Early Ultrastructural Changes Research Articles

Glomerular ultrastructural lesions of idiopathic cutaneous and renal glomerular vasculopathy of greyhounds.

Idiopathic cutaneous and renal glomerular vasculopathy (CRGV) (Alabama rot) is a potentially fatal disease of unknown etiology that affects the skin and kidneys of racing- and training-age Greyhounds. Ultrastructural examinations were performed on two healthy control Greyhounds and 12 Greyhounds diagnosed with CRGV based on the presence of characteristic, well-demarcated cutaneous ulcers of the extremities (12/12), thrombocytopenia (< 200,000 platelets/dl) (12/12), and acute renal insufficiency (BUN > 40 mg/dl, serum creatinine > 2.0 mg/dl) (7/12). Early glomerular ultrastructural changes included endothelial swelling, detachment, and necrosis; membranous whorl formation; and platelet adhesion and aggregation. Some capillaries were occluded with aggregated platelets, cellular fragments, and fibrin. Later changes included narrowing of capillary lumina and thickening of glomerular capillary walls by subendothelial accumulation of flocculent, amorphous, variable electron-dense material and occasionally erythrocytes, cellular processes, and fibrin. Glomerular endothelial cells were increased in number and plump, with villouslike cytoplasmic projections. Mesangial cell cytoplasmic processes occasionally were interposed between the endothelium and the basement membrane. No etiologic agents or electron-dense deposits typical of immune complexes were observed. Although the specific etiology was not determined, the ultrastructural changes suggest that glomerular endothelial damage is an important early event in the pathogenesis of CRGV.

Correlation of alterations on Na(+)-K+/Mg+2 ATPase activity, lipid peroxidation and ultrastructural findings following experimental spinal cord injury with and without intravenous methylprednisolone treatment.

The sodium-potassium activated and magnesium dependent adenosine-5'-triphosphatase (Na(+)-K+/Mg+2 ATPase EC 3.6.1.3.) activity and lipid peroxidation and early ultrastructural findings are determined in rat spinal cord at the early stage of trauma produced by a surgical clip on the thoracal 2-7 segments. The effect of treatment with intravenous methylprednisolone (MP) was evaluated the basis of these biochemical alterations and ultrastructural findings in the same model. The specific activity of the membrane bound enzyme Na(+)-K+/Mg+2 ATPase was promptly reduced in as early as ten minutes following spinal cord injury and remained at a level lower than the levels in the control group and in the sham-operated group. Methylprednisolone treatment immediately after the trauma attenuated the inactivation of Na(+)-K+/Mg+2 ATPase. On the other hand, there was significant difference in lipid peroxide content between the sham-operated and the injured animals. Methylprednisolone treatment reduced thiobarbituric acid reactive substance (TBARS) content in Group IV. We determined a positive relationship among membrane-bound enzyme Na+K+/Mg+2 ATPase activity, malondialdehyde (MDA) content and early ultrastructural changes in the traumatized and treated groups.

Early ultrastructural changes in blood-brain barrier vessels of the rat embryo

The blood-brain barrier (BBB) in fetal rat brain has been shown by others to be more permeable to a variety of blood-borne solutes than the BBB in adults. We used ultrastructural morphometric methods to measured the density of putative vascular pores between the ages of embryonic day (E) 11 and birth to determine the structural basis for this relatively high permeability. We found that fenestrations, that are frequent at E11, declined rapidly and were last seen at E13 in intraparenchymal vessels and at E17 in pial vessels. Interendothelial junctions in fetal brain contained expanded clefts suggestive of paracellular channels at all ages examined, although they disappear after birth. Both of these features likely contribute to high fetal BBB permeability, but endothelial vesicles probably do not. The central nervous system is vascularized by ingrowth of capillary sprouts from the perineurial vascular plexus. Invading capillaries express BBB features in response to inductive signals from the surrounding neural tissue. We compared early ultrastructural changes in perineurial vessels, which are separated from neural tissue by a sizeable perivascular space, with those in intraneural vessels, which are totally enveloped by neural tissue, to determine whether the inductive interaction requires close cellular contact. For the most part, the perineurial and intraneural vessels matured in parallel. Furthermore, cerebellar vessels developed in parallel with cerebral vessels, even though they did not invade neural tissue until a comparatively late stage. These results suggest that intimate contact between neural tissue and vessel walls is not a requirement for BBB expression.

Early ultrastructural changes of thymocytes in T-2 toxicated mice.

Subcellular changes of mouse thymocytes after T-2 intoxication were observed for up to 48 hours after injection (HAI). The following changes were observed mainly in cortical lymphocytes at 8 and 16HAI: (1) nuclear projections, (2) nuclear macrocleft, and (3) characteristic features of apoptosis. Conclusively, it is suggested that apoptosis is partially involved in the early development of thymic lesions by T-2 toxin.

Morphogenesis of a zone-specific adrenocortical cytotoxicity in guinea pigs administered PD 132301-2, an inhibitor of acyl-CoA:cholesterol acyltransferase.

PD 132301-2, a novel inhibitor of acyl-CoA:cholesterol acyltransferase, is adrenotoxic to several laboratory animal species. Morphogenesis of a zona fasciculata-specific cytotoxicity was evaluated in male Hartley guinea pigs administered 100 mg/kg of PD 132301-2 for up to 7 days. Reversibility of adrenal effects was assessed after a 14-day drug withdrawal period (day 21). Serum cortisol concentrations were determined under basal conditions and after administration of adrenocorticotrophic hormone (ACTH) on days 1, 2, 4, 7, and 21. Isolated foci of cortical cell degeneration and necrosis were apparent in outer zona fasciculata by 2 hr and throughout the zona fasciculata at 6 hr. Early degenerative ultrastructural changes included aggregation of smooth endoplasmic reticulum (SER), variable condensation of mitochondrial matrices and swelling of cristae, partitioning of organelles, autophagosome formation, and disruption of lipid globules. Lesions progressed to locally extensive or diffuse zonal necrosis on days 1 and 2 and near complete ablation of zona fasciculata by day 4. Fasciculata cells remaining on day 4 had reduced numbers and increased size of lipid globules, increased lysosomes, and, occasionally, aggregates of SER and mitochondria. On day 7, SER proliferation and lipid depletion were apparent in remaining cells. ACTH responses were attenuated 24 hr after the first dose, and reduction in basal cortisol levels were seen by 24 hr after the second dose with both effects maximal on day 7. After a 14-day withdrawal period, ACTH responses and adrenal morphology returned to normal. It was concluded that PD 132301-2 induced rapid, reversible, zone-specific, morphologic, and functional adrenocortical effects. Furthermore, mitochondria and SER represented early subcellular targets of toxicity.

Early histological and ultrastructural changes in medullary fracture callus: Brighton CT, Hunt RM. J Bone Joint Surg [Am]73:832, 1991

Early histological and ultrastructural changes in medullary fracture callus: Brighton CT, Hunt RM. J Bone Joint Surg [Am]73:832, 1991

Early ultrastructural changes of surface epithelial cells isolated from rat gastric mucosa after exposure to ethanol with or without 16,16-dimethylprostaglandin E2.

Early ultrastructural changes of surface epithelial cells isolated from rat gastric mucosa caused by 15% ethanol were studied. The effect of 16,16-dimethylprostaglandin E2 (16,16-dimethyl-PGE2) on these changes was also examined. Findings by transmission electron microscopy showed ballooning and decreased density of mitochondria in addition to loss of microvilli and partial disruption of the surface cell membrane in cells exposed to 15% ethanol, although the nucleus looked normal. These ultrastructures were well preserved even after exposure to 15% ethanol when the cells were treated with 10(-6) M 16,16-dimethyl-PGE2 beforehand. These results may indicate that the surface cell membrane and mitochondria are the major targets for early injury by ethanol and protection by 16,16-dimethyl-PGE2 in surface epithelial cells. The mechanism by which 16,16-dimethyl-PGE2 prevents the damage is not known.

Ultrastructural changes in the hippocampal CA1 region following transient cerebral ischemia: evidence against programmed cell death

The ultrastructural changes in the pyramidal neurons of the CA1 region of the hippocampus were studied 6 h, 24 h, 48 h, and 72 h following a transient 10 min period of cerebral ischemia induced by common carotid occlusion combined with hypotension. The pyramidal neurons showed delayed neuronal death (DND), i.e. at 24 h and 48 h postischemia few structural alterations were noted in the light microscope, while at 72 h extensive neuronal degeneration was apparent. The most prominent early ultrastructural changes were polysome disaggregation, and the appearance of electron-dense fluffy dark material associated with tubular saccules. Mitochondria and nuclear elements appeared intact until frank neuronal degeneration. The dark material accumulated with extended periods of recirculation in soma and in the main trunks of proximal dendrites, often beneath the plasma membrane, less frequently in the distal dendrites and seldom in spines. Protein synthesis inhibitors (anisomycin, cycloheximide) and an RNA synthesis inhibitor (actinomycin D), administered by intrahippocampal injections or subcutaneously, did not mitigate neuronal damage. Therefore, DND is probably not apoptosis or a form of programmed cell death. We propose that the dark material accumulating in the postischemic period represents protein complexes, possibly aggregates of proteins or internalized plasma membrane fragments, which may disrupt vital cellular structure and functions, leading to cell death.

CORONARY ARTERY ULTRASTRUCTURAL CHANGES IN CARDIAC TRANSPLANT ATHEROSCLEROSIS IN THE RABBIT

Accelerated coronary atherosclerosis is the limiting factor for long-term survival of cardiac transplant recipients, but its pathogenesis is poorly understood. Morphologic and ultrastructural changes in suitable models may help explain the underlying mechanisms. In this study, early (3, 7, 14, and 21 days) and late (42 days) ultrastructural changes of the coronary artery were characterized in rabbit cardiac allografts. Thirty-four New Zealand white male rabbits (3.0-4.0 kg) served as donors and recipients. All recipients received cyclosporine (10 mg/kg/day i.m.) as immunosuppressant. In order to increase the normally very low cholesterol levels in rabbits, both the donor and recipient animals were fed a 0.5% cholesterol diet. Recipient animals were sacrificed between 3 days and 6 weeks after transplantation. The specimens from both donor and recipient were examined by transmission electron microscopy, immunohistochemistry, and morphometry. Our data indicate that intimal thickening was initiated with smooth muscle cell migration within 1 week after transplantation, and occurrence of macrophage-derived foam cells and vacuolized smooth muscle cells 3 weeks after transplantation. These changes occurred in the presence of an ultrastructurally intact endothelium. Platelets were only seldom seen adhering to the endothelium. In contrast, lymphocytes and monocytes were frequently found adhering to the endothelium at 2 and 3 weeks posttransplantation. From 3 weeks posttransplantation, lymphocytes were seen only occasionally in the intima but not in the media. This study suggests that early events elicit a change in the smooth muscle cells in the media to the secretory phenotype that migrates to the intima and proliferate. Lymphocyte and monocyte adhesion to the endothelium may enhance smooth muscle migration and proliferation. The large macrophage involvement may relate to the high serum cholesterol levels induced by the cholesterol diet. All these changes occurred in the presence of a structurally normal endothelium and without apparent platelet involvement.

Electron microscopic changes on the DMBA induced oral precancerous and cancerous lesion in hamster cheek pouch

Experiments were performed to study the early and late ultrastructural changes during hamster cheek pouch carcinogenesis using a regimen of topical application of 9, 10 dimethyl-1-2 benzanthracene (DMBA) twice a week in liquid paraffin oil. The DMBA was administered for a period of 2 and 4 1 2 months. Hamsters exposed to DMBA for 2 months developed moderate precancerous changes, whereas the hamsters treated with DMBA for 4 1 2 months developed frank and multiple oral tumors with a cauliflower appearance. The ultrastructural pathological changes seen were considerably increased at 4 1 2 months compared with a 2 month period of DMBA treatment. Untreated and solvant control hamsters cheek pouch treated for 2 and 4 1 4 months with liquid paraffin oil alone did not show any premalignant or malignant changes during this period.

An Improved Fixation Method for Transmission Electron Microscopy for the Histological Study of the Lens

An improved fixation technique for transmission electron microscopic observation that enables good fixation of all areas of the rat lens was devised. Immersion fixation with 0.1 M phosphate-buffered 1.25% glutaraldehyde at 4 degrees C for 12 hours followed by postosmication produced good results for all areas of the lens--anterior, equatorial, and posterior zones. The technique was particularly suitable for maintenance of the shape of the lens since practically no irregularity, vacuolar degeneration, or expansion of the intercellular spaces was noted. This technique, which requires only a relatively short time, was also useful for the detection of early ultrastructural changes associated with cataract in spontaneously diabetic WBN/Kob rats. We anticipate that our procedure will be widely applied.

Early acinar cell changes in caerulein-induced interstitial acute pancreatitis in the rat

Early ultrastructural and immunohistochemical changes caused by supramaximal secretory stimulation with caerulein were studied in the rat pancreas. The morphological basis for the earlier reported decrease of pancreatic juice secretion after supramaximal caerulein was the appearance of swollen and irregular zymogen-like material containing structures with short segments of budding bristle-coated membranes in the apical parts of acinar cells. Images of exocytosis of zymogen granules were only few. Later, marked vacuolization and signs of autophagocytosis are seen in the basal cytoplasm. Immunohistochemistry showed that the large zymogen containing structures were intensively labelled for trypsin at the early stages of the experiment (4-30 min). Later (1-2 h), the vacuoles were empty or contained occasional, small-labelled granules only. The pancreozymin-receptor antagonist proglumide as well as cycloleucine that inhibits protein synthesis by inhibiting the synthesis of S-adenosylmethionine, effectively prevented the caerulein induced acinar cell changes. The irregular zymogen containing structures with coated pits on their surface indicate disturbed zymogen granule formation leading to the accumulation of large lakes of zymogen material and finally to marked autophagocytosis in acinar cells. The effects of caerulein are receptor-mediated and depend on the process of methylation in the formation of zymogen granules.

Comparative endotoxin‐induced hepatic injury in young and aged rats

Recent studies have demonstrated that aged rats are more susceptible to the lethal effects of endotoxin (ET) than young rats. The early (15 min to 7 h) hepatic ultrastructural and biochemical changes induced by ET in young (6 months) and aged (24 months) rats were evaluated to elucidate cell populations and/or the mechanisms that may be responsible for the previously observed differential effects. Aged rats given ET had significantly increased numbers of neutrophils in hepatic sinusoids at 30 min and thereafter as compared with ET-treated young rats. Morphologic evidence of coagulation within hepatic sinusoids, including aggregates of fibrin enmeshed among polymorphonuclear leukocytes and platelet aggregates, was frequently observed in ET-treated aged rats but not in ET-treated young rats. In contrast, Kupffer cells of ET-treated young rats frequently contained phagocytized neutrophils and platelets, whereas this phenomenon was rarely observed in Kupffer cells of ET-treated aged rats. Hepatocellular morphologic injury was more pronounced and occurred at earlier time periods in ET-treated aged rats, and was accompanied by significant increase in hepatic transaminases. ET-treated aged rats had an earlier onset and greater severity of endothelial cell injury than did ET-treated young rats. The results of this study indicate a greater aggregation of blood elements in the hepatic sinusoids of aged rats following the intravenous administration of ET, which suggests that a greater diminution in microcirculation was induced in aged rats by ET. Additionally, the increased phagocytosis of inflammatory cells by Kupffer cells of young rats may be a mechanism which affords protection against endotoxin-induced lethality.

Canine X-linked muscular dystrophy: Morphologic lesions

Gross pathologic lesions and light microscopic and ultrastructural features of skeletal muscle lesions in canine X-linked muscular dystrophy (CXMD) were studied in dogs from 3 months to 6 years of age. Necrosis and regeneration were present at all ages, but were most prominent in the youngest dogs studied. Increased intracytoplasmic calcium, as evidenced by positive alizarin red S staining, was associated with fiber necrosis, but was also seen in small numbers of otherwise normal fibers. Progressive changes included development of severe fiber size variation, endomysial and perimysial fibrosis, prominent cytoplasmic disorganization, internalization of myonuclei, mitochondrial proliferation, mild fat infiltration, and alterations in the fiber-type pattern. The most consistent early ultrastructural changes were dilatation of the sarcoplasmic reticulum and focal subsarcolemmal areas of degeneration. Convincing sarcolemmal defects were not found. Z-band streaming was present at all ages, and Z-band duplication and nemaline rods were seen in older dogs. Evidence for abnormal regeneration was found in the oldest dog, and was associated with extensive fibrosis. These findings document the progression of lesions in CXMD, and illustrate the profound alterations in fiber organization and fiber type that may occur in late stages of dystrophin-deficient muscular dystrophy.

Early ultrastructural changes in skeletal muscle biopsies from patients with mitochondrial myopathy

Early ultrastructural changes in skeletal muscle biopsies from patients with mitochondrial myopathy

Field emission scanning electron microscopic study on the noninfarcted myocardium after myocardial infarction.

Intracellular membranous ultrastructural changes in the noninfarcted myocardium of the left ventricle were studied by examining experimental myocardial infarction in rats with a field emission scanning electron microscope. The left coronary artery was ligated at its origin for 7 days. The resulting infarcts comprised approximately 40% of the total area of the left ventricles. In a light microscopic study, the transverse diameter of myocytes had increased to 17.1 +/- 2.9 microns in the noninfarcted myocardium, compared to 14.1 +/- 3.1 microns in the myocardium of sham-operated rats (p less than 0.01). For the scanning electron microscopic study, the specimens were processed according to the Osmium-DMSO-Osmium method. Marked morphological changes of intracellular membranous structures were found in the noninfarcted myocardium, especially in the lacelike network of sarcoplasmic reticulum which occasionally formed large flattened cisternae, and in the marked proliferation of surface caveolae. It is speculated that these early ultrastructural changes in the noninfarcted myocardium reflect an adaptation of microorganellae in the failing hearts of the rats.

Mycelium-yeast transition in Histoplasma capsulatum. Early ultrastructural changes.

The ultrastructural changes which occur during the first 24 h of mycelium to yeast transition have been studied in the dimorphic fungus Histoplasma capsulatum. A temperature shift controls mycelial to yeast transition. During the first 24 h respiratory rate, ATP and cytochrome concentration fall to very low levels. Ultrastructural observations showed that the plasma membrane became undulated and the cell wall lost its characteristic fibrous outer layer. At 8 h the ordered lamellar structure of the mitochondria was no longer apparent. 24 h after the temperature shift 70% of the cells were lysed. The remaining cells contained many cytoplasmic membrane structures; mitochondria were rarely observed. These changes are considered to be the morphological expression of the physiological events characteristic of stage one in mycelial to yeast transition.

Mode of Action of Itraconazole: Morphological Aspects

The broad spectrum of antifungal activity of itraconazole is verified by morphologic criteria at the light and electron microscopical level. Yeast and fungal species examined are Candida albicans, Cryptococcus neoformans, Paracoccidioides brasiliensis, Sporothrix schenckii, Pityrosporum ovale, Trichophyton rubrum and Aspergillus fumigatus. Exposures of cultures of these yeasts and fungi to itraconazole results in dose- and time-dependent alterations which vary in nature and intensity from one species to another. The most striking gross morphological change is seen in the biphasic species and consists of the abolishment of morphogenic development of the blastospore into the hyphal forms (C. albicans and P. ovale) and of the hyphal into the yeast forms (P. brasiliensis). Furthermore, the outgrowth and development of inoculated A. fumigatus hyphae into sporulating vesicles is almost completely abolished. The above mentioned effects on morphogenesis are achieved in the 10(-10)-10(-7) M range. Except for P. ovale, the earliest ultrastructural changes after itraconazole consist of abnormalities at the plasma membrane, the cell wall and cytoplasmic vacuoles. They precede a marked increase in cell volume, defective cell division, abortive hyphal outgrowth and loss of cell viability. These changes are identical to those previously described after miconazole and ketoconazole treatment. However, when compared to the other available azole-derivatives sharing the same basic mechanisms of action, itraconazole displays an exceptionally strong activity against A. fumigatus which can be morphologically translated by a potent necrotizing action on hyphae and inhibition of vesicle formation and sporulation. The in vitro effects of itraconazole are supported by data obtained from microscopic examinations of samples derived from patients with experimental animals infected with various fungal organisms.

Early ultrastructural changes after low-dose X-irradiation in the retina of the rat.

In this study Lister rats were given doses of X-rays ranging from 200-2,000 Rads to the retina of one eye, sacrificed at various time intervals between one hour and one month later and the irradiated eye processed for electron microscopy. The rod photoreceptor cells were by far the most radiosensitive cells in the retina, their outer segments showing distinctive membrane damage at one hour after 200 Rads of X-rays. Photoreceptor cell death was not seen at doses less than 1,000 Rads in the time period of the experiment. The retinal pigment epithelial (RPE) cells showed damage in the form of mitochondrial swelling but only in doses over 500 Rads. Retinal pigment epithelial cell loss did not occur under 2,000 Rads. The inner retinal neurones, glial elements and the retinal vasculature did not show any ill effects in the time period of this study.

マウスにみられる自然発症型変形性膝関節症 ＩＩ 電子顕微鏡による細胞の変化の検討

Articular cartilage and subchondral bone of spontaneous osteoarthritis of ICR mouse knees were studied by transmission electron microscopy. The most characteristic finding of this mouse knee was chondrocyte necrosis of calcified cartilage before apparent osteoarthritic changes of cartilage. Early ultrastructural changes of the osteoarthritic cartilage included degeneration and disappearance of superficial discoid chondrocytes, microscar formation in the superficial layer, degeneration of deep-layer chondrocytes, and atrophy or degeneration of osteocytes in the subchondral bone. In advanced osteoarthritis, there was a various degree of cartilage loss and most of the cells in remaining cartilage and subchondral bone were necrotic. These findings resembled aging changes of the articular cartilage, however, the changes seen in osteoarthritic cartilage were thought to be severer, more frequent and more extensive than those in aging cartilage. Though proliferative changes such as chondrocyte clusters and osteophyte formation were not observed, there were some chondrocytes with abundant rough endoplasmic reticulum, mitochondria and Golgi complex. Densely packed thick collagen fibrils were often seen in contact with those cells. These thick collagen fibrils, which are similar to fibrils found in microscars, may be formed as a result of reparative reaction of cartilage chondrocytes.