Mitochondrial Surface Area Research Articles

Relative quantification of white blood cell mitochondrial DNA and assessment of mitochondria by use of transmission electron microscopy in English Springer Spaniels with and without retinal dysplasia

To compare relative amounts of WBC mitochondrial DNA (mtDNA; assessed via real-time PCR assay) and morphology of lymphocyte mitochondria (assessed via transmission electron microscopy [TEM]) in blood samples collected from English Springer Spaniels with and without retinal dysplasia. 7 and 5 client-owned English Springer Spaniels (1 to 11 years old) with and without retinal dysplasia, respectively. Blood samples were obtained from affected and unaffected dogs via venipuncture. Genomic DNA was extracted from WBCs of the 7 affected and 5 unaffected dogs, and relative quantification of the cytochrome c oxidase subunit 1 gene (COX1) was determined via analysis of real-time PCR results. White blood cells from 3 affected and 4 unaffected dogs were embedded in epoxide resin for TEM; cross sections were examined for lymphocytes, which were measured. The mitochondria within lymphocytes were quantified, and the mitochondrial surface area per lymphocyte cross section was calculated. A masked technique was used to compare mitochondrial morphology between the 2 groups. Compared with the smallest measured quantity of mtDNA among unaffected dogs, mtDNA amounts varied among unaffected (1.08- to 4.76-fold differences) and affected dogs (1- to 2.68-fold differences). Analysis of lymphocyte measurements and mitochondrial surface area, morphology, and quantity revealed no significant differences between affected and unaffected dogs. No significant differences were detected in relative amounts of WBC mtDNA or the size, number, or morphology of lymphocyte mitochondria in English Springer Spaniels affected with retinal dysplasia, compared with results for unaffected control dogs.

Stimulation of glutamate receptors in cultured hippocampal neurons causes Ca2+-dependent mitochondrial contraction

Cultured hippocampal neurons expressing mitochondrially-targeted enhanced yellow fluorescent protein (mito-eYFP) were used to quantitatively examine mitochondrial remodelling in response to excitotoxic glutamate. Mitochondrial morphology was evaluated using laser spinning-disk confocal microscopy followed by calibrated image processing and 3D image rendering. Glutamate triggered an increase in cytosolic Ca2+ and mitochondrial depolarization accompanied by Ca2+-dependent morphological transformation of neuronal mitochondria from “thread-like” to rounded structures. The quantitative analysis of the mitochondrial remodelling revealed that exposure to glutamate resulted in a decrease in mitochondrial volume and surface area concurrent with an increase in sphericity of the organelles. NIM811, an inhibitor of the mitochondrial permeability transition, attenuated the glutamate-induced sustained increase in cytosolic Ca2+ and suppressed mitochondrial remodelling in the majority of affected neurons, but it did not rescue mitochondrial membrane potential. Shortening, fragmentation, and formation of circular mitochondria with decreased volume and surface area accompanied mitochondrial depolarization with FCCP. However, FCCP-induced morphological alterations appeared to be distinctly different from mitochondrial remodelling caused by glutamate. Moreover, FCCP prevented glutamate-induced mitochondrial remodelling suggesting an important role of Ca2+ influx into mitochondria in the morphological alterations. Consistent with this, in saponin-permeabilized neurons, Ca2+ caused mitochondrial remodelling which could be prevented by Ru360.

Murine epidermal progenitor cells have a low redox state

The epidermis is a stratified cornified epithelium that protects the organism against physical and microbial insults. These environmental exposures can result in oxidative stress and affect cellular proliferation and differentiation. However, very few studies have evaluated the base line levels of reactive oxygen species (ROS) and antioxidant proteins in the epidermis or in the context of stem/progenitor cells, cellular types critical to the homeostasis of the epidermis. Here, we show that ROS are mainly produced by epidermal cells and that antioxidant proteins (MnSOD, GPx1, catalase) are differentially expressed in the epidermis. Using our previously sorted epidermal progenitor cells (side population, EpSPs) and more differentiated cells (non‐SPs), we show that catalase expression and activity are significantly decreased in EpSPs compared to non‐SPs. Furthermore, we found fewer peroxisomes per cell and decreased mitochondrial surface area in the EpSPs compared to non‐SPs. Using dihydroethidium (DHE), we show that EpSPs have less DHE signal compared to more differentiated cells. In summary, our data suggest that EpSPs have decreased levels of antioxidant proteins and ROS. Together, these studies indicate that the intracellular redox environment of EpSPs is reduced relative to their more differentiated progeny.Grant Funding SourceNational Institute of Health

Volume density and distribution of mitochondria in harbor seal (Phoca vitulina) 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.

Muscle fine structure may maintain the function of oxidative fibres in haemoglobinless Antarctic fishes.

Muscle fine structure and metabolism were examined in four species of Antarctic fishes that vary in their expression of haemoglobin (Hb). To determine how locomotory pectoral muscles maintain function, metabolic capacity, capillary supply and fibre ultrastructure were examined in two nototheniid species that express Hb (Notothenia coriiceps and Gobionotothen gibberifrons) and two species of channichthyid icefish that lack Hb (Chaenocephalus aceratus and Chionodraco rastrospinosus). Surprisingly, icefish have higher densities of mitochondria than red-blooded species (C. aceratus, 53+/-3% of cell volume; C. rastrospinosus, 39+/-3%; N. coriiceps, 29+/-3%; G. gibberifrons, 25+/-1%). Despite higher mitochondrial densities the aerobic metabolic capacities per g wet mass, estimated from measurements of maximal activities of key metabolic enzymes, are lower in icefish compared to red-blooded species. This apparent incongruity can be explained by the significantly lower mitochondrial cristae surface area per unit mitochondrion volume in icefishes (C. aceratus, 20.8+/-1.6 microm(-1); C. rastrospinosus, 25.5+/-1.8 microm(-1)) compared to red-blooded species (N. coriiceps, 33.6+/-3.0 microm(-1); G. gibberifrons, 37.7+/-3.6 microm(-1)). Consequently, the cristae surface area per unit muscle mass is conserved at approximately 9 m(2)g(-1). Although high mitochondrial densities in icefish muscle do not enhance aerobic metabolic capacity, they may facilitate intracellular oxygen movement because oxygen is more soluble in lipid, including the hydrocarbon core of intracellular membrane systems, than in aqueous cytoplasm. This may be particularly vital in icefish, which have larger oxidative muscle fibres compared to red-blooded nototheniods (C. aceratus, 2932+/-428 microm(2); C. rastrospinosus, 9352+/-318 microm(2); N. coriiceps, 1843+/-312 microm(2); G. gibberifrons, 2103+/-194 microm(2)). These large fibres contribute to a relatively low capillary density, which is partially compensated for in icefish by a high index of tortuosity in the capillary bed (C. aceratus=1.4, N. coriiceps=1.1).

Ultrastructural design of anuran muscles used for call production in relation to the thermal environment of a species

I examined the aerobic trunk muscles, which are used for call production, of male frogs from species that breed in different thermal environments to test the hypothesis that cold-adapted frogs should have fewer capillaries per unit mitochondrial volume in oxidative muscles than warm-adapted frogs because of reduced mitochondrial function at low temperatures. The species of interest were the cold-temperate Pseudacris crucifer and the warm-tropical Hyla microcephala in the family Hylidae, and the cold-temperate Rana sylvatica and the warm-temperate Rana clamitans in the family Ranidae. Trunk-muscle mitochondrial volume, V(V)(mt,f), was proportionally higher in species with higher mean calling rates (number of notes per hour), irrespective of the familial affinity of a species and the thermal environment in which it vocalized. Trunk-muscle capillary length density, J(V)(c,f), was significantly lower in P. crucifer than in H. microcephala because of significantly higher mean fiber area, a macro (f). Conversely, trunk-muscle J(V)(c,f) was similar in the two ranid species. Using total capillary length, J(c), and total mitochondrial volume, V(mt,m), as a measure of maximal oxygen supply and demand, respectively, in trunk muscles, J(c)-to-V(mt,m) ratios were significantly lower in cold-adapted P. crucifer (4.3 km cm(-3)) and R. sylvatica (4.8 km cm(-3)) than in warm-adapted H. microcephala (7.1 km cm(-3)) and R. clamitans (6.4 km cm(-3)). In contrast, J(c)-to-V(mt,m) ratios in the more anaerobic gastrocnemius muscle of these species was not related to the thermal environment of a species, which may reflect capillaries conforming to microcirculatory functions, e.g. lactate removal, that take precedence over oxygen delivery. Mitochondrial cristae surface area, S(V)(im,mt), in P. crucifer trunk and gastrocnemius muscles (37.7+/-1.6 and 35.9+/-1.5 m(2 )cm(-3) respectively) was, on average, similar to mammalian values, suggesting equivalent structural capacities of muscle mitochondria in these two taxa. Taken together, the present data suggest that trunk-muscle respiratory design may reflect a capillary supply commensurate with maximal levels of oxygen delivery set by mitochondria operating at different environmental temperatures. P. crucifer and H. microcephala trunk muscles were also characterized by a high lipid content, which contrasted with a near absence of trunk-muscle lipids in R. sylvatica and R. clamitans. The extraordinarily high lipid content of P. crucifer trunk muscles (26 % of muscle volume) may serve as an auxiliary oxygen pathway to mitochondria and thus compensate in part for this tissue's reduced capillary/fiber interface. The effect of potentially high depletion rates of trunk-muscle lipid stores on metabolic rates of male frogs while calling is discussed.

Effects of growth conditions on mitochondrial morphology in Saccharomyces cerevisiae.

Effects of growth conditions on mitochondrial morphology were studied in living Saccharomyces cerevisiae cells by vital staining with the fluorescent dye dimethyl-aminostyryl-methylpyridinium iodine (DASPMI), fluorescence microscopy, and confocal-scanning laser microscopy. Cells from respiratory, ethanol-grown batch cultures contained a large number of small mitochondria. Conversely, cells from glucose-grown batch cultures, in which metabolism was respiro-fermentative, contained small numbers of large, branched mitochondria. These changes did not significantly affect the fraction of the cellular volume occupied by the mitochondria. Similar differences in mitochondrial morphology were observed in glucose-limited chemostat cultures. In aerobic chemostat cultures, glucose metabolism was strictly respiratory and cells contained a large number of small mitochondria. Anaerobic, fermentative chemostat cultivation resulted in the large, branched mitochondrial structures also seen in glucose-grown batch cultures. Upon aeration of a previously anaerobic chemostat culture, the maximum respiratory capacity increased from 10 to 70 mumole.min-1.g dry weight-1 within 10 h. This transition resulted in drastic changes of mitochondrial number, morphology and, consequently, mitochondrial surface area. These changes continued for several hours after the respiratory capacity had reached its maximum. Cyanide-insensitive oxygen consumption contributed ca. 50% of the total respiratory capacity in anaerobic cultures, but was virtually absent in aerobic cultures. The response of aerobic cultures to oxygen deprivation was qualitatively the reverse of the response of anaerobic cultures to aeration. The results indicate that mitochondrial morphology in S. cerevisiae is closely linked to the metabolic activity of this yeast: conditions that result in repression of respiratory enzymes generally lead to the mitochondrial morphology observed in anaerobically grown, fermenting cells.

Inconsistent effectiveness of myocardial preservation among cardiac chambers during hypothermic cardioplegia

The purpose of this study was to examine the selective and differential natures of ischemic injuries among three cardiac chambers (right atrium, right ventricle, and left ventricle) from the viewpoint of ultrastructural morphometric study. Twenty consecutive adult patients undergoing cardiac operations were studied. The duration of aortic crossclamp time varied from 36 to 142 minutes (mean 83.4 +/- 36.4 minutes). Two serial specimens (preischemic and ischemic) were obtained from the right atrium, the right ventricle, and the left ventricle, respectively. A total of 120 biopsy specimens was obtained from these 20 patients. The average mitochondrial surface area of the left ventricle was 0.308 +/- 0.062 micron 2 in the preischemic stage and 0.352 +/- 0.083 microns 2 in the ischemic stage. This represented a 14.3% increase in mitochondrial surface area after ischemic injury (p less than 0.01). The mitochondrial surface area of the right ventricle showed an average increase of 43.7%, from 0.252 +/- 0.036 micron 2 in the preischemic stage to 0.362 +/- 0.087 micron 2 in the ischemic stage (p less than 0.0005). With respect to the mitochondrial surface area of the right atrium, there was an increase of 88.0%, from 0.217 +/- 0.044 micron 2 in the preischemic stage to 0.408 +/- 0.084 micron 2 (p less than 0.0005). The difference of mitochondrial swelling among three chambers was statistically significant (right atrium versus right ventricle versus left ventricle, p less than 0.0005). Moreover, the differences of mitochondrial swelling between any two chambers were also highly significant (right atrium versus right ventricle, p less than 0.0005; right ventricle versus left ventricle, p less than 0.01; right atrium versus left ventricle, p less than 0.0005). In conclusion, our findings suggest that from the viewpoint of ultrastructural morphometric study myocardial injury after an average of 83 minutes of ischemic arrest is poorer in the right chambers of the heart than in the left ventricle, with the right atrium having the poorest preservation.

Trypanosoma (Nannomonas) congolense: Changes in respiratory metabolism during the life cycle

All four life cycle stages (bloodstream, procyclic, epimastigote, and metacyclic) of Trypanosoma congolense IL 3000 were assayed with an oxygen electrode (polarograph) for the presence of terminal oxidases and carbon-source preference. In addition, these stages were used for histochemical analysis of mitochondrial activity using rhodamine 123, nitroblue tetrazolium, and diaminobenzidine. Morphometry was used to compare mitochondrial volumes and surface area among the different life cycle stages. It was found that in contrast to epimastigote forms, which were metabolically almost identical to procyclic forms, metacyclic forms showed characteristics of, and seemed preadapted to, differentiation into the bloodstream stage. While mitochondrial NAD + diaphorase activity and an electrochemical potential were detected in all life cycle stages, metacyclic metabolism was glucose-based and terminal oxidase activity was primarily dependent upon the trypanosome alternative oxidase with the contribution of cyanide-sensitive respiration accounting for only 20–30% of the total respiratory capacity.

Muscle mitochondrial ultrastructure in exercise-trained iron-deficient rats

To investigate effects of endurance training and iron deficiency, as well as the combination of these two conditions, on mitochondrial ultrastructure, weanling rats at 3 wk of age were assigned to iron-deficient (Fe-) and iron-sufficient (Fe+) groups. Subsequently, groups were subdivided into exercise-trained (T) and sedentary (S) groups. Electron microscopy showed subsarcolemmal and intrafibrillar mitochondria in the Fe-T animals to be enlarged with sparse cristae and vacuole-like areas compared with the other groups. An increase in the number of lipid droplets in both Fe- groups was observed. Stereological measurements revealed a 99% increase in the volume occupied by muscle mitochondria in the Fe-T animals (11.9 +/- 0.8%) over the Fe+T (5.9 +/- 0.4%) and Fe+S (6.0 +/- 0.3%) groups and a 55% increase over the Fe-S groups (7.7 +/- 0.3%). The ratio of mitochondrial surface area to tissue volume was significantly decreased only in the Fe-T group. These results indicate that the combined stresses of iron deficiency and training produce mitochondrial ultrastructural changes far greater than those of iron deficiency or training alone. Because this is also the case with the disproportion among mitochondrial enzymes, it is possible that the ultrastructural changes are indicative of morphological responses that maintain ATP turnover during exercise in iron deficiency when oxygen transport and electron transport chain activities are reduced.

Ultrastructural mechanisms of myocardial atrophy in albino rats during starvation

By means of complex morphological, morphometrical and stereological analyses the myocardium of Wistar rats was studied in full alimentary starvation during 6 days. The availability of two mechanisms of myocardial atrophy in adaptation to full starvation was revealed, these are the diminution of dimensions of parenchymatous elements and the decrease of muscle cell number. By ultrastructural investigation of cardiomyocytes the signs of structural protein synthesis decline were revealed, this is a simple myocardial atrophy. In quantitative analysis of cardiomyocytes the decrease of their number without changes in proportions of cells with different nuclei number was observed, that indicated systematic character of muscle cell elimination out of myocardium by apoptosis mechanism involvement, this is a numerical myocardial atrophy. Stereological analysis of myocardial atrophy development in conditions of full starvation determined the main regulated indices, these are the absolute total myofibril mass and mitochondrial mass and surface area.

Abnormal macrophage development in Hodgkin's disease: a morphometric study.

The development of macrophages from blood monocytes from 22 patients with untreated Hodgkin's disease and 20 normal subjects has been studied at intervals over a 6-day period of suspension culture in the presence of autologous serum and lymphocytes. Morphometric measurements were made on electron micrographs and the results subjected to multivariate and univariate analyses of variance. The cells from the Hodgkin's group showed highly significant differences from normal. The whole-cell volume, surface area, and cell membrane excess, as well as the mitochondrial volume and surface area, showed smaller increases over the period of culture, and the normal increase in mitochondrial profile numbers was not seen. It would appear that although the patients' monocytes transformed into macrophages in culture, their development was seriously deranged.

Leydig cell differentiation during maturation of the rat testis: a stereological study of cell number and ultrastructure.

In an effort to further understand the basis for the changes in steroidogenesis known to occur during sexual maturation in the rat, we examined by quantitative morphologic methods the number and ultrastructure of Leydig cells in fetal rats (days 18-20 of gestation) and in rats from days 2 to 3 of age through adult. Quantitative light microscopic analyses indicated that Leydig cell number, when expressed per unit volume of testis, was very high in fetal rat testes, fell significantly in testes of days 2 to 3 rats, and subsequently rose significantly. When Leydig cell number was expressed per testis rather than per unit volume of testis, the results indicated that testes of fetal rats and rats of days 2 to 3 contained the same number of Leydig cells; after the neonatal period, significant increases in Leydig cell number per testis occurred in concert with increases in testis weight. Quantitative electron microscopic studies revealed significant differences in the ultrastructure of fetal and adult populations of Leydig cells. For example, Leydig cells of fetal and neonatal rats contained abundant lipid, whereas Leydig cells of weeks 7 to 8 and adult rats contained little. Stereological analyses also revealed dramatic changes in smooth endoplasmic reticulum and inner mitochondrial membrane surface areas during sexual maturation, both per cell and per testis. These findings are discussed with respect to the steroidogenic capacity of the testis during sexual maturation.

Restoration effects of exogenous luteinizing hormone on the testicular steroidogenesis and Leydig cell ultrastructure.

In the present study, we explored the restoration effects of exogenous LH on Leydig cell ultrastructure and testicular steroidogenesis in rats that were deprived of endogenous LH via treatment with testosterone-17 beta-estradiol-filled Silastic implants for 10 days. Exogenous LH was supplied continuously via Alzet miniosmotic pumps at the rate of 1 microgram/h for 3, 6, 12, and 24 h or 1, 2, 4, 8, and 12 days. Testes were then perfused in vitro with medium containing 1) LH, 2) 20 alpha-hydroxycholesterol, or 3) pregnenolone substrate, which allowed us to assess LH-stimulated testosterone secretion, cholesterol side-chain cleavage activity, or the conversion of pregnenolone to testosterone, respectively. Other testes were perfusion fixed via the testicular artery for morphometric measurement of Leydig cell number and volume per testis and the surface area of Leydig cell cytoplasmic smooth endoplasmic reticulum (SER), inner mitochondrial membrane, and outer mitochondrial membrane. The results verified that Leydig cell smooth endoplasmic reticulum and inner and outer mitochondrial membrane surface areas are drastically diminished (P less than 0.05 vs. intact controls) by LH withdrawal. Also, the results verified that exogenous LH administered in situ restores Leydig cell ultrastructure and capacity to biosynthesize testosterone. However, the recovery of Leydig cell structure and steroidogenic reactions occurred at strikingly different rates upon restoration of LH after 10 days of the treatment with testosterone-17 beta-estradiol implants. For example, the restoration of testicular capacity to synthesize progesterone in response to LH stimulation or 20 alpha-hydroxycholesterol substrate was completed within 24 h. In contrast, the restoration of Leydig cell SER and testicular capacity to synthesize testosterone from pregnenolone was completed only after 8 days of continuous LH treatment (P greater than 0.05 vs. intact controls). Thus, our results show that LH rapidly restores Leydig cell post-LH receptor steroidogenic events up to and including cholesterol side-chain cleavage activity. Interestingly, there is no temporal association between the recovery of cholesterol side-chain cleavage activity and the surface area of inner mitochondrial membrane surface area. In contrast, 8 days are required to coincidentally restore SER surface area and the capacity of Leydig cells to synthesize testosterone from pregnenolone. We conclude that different cellular mechanisms are involved in the LH-dependent restoration of inner mitochondrial cholesterol side-chain cleavage activity and SER-associated conversion of pregnenolone to testosterone.

Morphometry of blood monocytes in malignant lymphoma.

Morphometric methods were used to study the ultrastructure of blood monocytes in 23 patients with Hodgkin's disease, 12 patients with non-Hodgkin's lymphoma, and 20 normal subjects. The results were analysed using both univariate and multivariate methods. Both analyses supported the interpretation that the total mitochondrial contribution to the cellular ultrastructure was less in Hodgkin's disease than in the other two groups, with smaller mitochondrial volume fractions, volumes, and surface areas being found. In the patients with non-Hodgkin's lymphoma univariate analysis suggested that monocyte nuclei were larger, with more euchromatin and a relatively smaller heterochromatin-euchromatin interface; the cell profile area was larger and the surface to volume ratio was smaller compared with normal subjects. Morphological changes have not previously been described in blood monocytes in malignant lymphoma, but they may be the counterpart of the known mononuclear phagocyte dysfunction.

Comparison of the Effectiveness of Myocardial Preservation in Right Atrium and Left Ventricle

Ten patients underwent cardiac operations during which myocardial preservation was provided by systemic hypothermia, topical cardiac cooling, and cold blood cardioplegia. The duration of ischemia ranged from 45 to 142 minutes (mean, 84.2 ± 36.2 minutes). Two serial specimens (preischemic and ischemic) were obtained from the right atrium and the left ventricle, respectively; thus, a total of 40 biopsy specimens was obtained from these 10 patients. A combination of grading of ischemic injury and stereological morphometric measurement of mitochondria was performed to assess the effectiveness of myocardial preservation. Our findings from the mitochondrial score studies (grading of ischemic injury) were as follows. In the right atrium, the average mitochondrial score rose from 0.337 ± 0.235 in the preischemic stage to 1.969 ± 0.492 in the ischemic stage. In contrast, the average mitochondrial score for the left ventricle was only elevated from 0.380 ± 0.161 to 1.353 ± 0.396. The difference between preischemia of the right atrium and left ventricle is not statistically significant, but the difference between ischemia of these chambers is significant ( p < 0.01). Our stereological morphometric studies revealed that in the left ventricle, the average mitochondrial surface area was 0.316 ± 0.046 μm 2 in the preischemic stage and 0.347 ± 0.073 μm 2 in the ischemic stage, a 9.8% increase in mitochondrial size (not significant). In contrast, the mitochondrial surface area of the right atrium showed a mean increase of 65.8%, from 0.231 ± 0.038 μm 2 in the preischemic stage to 0.383 ± 0.057 μm 2 in the ischemic stage ( p < 0.001). These data showed that with regard to ultrastructural and morphometric studies of mitochondria, there was significantly poorer preservation of the right atrium compared with the left ventricle.

Impaired adrenal reserve in the Watanabe heritable hyperlipidemic rabbit: Implications for LDL-receptor function in steroidogenesis

The cholesterol required for steroidogenesis may be provided by de novo biosynthesis or through the delivery of cholesterol by the circulating lipoproteins. By studying adrenocortical function, structure and biosynthetic capacity in an animal model devoid of the classical, high-affinity low-density lipoprotein (LDL) receptor pathway, the respective roles of de novo cholesterolgenesis and lipoprotein cholesterol delivery were investigated. The Watanabe Heritable Hyperlipemic (WHHL) rabbit lacks the LDL-receptor pathway. The activity of the rate-limiting enzyme in cholesterolgenesis, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase was 4- to 15-fold greater than normal in the WHHL adrenal gland. The basal corticosterone concentrations were normal in the WHHL rabbit; however, the corticosterone concentration increased by less than 50% of normal after an intravenous ACTH injection. Electron-microscopic evaluation of adrenocortical cells from the WHHL rabbits disclosed significantly increased mitochondrial surface area and diminished amounts of cytosolic lipid and lysosomal area. These data indicate that the mammalian adrenal gland utilizes endogenously synthesized cholesterol as well as cholesterol delivered through the LDL-receptor pathway. Moreover, in the absence of the LDL-receptor pathway, endogenously produced cholesterol is sufficient for normal basal glucocorticoid function.

Comparison of Blood Cardioplegia to Electrolyte Cardioplegia on the Effectiveness of Preservation of Right Atrial Myocardium: Mitochondrial Morphometric Study

The right atrium differs from the left ventricle in two respects during cardioplegic arrest: a higher proportion of noncoronary collateral flow is delivered to the right atrium, and the atrium is frequently excluded from topical ice cooling because of its higher position relative to the left ventricle. These factors result in early rewarming of atrial myocardium. To the best of our knowledge, the surgical literature contains no reports on whether blood cardioplegia can provide better atrial myocardial preservation than electrolyte cardioplegia. Twenty consecutive patients who underwent cardiac operations were randomly selected to receive blood cardioplegia (Group 1) or electrolyte cardioplegia (Group 2). Hypothermia was achieved by systemic cooling and continuous topical cooling with ice slush. Stereological morphometric study of mitochondria was performed on 40 biopsy specimens taken from the right atrium prior to aortic cross-clamping (preischemia) and at the end of ischemia. In Group 1, total aortic cross-clamp time was 72.8 ± 32.5 minutes. The mean mitochondrial surface area before ischemia was 0.224 ± 0.032 μ 2 and after ischemia, 0.336 ± 0.032 μ 2, a 50.0% increase in mitochondrial size. In Group 2, total aortic cross-clamp time was 69.7 ± 30.9 minutes. The mean mitochondrial surface area before ischemia was 0.205 ± 0.025 μ 2 and after ischemia, 0.439 ± 0.111 μ 2, an average increase in mitochondrial size of 114.2%. There was no significant difference between the two groups in mitochondrial size before ischemia. However, after ischemia the mean mitochondrial surface areas were significantly different ( p < 0.05). We conclude that blood cardioplegia provided better preservation of right atrial myocardium than electrolyte cardioplegia with reference to the mitochondrial morphometric study.

A comparative study of the metabolic capacity of hearts from reptiles and mammals

1.1. The metabolic capacities of reptilian and mammalian hearts have been investigated using two methods: measurement of mitochondrial enzyme activity (cytochrome oxidase) and measurement of both mitochondrial volume density and membrane surface area.2.2. The heart tissues from the reptiles and mammals showed 2-fold “weight specific” and 3-fold total organ metabolic capacity differences.3.3. Heart mitochondria from reptiles and mammals showed 2-fold differences in the activity of their enzymes per mg of mitochondrial protein yet showed very similar mitochondrial surface areas per cm3 of mitochondria.4.4. Heart mitochondria differ from liver mitochondria which have the same enzyme activities per mg of protein and the same mitochondrial surface area per cm3 of mitochondria in both the reptiles and mammals.5.5. A wide variety of reptiles and mammals both showed relationships between total heart metabolic capacity and body weight.6.6. Mammals have larger hearts than similar sized reptiles and their hearts have a greater proportion of cellular volume occupied by mitochondria.

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").