Two hours after treatment with DEX, 14% of the hepatocytes remained unlabeled, and 78% were lightly labeled; however, 8% of the cells, located randomly throughout the lobule, were intensely labeled. An increased number of heavily labeled cells (26%) appeared 3 hr after DEX treatment; and by 5 hr 91% of the hepatocytes were intensely labeled. Label over the periportal cells at this time was aggregated, whereas centrilobular cells displayed dispersed label. EM radioautographs showed that 2 to 3 hr after DEX injection initial labeling of hepatocytes, regardless of their intralobular location, occurred over foci of smooth endoplasmic reticulum (SER) and small electron-dense particles of presumptive glycogen, and in areas of SER and distinct glycogen particles. After 5 hrs of treatment with DEX, the intracellular distribution of label reflected the glycogen patterns characteristic of periportal or centrilobular regions.     

Fluoxymesterone-induced inclusion bodies in dog liver

A large dose of androgenic steroid fluoxymesterone (FM) was orally administered to beagle dogs for 15 days. At sequential time intervals, liver tissues were obtained by needle biopsy for light and electron microscopic examinations. The most prominent alteration of the hepatocytes was seen at 6 to 10 days of treatment in the endoplasmic reticulum. SER had proliferated. RER had lost their parallel arrangement with loss of ribosomes and the cisternae were elongated and meandering. Multilayers of concentrically arranged paired membranes, fingerprints, had enclosed the mitochondria, microbodies, lipids, and other cytoplasmic constituents. The paired membranes were in continuity with SER and RER at the periphery of the fingerprints. Horseshoe-shaped, incomplete rings or complex forms of multilayers of the paired membranes coexisted with fingerprints. Another characteristic structure was the presence of the elongated, bent endoplasmic reticulum aligned glycogen particles between the intercisternal spaces. It seemed that these cisternal structures fuse at their ends and become glycogen-bearing fingerprints which are transformed into tightly packed smooth glycogen-free ones. The fingerprints disappeared after cessation of FM administration. These results led to the conclusion that the FM-induced fingerprints originate from elements of the endoplasmic reticulum.     

Effect of cerium on the rat liver: an ultrastructural and biochemical study.

In rats, liver steatosis and necrosis were induced by cerous chloride (CeCl3) and the evolution of these changes was examined. By electron microscopy, 17 hours after CeCl3 treatment, dilation, disorganization and degranulation of the rough endoplasmic reticulum (RER) were noted with an increase in the number and electron density of lysosome-like bodies. In addition, nuclear chromatin showed showed a marked focal electron density, and the nuclear membrane appeared to be interrupted. At 24 hours, the RER was markedly dilated and degranulated, with free ribosomes aggregated in the cytoplasm. The Golgi cisternae appeared to be empty. There was an increase in the number and size of lipid droplets, with depletion of glycogen. At 48 hours, a massive proliferation of smooth endoplasmic reticulum (SER) vesicles occurred. Large lipid droplets were scattered throughout the cytoplasm, while the mitochondria displayed mild changes. By the 8th day, the number of lipid droplets returned to normal; no abnormalities were detected in the other cell organelles. Biochemically, the total hepatic ATP levels fell significantly by the 12th hour, dropping to a minimum by the 48th hour. The liver was gradually depleted of glycogen within the first 48 hours, while hepatic triglycerides increased rapidly, reaching a peak at 96 hours. Exogenous administration of adenine, ATP (adenosine triphosphate), or tryptophan completely prevented CeCl3-induced mortality; hepatic fat accumulation and necrosis were markedly decreased. Glucose, dl-methionine, and choline had no protective effect. It appears that a defect in hepatocellular lipoprotein synthesis and/or release may be responsible for lipid accumulation.     

Rat liver alterations after chronic treatment with hexachlorobenzene

Summary Groups of female rats were treated orally with 0.5, 2.0, 8.0, and 32 mg/kg hexachlorobenzene twice a week for 203 days. The liver content of hexachlorobenzene was found to be dose-dependent. In the animals treated with the highest dose the concentration was 273 g/g hexachlorobenzene. In the fresh and fixed hepatic tissue of the treated animals pink fluorescence was observed. Electron microscopy revealed a dose dependent enlargement of all hepatocytes due to proliferation of the SER in the centrolobular area or to increased glycogen deposits (- or -particles) and SER in the intermediary and periportal area. Numerous porphyrin deposits and siderosomes, intimate disorganisation and moderate dislocation of the RER and a moderate enlargement of bizarre-shaped mitochondria were recognized. The relationship between porphyrin crystals and mitochondria on the one hand and between SER and glycogen deposits on the other is discussed.G.K. is supported by the Deutsche ForschungsgemeinschaftThe authors are indebted to S. Vellguth and J. Seidel for their technical assistance.     

Neurofilament and glycogen changes during cold acclimation in the trochlear nucleus of lizards (Sceloporus undulatus).

In lizards (Sceloporus undulatus), long term (13 or 19 weeks) acclimation to an environment of 6 degrees C produces a striking increase in the argyrophilic neurofibrillar network in most large perikarya of the trochlear nucleus. In electron micrographs the cells contain numerous bundles of 10-30 regularly-spaced 90 A neurofilaments. In the cells from warm acclimated animals, a plexus of neurofibrils is seen by light microscopy. The electron micrographs show scattered neurofilaments and fewer, thinner bundles than in the cold. Within the cell bodies of the cold animals, glycogen particles are organized in regional accumulations from which other organelles are excluded except for the bundles of neurofilaments which are distributed throughout the cytoplasm. The aggregations of rough endoplasmic reticulum (RER) are also penetrated by the neurofilament bundles. The increased neurofilamentous network in the cold is not accompanied by obvious changes in the amount or distribution of RER or of microtubules which are present in limited numbers in both conditions. The dendrites of trochlear cells and axon terminals within the nucleus also show a cold induced increase in neurofilaments, as well as in the distinctive accumulations of glycogen particles.     

Medroxyprogesterone induced changes in rat liver. A light and electron microscopic study

Medroxyprogesterone acetate (MPA) induced morphological alterations in the normal female rat liver and after carbon tetrachloride exposure were assessed by light and electron microscopy and morphometry. Administration of MPA increased the smooth endoplasmic reticulum (SER) in normal rat. Carbon tetrachloride, injected intraperitoneally for four weeks, caused fatty accumulation, enlargement of the cell nucleus and decrease of the rough endoplasmic reticulum (RER) and increase of the SER. After one week of recovery, without treatment, the fatty infiltration and the SER membranes decreased and the RER increased. MPA, following injury, induced a decrease in the fatty accumulation and mitochondrial volume density and the hepatocyte nuclear and cytoplasm size normalized. The SER membrane volume density increased and the surface density decreased, the RER membrane volume density increased while the surface density did not alter significantly. The volume densities of the mitochondria and the peroxisomes decreased in the MPA treated CCl4 exposed groups. The results demonstrate that MPA induces a proliferation of the SER in normal rat liver and after hepatic injury the compound has a beneficial effect on the regeneration.     

Cocaine hepatotoxicity in mice: histologic and enzymatic studies

The severity of hepatotoxicity in CF-1 mice given 5 daily doses of 5, 10, and 20 mg cocaine/kg body weight and sacrificed 24 hr after the last injection appeared to be dose-dependent. Using light microscopy, the hallmark of cocaine early toxicity was manifested by pallor and ballooning of the hepatocytes in the midzonal and then in the centrilobular areas. Degeneration and necrosis of the liver cells in the same zones were encountered while the hepatocytes in the periportal areas remained intact. When examined under the electron microscope, such pallor and ballooning of the hepatocytes appeared to to be due to dilation of the rough endoplasmic reticulum (RER) profiles which often revealed a significant loss of their ribosomes. Dilation of the smooth endoplasmic reticulum (SER) was also common and moderate proliferation of peroxisomes was frequently seen. In the degenerating hepatocytes, the 2 forms of endoplasmic reticulum were difficult to recognize and the peroxisomes appeared sparse. Cocaine treatment elevated the level of glutamic pyruvic transaminase and glutamic oxaloacetic transaminase in a dose-dependent manner. Although hepatic cytochrome P450 was slightly increased in the low dose groups, a reduction in the enzyme activity was noticed in the group treated with 20 mg cocaine/kg. However, hepatic reduced glutathione content manifested a significant increase in the group which received 20 mg cocaine/kg.     

Action of metabolic hormones on the fine structure of liver cells. II. Effects of hypophysectomy and chronic administration of somatotropin

Electron microscopic studies of hepatocytes from sham-operated, hypophysectomized, and hypophysectomized rats treated with somatotropin were undertaken in order to obtain morphological information on the mechanism of hormone action at the cellular level. Fed and fasted animals from each of the above groups were studied. Hepatocytes from fed rats of the three groups showed similar characteristics: periportal cells contained large dense masses of glycogen throughout the cytosome with relatively little smooth endoplasmic reticulum; mid-lobular cells displayed small dense masses of glycogen with few associated elements of smooth endoplasmic reticulum; centrilobular hepatocytes showed dispersed glycogen and abundant smooth endoplasmic reticulum. Hepatocytes from hypophysectomized rats fasted fifteen hours showed significant alterations in structure when compared with liver cells from sham-operated animals. Differences noted were: decrease in mid-lobular cell size and amount of smooth endoplasmic reticulum; decrease in glycogen, number of ribosomes and polysomes per hepatocyte; disorganization of the rough endoplasmic reticulum; and swelling of mitochondria and increase in number per cytoplsmic volume. Periportal hepatocytes from fasted hypophysectomized rats treated with somatotropin for ten days contain numerous small masses of glycogen with elements of smooth endoplasmic reticulum at the periphery of the glycogen masses. Mid-lobular and centrilobular hepatocytes contain masses of dispersed glycogen with an abundance of smooth endoplasmic reticulum associated with the glycogen particles.     

Alterations in hepatic fine structure after chronic exposure of rats to dexamethasone

The objective of this study was to determine the effects of chronic dexamethasone (DEX) administration on hepatic ultrastructure and to correlate these changes with plasma lipoprotein levels. Electron microscopic studies were made of hepatocytes from male rats killed 1, 3 and 5 days after DEX (2 mg, twice per day) administration. Three days after treatment plasma lipoprotein levels were highest and hepatocytes contained regions of the cytosome rich in elements of the smooth endoplasmic reticulum (SER). Osmiophilic particles were present in the tubules and vesicles of the SER, in the saccules and vacuoles of the Golgi complex, in secretory vesicles near the cell surface and in the space of Disse. DEX treatments also caused hepatocytes to accumulate tightly packed masses of β-particles of glycogen in some regions of the cell while other areas displayed dispersed glycogen particles that were associated with the SER. These observations are consistent with the hypothesis that glucocorticoids 1. cause an elevation of plasma lipoprotein levels by in creasing hepatic synthesis and secretion of VLDL, which involves the sequential participation of the ER, the Golgi complex and exocytosis of VLDL-containing vacuoles into the space of Disse, and 2. produce a change in the nature of the association of glycogen particles with the SER membranes in response to the physiological state of the animal.     

Effects of glucagon on hepatic glycogen and smooth endoplasmic reticulum

The action of glucagon on hepatic glycogen and smooth endoplasmic reticulum (SER) was studied in samples of liver taken sequentially from anesthetized rats. The physiological state of each animal was assessed by continuously monitoring aortic blood pressure and blood lactate/pyruvate ratios. High hepatic glycogen levels were established by using 10–12 hr fasted control-fed rats infused continuously with glucose. In rats receiving glucose only, hepatic glycogen levels remained above 5.0% during the 4-hr period of glucose administration. Centrilobular hepatocytes displayed an abundance of glycogen which often appeared dispersed with elements of SER between the glycogen particles. Periportal cells had dense clumps of glycogen with few vesicles of SER restricted to the periphery of the glycogen masses. The addition of glucagon to the glucose infusate caused a marked stimulation of glycogenolysis. In these rats, the hepatic glycogen level (x?±SE) was 6.71±.15% 1 hr after glucose and declined after initiation of glucagon infusion as follows: 5.86±.29% (15 min), 4.89±.26% (1 hr), 2.16±.40% (2 hr), and 1.66±.29% (3 hr). The fine structure of hepatocytes showed a dramatic response to the administration of glucagon. The glycogen regions of the cells were noticeably decreased in size and number of glycogen granules 3 hr after initiation of glucagon infusion, and SER was abundant in both periportal and centrilobular hepatocytes. The interpretation offered is that glucagon induces the formation of new SER membranes which participate in glycogen breakdown and/or glucose release from hepatocytes.     

Alterations in hepatic fine structure after chronic exposure of rats to dexamethasone.

The objective of this study was to determine the effects of chronic dexamethasone (DEX) administration on hepatic ultrastructure and to correlate these changes with plasma lipoprotein levels. Electron microscopic studies were made of hepatocytes from male rats killed 1, 3 and 5 days after DEX (2 mg, twice per day) administration. Three days after treatment plasma lipoprotein levels were highest and hepatocytes contained regions of the cytosome rich in elements of the smooth endoplasmic reticulum (SER). Osmiophilic particles were present in the tubules and vesicles of the SER, in the saccules and vacuoles of the Golgi complex, in secretory vesicles near the cell surface and in the space of Disse. DEX treatments also caused hepatocytes to accumulate tightly packed masses of beta-particles of glycogen in some regions of the cell while other areas displayed dispersed glycogen particles that were associated with the SER. These observations are consistent with the hypothesis that glucocorticoids 1. cause an elevation of plasma lipoprotein levels by increasing hepatic synthesis and secretion of VLDL, which involves the sequential participation of the ER, the Golgi complex and exocytosis of VLDL-containing vacuoles into the space of Disse, and 2. produce a change in the nature of the association of glycogen particles with the SER membranes in response to the physiological state of the animal.     

Chlordecone-induced potentiation of carbon tetrachloride hepatotoxicity: a light and electron microscopic study

Previous studies have shown that a chlorinated pesticide, chlordecone (Kepone), greatly potentiates carbon tetrachloride (CCl4) hepatotoxicity and lethality (Curtis, L.R., Williams, W.L., and Mehendale, H.M. (1979). Toxicol. Appl. Pharmacol. 51, 283-293; Curtis, L.R., and Mehendale, H.M. (1980). Drug Metab. Dispos. 8, 23-27). The present study describes sequential morphologic changes which occurred in livers of rats given a "nontoxic" level of chlordecone (10 ppm for 15 days) followed by a single injection of CCl4 (0.1 ml/kg). The hepatic alterations were examined 1 to 36 hr after exposure of the rats to CCl4. Those changes were compared to hepatic alterations which occurred in rats that received the same dose of chlordecone (10 ppm for 15 days) or a single injection of CClr (0.1 ml/kg) alone. The only change noted in livers from rats that received chlordecone alone was focal increase in smooth endoplasmic reticulum (SER) of hepatocytes at 24 hr and continuing throughout the time course of the experiment. Livers from animals that received CCl4 alone showed morphologic changes at 6 hr consisting of glycogen loss, increase in SER, and dilatation of rough endoplasmic reticulum (RER) in pericentral hepatocytes. Accumulation of small lipid droplets was also noted in midzonal hepatocytes. After 6 hr, there was no further increase in severity of injury. At 12 hr recovery was noticeable and, by 36 hr, livers from the CCl4 group appeared normal. Prior administration of chlordecone greatly potentiated pathologic changes in livers of animals that received CCl4. By 4 hr, there was total loss of glycogen in hepatocytes throughout the entire lobule. Small lipid droplets were present in pericentral, midzonal and periportal hepatocytes. Hepatocytes with extremely dilated RER were randomly scattered throughout the entire lobule. At 6 hr, there was further accumulation of lipid in the form of large droplets in hepatocytes. Focal, necrotic cells surrounded by polymorphonuclear leukocytes were randomly distributed throughout the lobule. The number of necrotic foci had progressively increased at the 12- and 24-hr intervals. By 36 hr, confluent areas of necrosis in pericentral and midzonal areas were observed in livers of some animals. This study indicates that although the combination of chlordecone and CCl4 produces much greater hepatic injury resembling damage due to a massive dose of CCl4, histologically, some differences in the progression and distribution of hepatocellular damage within the lobular architecture of the liver are evident.     

Freeze fracture and scanning electron microscope studies on the nuclear envelope and perinuclear cytomembranes (parabasal apparatus) in the protozoan, Lophomonas blattarum

Phase contrast, scanning electron microscope (SEM), and freeze fracture studies on the parasitic flagellate, Lophomonas blattarum, have demonstrated that the endomembrane system (parabasal apparatus) is highly ordered, restricted in position to a perinuclear zone at the anterior end of the organism, and is supported in this localized cytoplasmic region by overlapping sheets or plates of microtubules, previously called the calyx and axial filament, which may participate in supporting the nucleus-endomembrane system in a restricted region of the cell. Light microscope observations, SEM and freeze fracture data provide support to previous views that the rough- and smooth-surfaced endoplasmic reticulum are interconnected and attached to the outer layer of the nuclear envelope. The continuity of these membrane systems provides an orderly and restricted packing in the perinuclear cytoplasm since other areas of the cell may become filled with yeast. These flagellates are especially adept at phagocytosis of entire yeast. In yeast-laden cells, the flagella-nucleus-parabasal body-calyx-axial filament complex may separate from the remainder of the cell and assume a motile existence for a time. The significance of the described relationships, in addition to providing efficiency in endomembrane localization, may also reflect synthesis of enzymes and proteins by the RER and packaging in the SER, both of which are continuous. Granules characteristic of glycogen are concentrated around the SER which may be involved in glycogen metabolism. Although critical information is lacking, the endomembrane system may also be involved in the synthesis and morphogenesis of lysosomes, and perhaps peroxisomes. Lophomonas thus amplifies a highly ordered spatial relationship between the nuclear envelope and the ER.     

Electron microscopical investigation on aldrin-induced hepatocyte pathology in Rana catesbeiana, with special emphasis on peroxisomes.

We examined the effect of aldrin on hepatocyte ultrastructure in liver of Rana catesbeiana. The frogs were experimentally exposed to chemical substance and liver fragments processed for routine transmission electron microscopy. Hepatic peroxisomes were visualized after incubation with alkaline 3,3'-diaminobezidine (DAB) method. Ultrastructural analysis revealed progressive hepatocyte changes induced by this drug. After 2-weeks, in the hepatocytes the nuclear envelop and the cisternae of both smooth and rough endoplasmic reticulum (SER und RER, respectively) were unusually enlarged. Reduction of glycogen granules associated with an increased frequency of lysosomes was observed. Normal appearing peroxisomes were present in clusters. Lipid droplets were also visualuzed. After 4-weeks, there was a new increase of glcogen associated with a great number of mitochondria and peroxisomes. Moreover, SER und RER were still dilated. Intracellular lipid inclusions became more abundant. These results suggest that the aldrin 250 induces ultrastructural changes in the hepatocyte of Rana catesbeiana.     

Circadian rhythm of hepatocellular structures in rats dosed phenobarbital

The circadian rhythm of liver weight and hepatic structures was studied in rats given a maximum tolerated dose of 70 or 80 mg/kg/day Phenobarbital during 7 days and killed at four different hours of the following day. The drug-induced increase in liver weight in % of body weight of rats was about 1% in the morning and in the afternoon in comparison with controls. At 6.00--7.00 during the glycogen maximum, the fine structure of hepatocytes was normal except enlargement and glycogen depletion of cells in the near neighbourhood of central veins. From 9.00--10.00 the mitotic activity was markidly increased and parallel to the progressive disappearance of glycogen the smooth endoplasmic reticulum (SER) became prominent. Hypertrophy of SER was fully developed at 15.00--16.00 and 17.00--18.00.     

Rubratoxin B hepatotoxicity: An electron microscopic study

Rubratoxin B, the principal toxin produced by Penicillium rubrum, causes hepatic damage. The present study describes ultrastructural changes in rat livers at intervals of 3, 6, 24, and 72 hr following administration of 0.36 mg/kg rubratoxin B. Three hours after treatment, minimal ultrastructural changes including loss of glycogen and dilatation of rough endoplasmic reticulum (RER) were observed in groups of hepatocytes. These groups of altered cells showed a random, multifocal pattern of distribution throughout the liver by light and electron microscopy. At 6 hr, hepatocytes in the focal areas affected by rubratoxin showed more severe ultrastructural changes. There was increased vesiculation of RER with loss of ribosomes from the membranes and disaggregation of polyribosome complexes. Mitochondria contained inclusions of cytoplasmic material and intramatrix myelin figures. Increased numbers of lipid droplets were observed in some cells. Sinusoids appeared to be slightly congested. At the 24-hr interval, hepatocytes in the affected areas showed progressive degenerative changes. Mitochondria were extremely swollen and showed loss of cristae and increased numbers of myelin figures in their matrix. Accumulation of cytoplasmic lipid was more evident, and autophagic vacuoles were occasionally seen. Nuclear changes in these hepatocytes included clumping and margination of chromatin. Sinusoids at the 24-hr interval were severely congested with blood cells, platelets, and fibrin. Endothelial cells lining the sinusoids were fragmented and, in some areas, completely absent. By 72 hr, most hepatocytes in the focal, involved areas were completely necrotic. Cytoplasmic organelles showed irreversible, degenerative changes and cell membranes were frequently ruptured. Sinusoids showed complete loss of lining endothelium, and the lumens were occluded with blood cells, fibrin, and cellular debris.