ULTRASTRUCTURAL TRANSFORMATION IN MITOCHONDRIA ISOLATED FROM KIDNEYS OF NORMAL AND LEAD-INTOXICATED RATS

Mitochondria isolated from kidneys of lead-intoxicated rats have been shown to have decreased oxidative and phosphorylative abilities. The purpose of this study was to determine whether these abnormal mitochondria would undergo ultrastructural transformation during controlled respiration in the absence of phosphate acceptor (State IV), as previously demonstrated for normal liver mitochondria. It was first shown that normal rat kidney mitochondria transforms from a condensed ultrastructural conformation to an orthodox conformation after 5 min of State IV respiration with pyruvate-malate substrate. Reversal to a condensed conformation follows stimulation of respiration with adenosine diphosphate (ADP). A large portion of kidney mitochondria from lead-poisoned rats do not change from condensed to orthodox conformation during State IV respiration. Other mitochondria do transform to the orthodox form but they rapidly degenerate. State IV respiration decreases as these few orthodox mitochondria disintegrate. The conclusion is that those mitochondria that do not undergo change in ultrastructure have impairment of electron transport, and that those that do become orthodox have increased membrane lability and undergo degeneration.     

THE STRUCTURE OF EOSINOPHIL LEUKOCYTE GRANULES IN RODENTS AND IN MAN

The structure of the specific granules of eosinophil leukocytes has been studied by electron microscopy in sections of tissues, buffy coats, and sediments of peritoneal washings of rats, mice, guinea pigs, and men. The core of eosinophil granules is a crystal which has a cubic lattice with a repeat of ~30 A in rodents and ~40 A in man. The chemical composition of the core is discussed in connection with recent cell fractionation studies, and the hypothesis that the core is a crystal of peroxidase is considered.     

THE DEGENERATION AND REAPPEARANCE OF MITOCHONDRIA IN THE EGG CELLS OF A PLANT

Details are given of the elimination of mitochondria which occurs during the first 1 to 2 hours of the life of the egg of the fern Pteridium aquilinum (L.) Kuhn. During this phase the only mitochondria present are swollen and appear degenerate, but subsequently the cytoplasm of the egg becomes filled with large mitochondria containing numerous villi. Accompanying the appearance of these mitochondria, many, if not all of which have a peculiar umbo-like form, is the production by the nucleus of conspicuous and complex evaginations. The umbo-mitochondria are believed to be new, and a mechanism is suggested by which they may be generated from the complex evaginations of the nucleus.     

VARIATIONS IN THE STRUCTURE OF MITOCHONDRIA

A characteristic internal structure, consisting of a double-layered outer wall enclosing a matrix-filled space through which pass double-layered membranous folds, would appear to comprise as satisfactory a definition of mitochondria for electron microscopy as their intravital affinity for Janus green affords for light microscopy. Relying for identification upon this characteristic internal structure, mitochondria appear to be pleomorphic structures which vary in size, shape, complexity, and density. They are labile also in that their number may increase or decrease under controlled conditions. The possibility therefore exists that these organelles are constantly being formed and destroyed, perhaps by their participation in metabolic processes. The problem of the origin of mitochondria is in an unsatisfactory state. New organelles unquestionably are formed in particular physiological states. The possibility that new bodies are produced by fission of ones already present does not seem adequate. On the other hand, the possible fabrication of new mitochondria out of intracellular membranes, although an attractive hypothesis, has not been adequately substantiated.     

THE USE OF FORMALIN FIXATION IN THE CYTOCHEMICAL DEMONSTRATION OF SUCCINIC AND DPN- AND TPN-DEPENDENT DEHYDROGENASES IN MITOCHONDRIA

Brief formalin fixation in the cold prior to histochemical assay of rat liver and pancreas for various dehydrogenases has been used successfully to circumvent the structural damage and enzymatic loss to which mitochondria of frozen sections would otherwise be subject. To obtain an optimal result a single set of conditions has been devised, including fixation prior to freezing of minute (finely diced) organ blocks in graded concentrations (0.7 to 2.0 per cent) of formaldehyde in chilled (1–4°C) Hanks'' balanced salt solution, freezing at not higher than -70°C, and use of nitro-BT or, preferably, tetranitro-BT. The present histochemical study of hepatic and acinar cells indicates that not only are succinic and D-β-hydroxybutyric dehydrogenases located exclusively in the mitochondria but so are lactic, malic, and the isocitric dehydrogenases.     

THE STRUCTURE AND FORMATION OF PROTEIN GRANULES IN THE FAT BODY OF AN INSECT

In the larva of the butterfly Calpodes ethlius, the fat body begins to store protein in the form of granules at about 30 to 35 hours before pupation, at a time when the endocuticle is being resorbed. At least two sorts of granule can be distinguished. The first granules to arise are those within vesicles of the Golgi complex. These may increase in size by incorporating material from microvesicles at their surface and by coalescence with one another. Later, at about 10 hours before pupation, another sort of granule arises by the isolation of regions of the endoplasmic reticulum (ER) within paired membranes derived from Golgi vesicles. Several of these ER isolation bodies coalesce, with fusion of their outer isolating membranes. The ribosomes and membranes may then disappear and the granules become indistinguishable from the protein granules formed from Golgi vesicles, or the ribosomes may remain and be embedded in dense crystalline protein, forming a storage body for both protein and RNA. Mitochondria are isolated within paired membranes in the same way as regions of the ER. The isolated mitochondria also coalesce in a similar manner. When the inner membranes are lost, the structure of a group of isolation bodies is indistinguishable from that of a cytolysome. Isolation within paired membranes, as described here, may be of general importance in segregating regions of massive lysis or massive sequestration.     

钙调素参与玉米线粒体琥珀酸脱氢酶活性的调节

经DEAE C-32柱纯化的玉米(Zea mays L.)线粒体琥珀酸脱氢酶(SDH),用NAD激酶(NADK)法测定时,无钙调素(CaM)活性,说明不存在游离CaM;而用ELISA法测定总CaM时,却可检测到CaM。纯化的SDH加热处理后,能激活NADK,可能加热释放出游离CaM。纯化SDH的电泳分析表明,天然聚丙烯酰胺凝胶电泳(PAGE)只显示1条主带;而SDS-PAGE则出现67.0kD、30.0kD、16.7kD 3条带,前两条带与SDH的大、小亚基分子量一致,第三条带与CaM电泳迁移率一致。上述结果说明CaM可能与SDH处于结合状态,而且其活性受CaM调节。     

THE EFFECTS OF ERYTHROMYCIN AND CHLORAMPHENICOL ON THE ULTRASTRUCTURE OF MITOCHONDRIA IN SENSITIVE AND RESISTANT STRAINS OF PARAMECIUM

The effects on cell structure of 12 hr to 6 days of exposure to erythromycin or chloramphenicol, two antibiotics known to inhibit specifically the mitochondrial protein synthesizing system, have been studied in the ciliate Paramecium aurelia. A wild type strain (sensitive to both antibiotics) and three mutant strains carrying cytoplasmically inherited mutations conferring resistance to one or the other antibiotic have been used In sensitive cells both antibiotics lead to a progressive and profound alteration of mitochondrial structure evidenced by an elongation of the organelle, a considerable decrease in the number of cristae, and the appearance of some abnormal lamellar cristae and of rigid plates of periodic structure. The modifications of cell structure, then, are mainly restricted to mitochondrial cristae. The three resistant mutants studied, on the contrary, retain normal or nearly normal mitochondrial structure in the presence of the antibiotic to which they are resistant. This fact is in good agreement with the postulated location in the mitochondrial DNA of the resistance mutations studied. The results are discussed in the light of present knowledge concerning the function of the mitochondrial protein-synthesizing system.     

ELECTRON-OPAQUE FIBRILS AND GRANULES IN AND BETWEEN THE CELL WALLS OF HIGHER PLANTS

The components of higher-plant cell walls which become electron-opaque after staining with ruthenium-osmium were studied by electron microscopy. A fibrillar material which absorbs this stain is a major wall constituent in the root epidermal cells of carrot and morning glory. In both form and size, these fibrils resemble those found on the surface of suspension-cultured cells of the same species Some cells of woody species show an irregular distribution of electron-opaque material in the cell wall matrix and middle lamella. This material, which has an amorphous appearance with many electron stains, is shown by ruthenium-osmium staining to be an aggregate of discrete granules, 150–220 A in diameter. These observations are not consistent with the concept of the cell wall matrix and middle lamella as an amorphous, uniform gel     

整合素相关激酶在糖尿病肾病的表达及其意义

目的探讨整合素相关激酶(Integrin-Linked Kinase,ILK)在糖尿病肾病患者肾组织中的表达及其意义.方法对3例正常肾组织,14例糖尿病肾病患者肾穿刺活检标本,应用免疫组织化学方法检测ILK和FN在肾组织的阳性表达强度,并作图像分析处理.结果在正常肾组织,ILK主要表达于肾小球脏层上皮细胞,系膜细胞和小管上皮细胞呈弱表达.在糖尿病肾病,ILK表达于肾小球脏层上皮细胞和系膜细胞,在萎缩变性的肾小管上皮细胞表达增强.在肾小球结节硬化时,ILK表达明显减少.此外,ILK和FN的表达量在糖尿病肾病早、中期成正相关(P<0.001),在糖尿病肾病晚期成负相关(P<0.05).结论 ILK在糖尿病肾病肾组织中表达量显著增加,并与FN的表达有一定的相关性,说明其可能通过促进细胞外基质FN等的积聚,在糖尿病肾小球硬化过程中发挥重要作用.     

THE DEVELOPMENT OF PIGMENT GRANULES IN THE EYES OF WILD TYPE AND MUTANT DROSOPHILA MELANOGASTER

The eye pigment system in Drosophila melanogaster has been studied with the electron microscope. Details in the development of pigment granules in wild type flies and in three eye color mutants are described. Four different types of pigment granules have been found. Type I granules, which carry ommochrome pigment and occur in both primary and secondary pigment cells of ommatidia, are believed to develop as vesicular secretions by way of the Golgi apparatus. The formation of Type II granules, which are restricted to the secondary pigment cells and contain drosopterin pigments, involves accumulation of 60- to 80-A fibers producing an elliptical granule. Type III granules appear to be empty vesicles, except for small marginal areas of dense material; they are thought to be abnormal entities containing ommochrome pigment. Type IV granules are characteristic of colorless mutants regardless of genotype, and during the course of development they often contain glycogen, ribosomes, and show acid phosphatase activity; for these reasons and because of their bizarre and variable morphology, they are considered to be autophagic vacuoles. The 300-A particles commonly found in pigment cells are identified as glycogen on the basis of their morphology and their sensitivity to salivary digestion.     

THE DISTRIBUTION OF HISTOCHEMICALLY DEMONSTRABLE SUCCINIC DEHYDROGENASE AND OF MITOCHONDRIA IN TONGUE AND SKELETAL MUSCLES

Various skeletal muscles show considerable variations in histochemically demonstrable succinic dehydrogenase activity. In muscles from the lower extremities, the back, and the abdominal wall, only some fibers reveal evidence of enzymatic staining. In the diaphragm and masseter more fibers react positively. In the tongue, however, all fibers show marked activity comparable to that found regularly in the heart. There is a close similarity in the distribution of histochemically demonstrable succinic dehydrogenase and stainable mitochondria in tissue sections.