Zonal heterogeneity of calcium distribution in rat hepatocytes: an electron microscopic study with a combined glutaraldehyde-osmium-pyroantimonate technique.

Potassium pyroantimonate in combination with osmium tetroxide is an excellent technique for investigation of the subcellular distribution of calcium ions in glutaraldehyde-fixed liver tissue. Under appropriate incubation conditions potassium ions are replaced by calcium ions to form an insoluble electron-dense calcium antimonate precipitate. The presence of calcium within this antimonate precipitate is confirmed by the electron spectroscopic imaging (ESI) technique. Chelator treatment with EGTA [ethylene glycol-bis-(beta-amino-ethyl ether) N',N',N'-tetraacetic acid] is usually used as a control, preventing the precipitation of calcium. In our investigation, computerized image analysis revealed a gradient of the calcium distribution from the periportal to the pericentral area. A finely dispersed precipitate was found in the mitochondrial matrix and in the euchromatin of the nuclei of the periportal hepatocytes, and in the organelles of the pericentral area a coarser deposit in lower concentration was recognized. Our findings indicate that periportal and pericentral hepatocytes retain their zonal characteristics also with respect to calcium concentration in mitochondria and nuclei.

Sephadex induced bronchial hyperreactivity in the rat: hematology, histology, histochemistry and immunohistology of the lung.

24 hours after an i.v. injection of 2 mg Sephadex G 200 particles ovalbumin sensitized Sprague Dawley rats show an antigen specific bronchial hyperreactivity and an unspecific hyperreactivity against serotonin. The aim of this study was to investigate the effects of Sephadex on blood parameters and lung pathology to find the morphological substrate of bronchial hyperreactivity in this animal model. In the blood neutrophilia (p < 0.01) but no eosinophilia was present. We conclude that a blood eosinophilia needs not to be necessarily correlated with hyperreactivity of the airways like claimed by other investigators for this animal model. Histologically we found that Sephadex particles are trapped in smaller-diameter arteries of the lung and lead to a granulomatous arteritis consisting mainly of ED1 positive and widely ED2 negative macrophages interspersed with eosinophils and neutrophils. Larger vessels not occluded by particles showed perivascular oedema with infiltration of eosinophils. We report here for the first time a significant hypertrophy of PAS positive goblet cells (p < 0.01) accompanied by a peribronchial infiltration with eosinophils (p < 0.01) and macrophages positive for ED1, ED2 and Ox-6 (p < 0.01) but not Ox-19 positive T-lymphocytes. The authors suggest that the peribronchial inflammation contributes importantly to the onset of bronchial hyperreactivity in this animal model and that the hypertrophy of goblet cells indicates the pathophysiological importance of peribronchial leukocytes.

[Aplastic anemia in ferrets]. / Ein Beitrag zur aplastischen Anämie beim Frettchen.

The present report describes both estrus associated and estrogen induced aplastic anemia in 7 female ferrets. The animals were presented with anemia, anorexia, apathy, melena and subcutaneous hemorrhages. Histologically, the animals showed panmyelophthisis and cystic endometrial hyperplasia.

[Megakaryocytic leukemia in three cats]. / Megakaryozytenleukose bei drei Katzen.

The present report describes a myeloproliferative disease, type megakaryocytic leukemia in three cats, 4, 8 and 11 years old. Clinically, the animals showed severe anemia. Neoplastic infiltrates were present in bone marrow, spleen, lymphnodes, liver and kidney. Two animals were FeLV positive.

[Epibulbar melanoma in a cat]. / Epibulbäres Melanom bei einer Katze.

In a nine-year-old female cat, the diagnostic procedure, therapy and differential diagnoses of a rare, primary, malignant, epibulbar melanoma are presented. Post-operatively no recidivation or metastases occurred within 18 months.

The effect of verapamil on mitochondrial calcium content in normoxic, hypoxic and reoxygenated rat liver.

Calcium channel blockers protect cells against ischaemia-reperfusion injury. In the present study, the effect of verapamil on mitochondrial calcium content was investigated in situ in normoxic, hypoxic and reoxygenated rat liver. Subcellular distribution of exchangeable calcium ions, which form an electron-dense precipitate with antimonate, was demonstrated with the glutaraldehyde-osmium antimonate technique. Calcium precipitates were quantified morphometrically using automatic image analysis. In normoxic liver, the mitochondrial calcium content formed a gradient decreasing from the periportal to perivenous regions. The low mitochondrial calcium content in perivenous regions remained unaffected in all experimental conditions. In hypoxic and reoxygenated liver, the calcium content in mitochondria of the periportal areas was significantly reduced. Verapamil pretreatment levelled the calcium gradient in normoxic liver by reducing the periportal calcium content. Verapamil had no effect on the mitochondrial calcium content in hypoxic liver. In contrast, in verapamil-pretreated reoxygenated liver, the mitochondrial calcium content in periportal mitochondria increased significantly, thus restoring the zonal calcium gradient. In conclusion, these data suggest that modulations of mitochondrial calcium content in the periportal region of the liver lobule may play an important role in the protective effects of verapamil against ischaemia-reperfusion injury.