Environmentally induced changes in mitochondria and endoplasmic reticulum of Saccharomyces carlsbergensis yeast

The effects of culture environment on the volume density and surface density of mitochondria and endoplasmic reticulum in a facultative yeast were studied. When compared with cells grown aerobically on a nonrepressive substrate, cells grown in the absence of oxygen showed a sharp reduction in both volume density of mitochondria and surface density of the inner mitochondrial membrane (imm) in the remaining mitochondrial profiles. Use of fermentable (repressive) substrates under aerobic conditions restricted the volume density of mitochondria to a much greater extent than the surface density of imm. The range of mitochondrial volume densities in these experiments was 4-11%. Surface density of endoplasmic reticulum (ER) was sensitive to growth rate and in particular to changes in oxygen tension, showing large fluctuations during both anaerobic and aerobic adaptation. These fluctuations in ER are discussed in relation to the known role of this organelle in lipid metabolism.     

蚕豆萎焉病毒2感染豌豆叶细胞后引起的线粒体增生和聚集

应用超薄切片和免疫金标记电镜技术,结合体视学分析研究了受蚕豆萎焉病毒2(BBWV 2) 中国分离物B935侵染的豌豆(Pisum sativum)叶细胞中线粒体的异常变化。结果表明,感病细胞线粒体增生并聚集于细胞质的膜增生区周围,体积增大,形状畸变,一些线粒体内含有类结晶包涵体。病叶细胞与健康对照之间线粒体的体积密度(VV)、表面积密度(SV)、数密度(NV)等参数存在显著差异(P<0.01),而形状因子(PE)、周长指数(CI)、比表面积(RSV)等参数随不同病变阶段而有变化。在线粒体周围及线粒体之间的网格结构可被BBWV 2金标记抗体特异性标记．推断为正在组装的病毒粒子。子代病毒形成结晶体和管状体,有高密度的免疫金颗粒标记。上述研究结果提示BBWV 2 引起的细胞线粒体异常变化与病毒复制组装有关,聚集线粒体的外膜粘连面可能是病毒粒子组装部位,一些线粒体内的类结晶包涵体可能代表了某种蛋白质异常积累。     

蚕豆萎焉病毒2感染豌豆叶细胞后引起的线粒体增生和聚集

应用超薄切片和免疫金标记电镜技术,结合体视学分析研究了受蚕豆萎焉病毒2（BBWV2）中国分离物B935侵染的豌豆（Pisumsativum）叶细胞中线粒体的异常变化。结果表明,感病细胞线粒体增生并聚集于细胞质的膜增生区周围。体积增大,形状畸变,一些线粒体内含有类结晶包涵体。病叶细胞与健康对照之间线粒体的体积密度（Vv）、表面积密度（Sv）、数密度（Nv）等参数存在显著差异（P〈0．01）,而形状因子（PE）、周长指数（CI）、比表面积（Rsv）等参数随不同病变阶段而有变化。在线粒体周围及线粒体之间的网格结构可被BBWV2金标记抗体特异性标记。推断为正在组装的病毒粒子。子代病毒形成结晶体和管状体,有高密度的免疫金颗粒标记。上述研究结果提示BBWV2引起的细胞线粒体异常变化与病毒复制组装有关。聚集线粒体的外膜粘连面可能是病毒粒子组装部位。一些线粒体内的类结晶包涵体可能代表了某种蛋白质异常积累。     

Effect of cold environment on skeletal muscle mitochondria in growing rats

Summary Growing rats (4 weeks old) were kept for 3 weeks at 11° C and 24° C respectively. The cold-adapted animals showed a significantly higher oxygen consumption (64%). Volume density of subsarcolemmal and interfibrillar mitochondria as well as volume density of fat droplets were estimated in M. soleus and the diaphragm of both groups. In cold-adapted animals, the total volume of mitochondria was significantly increased by 24% in diaphragm and 37% in M. soleus. The volume of subsarcolemmal mitochondria was almost doubled in each muscle, but the volume of interfibrillar mitochondria did not change significantly. The surface of the inner mitochondrial membranes per unit volume of mitochondrion in M. soleus was significantly increased both in interfibrillar and subsarcolemmal mitochondria, whereas the surface of the outer mitochondrial membranes per unit volume of mitochondrion was increased only in the subsarcolemmal mitochondria. The volume of fat droplets in the diaphragm and M. soleus of cold adapted animals increased significantly by 62% and 150% respectively.     

Structure and function of mitochondria in hepatopancreas cells from metabolically depressed snails

Mitochondria in cells isolated from the hepatopancreas of aestivating land snails (Helix aspersa) consume oxygen at 30% of the active control rate. The aim of this study was to investigate whether the lower respiration rate is caused by a decrease in the density of mitochondria or by intrinsic changes in the mitochondria. Mitochondria occupied 2% of cellular volume, and the mitochondrial inner membrane surface density was 17 microm(-1), in cells from active snails. These values were not different in cells from aestivating snails. The mitochondrial protein and mitochondrial phospholipid contents of cells were also similar. There was little difference in the phospholipid fatty acyl composition of mitochondria isolated from metabolically depressed or active snails, except for arachidonic acid, which was 18% higher in mitochondria from aestivating snails. However, the activities of citrate synthase and cytochrome c oxidase in mitochondria isolated from aestivating snails were 68% and 63% of control, respectively. Thus the lower mitochondrial respiration rate in hepatopancreas cells from aestivating snails was not caused by differences in mitochondrial volume or surface density but was associated with intrinsic changes in the mitochondria.     

Structural changes in L-cell mitochondria exposed to cyanide

Using light and electron microscopy, as well as stereological analysis, a study was made of structural changes in mitochondria of cultured L-cells acted upon with 1 mM cyanide. After 23 hours of incubation, in cells cultured in the presence of cyanide the volume density of mitochondria, the volume of the average mitochondrion and the surface area of its membranes increased nearly twice as much. Concurrently, the number of mitochondria decrease also practically two-fold without any reliable changes in the surface area of membranes per unit of mitochondrion volume. A shorter (7 hours) incubation of L-cells in the presence of cyanide results in a decreased volume density of mitochondria in the cytoplasm, decreased volume of the average mitochondrion, without any significant changes in other above mentioned parameters. It is supposed that the changes in mitochondria under a prolonged cyanide treatment of cells are conditioned by the fusion of mitochondria as well as by a compensatory increase in the total surface of mitochondrial membranes.     

Morphological differentiation of mitochondria in the early chick embryo: a stereological analysis

The morphological evolution of mitochondria in three cell types of chick embryo in neurulation was analyzed by stereological methods. Mitochondria, showing a random distribution, were characterized by moderate electron-dense matrices and normal cristae. The numerical density of mitochondria significantly increased in the neuroectoderm and epiblastic cells while their volume density remained unchanged. The mitochondria in mesoderm cells were ellipsoidal (axial ratio 2:1) at stages 5 and 8 although they underwent an elongation in neuroectoderm and epiblastic cells (axial ratio from 2:1 to 1.6:1). The individual size of "average mitochondria" in the mesoderm cells was smaller than in other cell types. The total V/S (volume/surface) ratio of mitochondria decreased during neurulation. These morphological changes have been discussed emphasizing the possible metabolical role of mitochondria during morphogenesis.     

BALB/C小鼠肝细胞结构的年龄变化——光镜与电镜的计量形态学研究

作者研究了1月、6月和12月三个不同年龄组的BALB/c小鼠的肝细胞结构。利用计量形态学方法在光镜与电镜两个水平上对肝细胞整体及重要细胞器的年龄变化进行定量分圻,发现在所观察的结构参数中大部分随年龄而发生动力学变化。文中讨论了某些变化的可能意义。     

Autophagy of mitochondria in rat bone marrow erythroid cells Relation to nuclear extrusion

Summary Late erythroblasts and reticulocytes from bone marrow of male Wistar rats were studied by electron-microscopic stereology. Late erythroblasts with morphological signs of nuclear extrusion (EN+erythroblasts) and late erythroblasts without these signs (EN-erythroblasts) were analysed separately. The volumes of mitochondria, autophagosomes, autophagocytosed mitochondria, autophagocytosed cytoplasm and degraded material inside autophagosomes were calculated per unit volume of cytoplasm.The results demonstrate that (1) the volume density of mitochondria in the cytoplasm decreases by 34% during maturation from (EN-)- to (EN+)-erythroblasts (p< 0.001) and by 60% during differentiation from (EN+)-erythroblasts to reticulocytes (p<0.001), (2) a fivefold increase in the volume density of autophagosomes in the cytoplasm is noted during maturation from (EN-)- to (EN+)-erythroblasts (p<0.01), whereas the value of this parameter remains essentially unchanged during the subsequent differentiation to reticulocytes, (3) no mitochondria are found inside autophagosomes of (EN-)-erythroblasts, whereas mitochondria occupy 26% and 35%, respectively, of the autophagosomal volume in (EN+)-erythroblasts and in reticulocytes.Our results show that autophagocytosis of mitochondria starts at the moment of nuclear extrusion and continues in the bone marrow reticulocytes.     

Stereological studies on germ cell mitochondria of rat during spermatogenesis and spermiogenesis

The following stereological parameters of mitochondria were calculated in rat germ cells during the spermatogenesis: volume density of matrix, outer compartment, outer membrane and inner membrane, surface density of outer membrane and inner membrane. They were the basis to calculate the partition coefficient of matrix and partition coefficient of outer compartment. The matrix volume demonstrated a decreasing in mitochondria of germ cells during spermatogenesis. The relative volume of outer compartment was calculated with the intracristal spaces and revealed increasing from spermatogonia to spermatids. The partition coefficient for the matrix significantly decreased. Our observations suggest that transformation of mitochondrial configuration during spermatogenesis and spermiogenesis is the expression of intensive metabolic processes and activity of membrane transport in germ cells.     

Physiological Cost of Physical Exercise and Mitochondrial Volume in Working Muscles of Subjects Exposed to Long-Term Hypokinesia: Effects of Local Resistance Exercise

The results of changes in the physiological cost of 30-min submaximal aerobic bicycle ergometric exercise and characteristics of the mitochondrial apparatus of m. vastus lateralis were assessed comparatively during 120-day (–6°) antiorthostatic hypokinesia either without prophylactic measures or with low-intensity resistance exercise training for 60 days using a Penguin exercise suit. Hypokinesia was accompanied by an increase in the working heart rate and lactate accumulation in the blood during the test exercise, as well as by a decrease in the myofibril size and the volume density of mitochondria in the m. vastus lateralis fibers. The patterns of dynamic changes in the lactate concentration in the blood during exercise training and in the volume density of central mitochondria were found to be similar. A correlation between the rate of lactate accumulation in the blood during the test exercise and the volume density of mitochondria in the working muscle appeared after long-term (60 days) exposure to hypokinesia. The use of the Penguin exercise suit in dynamic mode during prolonged (60-day) exposure to hypokinesia completely prevented the following effects: atrophy of slow-type fibers, a decrease in the volume density of central mitochondria, and an increase in the level of lactate accumulation in the blood under conditions of a standard submaximal aerobic exercise load. The correlation links between the oxidative potential of working muscle and the energy supply of muscular work are discussed.     

Quantitative immunocytochemical studies on differential induction of serine:pyruvate aminotransferase in mitochondria and peroxisomes of rat liver cells by administration of glucagon or di-(2-ethylhexyl)phthalate

Differential induction of serine: pyruvate amino-transferase (SPT) in rat liver parenchymal cells by administration of glucagon or di-(2-ethylhexyl)phthalate (DEHP) was studied using post-embedding immunocytochemical techniques and morphometric methods. Two groups of rats were fasted for 5 days and daily received peritoneal injection of glucagon (300 micrograms/100 g) or physiological saline. Another two groups of rats were fed on laboratory chow with or without 2% DEHP for 2 weeks. Livers were perfusion-fixed, cut into tissue sections (50-100 micron), and processed to cytochemistry for catalase, immunocytochemistry for SPT, and conventional procedures for electron microscopy. The morphometric analysis showed that glucagon injection has negligible effect on the volume and numerical density and mean diameter of peroxisomes, whereas volume density of mitochondria was decreased by 25%. By DEHP administration peroxisomes were about 3-fold increased in the volume and numerical density. Mitochondria was increased about 40% in the numerical density, but unchanged in the volume density. Light and electron microscopic immunocytochemistry demonstrated that glucagon injection exclusively enhanced mitochondrial SPT, whereas DEHP administration exclusively induced in peroxisomal SPT. Quantitative analysis showed that by the glucagon injection, the labeling density of mitochondria was increased about 4-fold, but that of peroxisomes was 1.6 times as much as control, while by DEHP administration, the labeling density of peroxisomes was enhanced about 3-fold but that of mitochondria was decreased by 13%. The results clearly indicate that glucagon induces mitochondrial SPT, whereas peroxisome proliferator, DEHP induces peroxisomal SPT.     

CYTOLOGICAL DIFFERENCES BETWEEN A CYTOPLASMIC MALE STERILE TOBACCO CYBRID AND ITS FERTILE COUNTERPART DURING EARLY ANTHER DEVELOPMENT

Ultrastructural differences were detected between a cytoplasmic male sterile tobacco cybrid (Nicotiana sp.) formed by protoplast fusion and normal, fertile tobacco. Cell structure was compared between anther primordia from normal, fertile tobacco and anther primordia from the cybrid using stereological methods. Particular emphasis was placed on the ultrastructure of mitochondria because of their known relationship to cytoplasmic male sterility in this cybrid and other plants. The volume density of mitochondria in cybrid anther primordia (6.3%) was significantly lower than in normal, fertile anther primordia (10.1%), although mitochondria from both plants contained similar amounts of cristae and profiles were of similar relative area. Dictyosomes and vacuoles also differed in volume density but not at a statistically significant level. Although the volume density of plastids did not differ, a larger amount of starch was stored within plastids in cybrid anther primordia than in normal, fertile anther primordia. These results are compatible with the hypothesis that an abnormally low rate of mitchondrial replication, and the resultant limit on adenosine triphosphate production, could contribute to cytoplasmic male sterility in the cybrid.     

Lung oxygen consumption and mitochondria of alveolar epithelial and endothelial cells.

We examined oxygen consumption by lung slices and measured the volume density of mitochondria of granular pneumocytes, alveolar type I cells, and alveolar capillary endothelial cells in several species. We found that lung oxygen consumption (mu-1 02 times h-1 times mg DNA-1) varies inversely with the log of animal body weight and with the species alveolar diameter and directly with the species respiratory rate. The volume density of granular pneumocyte mitochondria show a direct linear correlation with the lung's oxygen consumption and the species respiratory rate, and an inverse linear correlation with the species alveolar diameter. The volume density of mitochondria in type I alveolar epithelial cells and capillary endothelial cells, considered together, did not differ in the two species studied (mouse and rat). We conclude that there are interspecies differences in oxygen consumption by lung cells and that granular pneumocytes contribute to these differences. We suggest that, at least part of these differences, are related to interspecies differences in surfactant secretory activity.     

Volume density and distribution of mitochondria in harbor seal (<Emphasis Type="Italic">Phoca vitulina</Emphasis>) skeletal muscle

Recent studies have shown that harbor seals (Phoca vitulina) have an increased skeletal muscle mitochondrial volume density that may be an adaptation for maintaining aerobic metabolism during diving. However, these studies were based on single samples taken from locomotory muscles. In this study, we took multiple samples from a transverse section of the epaxial (primary locomotory) muscles and single samples from the m. pectoralis (secondary locomotory) muscle of five wild harbor seals. Average mitochondrial volume density of the epaxial muscles was 5.6%, which was 36.6% higher than predicted for a terrestrial mammal of similar mass, and most (82.1%) of the mitochondria were interfibrillar, unlike athletic terrestrial mammals. In the epaxial muscles, the total mitochondrial volume density was significantly greater in samples collected from the deep (6.0%) compared with superficial (5.0%) regions. Volume density of mitochondria in the pectoralis muscle was similar (5.2%) to that of the epaxial muscles. Taken together, these adaptations reduce the intracellular distance between mitochondria and oxymyoglobin and increase the mitochondrial diffusion surface area. This, in combination with elevated myoglobin concentrations, potentially increases the rate of oxygen diffusion into mitochondria and prevents diffusion limitation so that aerobic metabolism can be maintained under low oxygen partial pressure that develops during diving.     

Cytological effects of platelet-derived growth factor on mitochondrial ultrastructure in fibroblasts

The goal of this study was to evaluate morphofunctional changes in mitochondrial ultrastructure after platelet-derived growth factor application in fibroblasts as an indicator of mitochondrial activation in processes like wound healing. NRK-49F fibroblasts were synchronized, incubated with PDGF (platelet-derived growth factor) and studied by electron microscopy. Volume density (Vv), numerical density (Nv) and surface density (Sv) were measured by stereological analysis. Application of PDGF on NRK-49F caused an increase in mitochondrial volume density by 57% and surface area of cristae per mitochondrion by 65%. The numerical density of the mitochondria was decreased in the PDGF-treated cells by 23%, but at the same time their mean volume was increased. Furthermore, the mitochondria had a complex and highly variable shape both in control and PDGF-treated cells, possibly indicating the existence of a mitochondrial reticulum. The results demonstrated that biochemically active membrane systems in fibroblast mitochondria are enlarged as a direct effect of small doses of platelet-derived growth factor and support the concept that this factor and related peptides serve as mitogens for connective tissue forming cells. Thus, in mitogenic processes like wound healing, the high energy demand of fibroblasts is provided by the increase of the inner surface of mitochondria.     

大鼠心肌线粒体缺血再灌损伤及SOD抗缺血再灌损伤作用的形态计量研究

研究了在体大鼠心肌缺血再灌模型心肌线粒体再灌损伤和SOD抗缺血再灌损伤的作用。实验分为:A组(缺血再灌组)B组(SOD组)和C组(假手术组)。电镜见A组线粒体高度肿胀、外膜缺损、嵴断裂溶解,出现无定形致密体、四膜嵴和杆状嵴等改变。B组线粒体轻度或中度肿胀、少数有轻度嵴溶解。个别线粒体内出现无定形致密体。三组线粒体超微结构立体计量数据比较,A组线粒体密度、线粒体体密度与肌原纤维体密度比率增高((P<0.01),线粒体比表面和比膜面降低(P<0.01)。自身动图像分析仪检测SDH反应灰度值A组呈强损伤反应,B组反应较轻,二者差别有显著性(P<0.05)。结果表明,心肌缺血再灌可致线粒体发生不可逆性损伤,SOD能减轻线粒体缺血再灌性损伤。     

Ultrastructure of neurons of the human sympathetic ganglia after sudden death (morphometric study)

Six cases of urgent autopsies after a sudden cardiac death (SCD) of persons at the age 40-60 years and 5 control cases have been investigated. Osmiophilic mitochondria with inclusions make 1% of the whole neuronal cytoplasm volume. If we take all mitochondria as 100%, volumetric density of mitochondria with osmiophilic inclusions makes 9.6%. Volumetric density of mitochondria at SCD in comparison with the control is significantly higher in connection with increase of their size. In other components of cytoplasm, in particular in lysosomes and lipofuscin, no changes are revealed.     

A comparative morphometric analysis of the effects of thyroxine and ACTH on the zona fasciculata of rat adrenal cortex.

The effects of thyroxine and ACTH on the ultrastructure of cells of the adrenal zona fasciculata were studied by morphometric analysis and compared by two-way analysis of variance. The results indicate that the effects of thyroxine and ACTH are different and independent. The statistically significant differences in effect of thyroxine and ACTH are: thyroxine causes an increase in the volume fraction of mitochondria and a decrease in the volume fraction of endoplasmic reticulum independent of ACTH; thyroxine causes a decrease in surface density of mitochondria and endoplasmic reticulum in the absence of ACTH, but in the presence of ACTH this effect is moderated by an interaction of throxine and ACTH. There were no statistically significant effects of either thyroxine or ACTH on volume fraction or surface density of liposomes.     

Stereological study of B-cell mitochondria in alloxan-treated mice.

Using stereological techniques, including semi-automatic image analysis, the B-cell mitochondria were studied in the pancreatic islets from one group of control mice and two groups of mice killed 10 min and 60 min, respectively, after alloxan administration. Ten min following alloxan the mitochondrial volume and envelope surface densities, the mean mitochondrial volume and surface area, and the area of mitochondrial profiles were significantly increased, whereas the mitochondrial numerical density was not significantly altered. At the 60 min observation time the mitochondrial volume density, the mean mitochondrial volume and surface areas, and the area of mitochondrial profiles were significantly decreased, whereas the mitochondrial envelope surface was not significantly altered. The findings indicate a rapid swelling, followed by disintegration of the mitochondria in the B-cells of alloxan-treated mice, thereby supporting our view that mitochondrial lesions play a primary role in the development of alloxan diabetes. These lesions are believed to be due to ionic alterations in the B-cells ("Pi-pH hypothesis").