Quantitative Ultrastructural Investigations of Mitochondrial Development in Leishmania donovani During Transformation*

SYNOPSIS. The ultrastructure of the mitochondrion during transformation of Leishmania donovani from amastigote to promastigote stages was studied by morphometric analysis. There was a slight but significant decrease in relative mitochondrial volume (Vvli) during transformation. This decrease was linear with time for at least 27 hr. No change in relative lipid inclusion volume (Vvli) was observed during transformation. The ratio of inner to outer mitochondrial surface membrane densities (Svmii:Svmio) also remained unchanged. Although the relative number of cristae remains unchanged after transformation, there is an increase in cristae number in the portion of the kinetoplast opposite the flagellum.     

Short-term thermal acclimation induces adaptive changes in the inner mitochondrial membranes of fish skeletal muscle

In the oxidative muscles (musculi laterales superficiales) of crucian carp Carassius carassius acclimated for 6 weeks to either 5 or 25° C, the volume density and the surface density of fibres per tissue did not differ significantly between the control and experimental groups. The correlation ratio (μ²) for these values was below 50, 39·3 and 43·9 respectively. After acclimation to 5° C, the surface density of outer mitochondrial membrane per fibre increased significantly from 0·93 to 1·23m² cm⁻³ in the summer population but dropped from 0·94 to 0·67 m² cm⁻³ in the winter population. The surface density of outer mitochondrial membrane per mitochondrion increased from 3·24 to 4·52 m² cm⁻³ in summer fish. After acclimation to 25° C, the surface density of inner mitochondrial membranes per muscle fibre decreased from 4·04 to 1·79 m² cm⁻³ in summer fish and from 3·86 to 1·07 m² cm⁻³ in winter fish. The surface density of inner mitochondrial membranes per mitochondrion increased from 14·17 to 15·60 m²cm⁻³ in summer fish but dropped from 13·91 to 10·67 m² cm⁻³ in winter fish. Correlation matrices demonstrate a negative correlation of the surface density of outer mitochondrial membrane per mitochondrion with the volume density of mitochondria per fibre and temperature, suggesting cold-induced proliferation of small mitochondria. It was concluded that short-term cold acclimation increased surface area of the inner mitochondrial membranes in summer fish.     

The effect of urban ground cover on microclimate,growth and leaf gas exchange of oleander in Phoenix,Arizona

We assessed how small patches of contrasting urban ground cover [mesiscape (turf), xeriscape (gravel), concrete, and asphalt] altered the microclimate and performance of adjacent oleander (Nerium oleander L.) plants in Phoenix, Arizona during fall/winter (September–February) and spring/summer (March–September). Ground-cover and oleander canopy surface temperatures, canopy air temperatures and pot soil temperatures tended to be lowest in the mesiscape and highest in the asphalt and concrete. Canopy air vapor pressure deficits were lowest in the mesiscape and highest in the asphalt plot. Rates of net photosynthesis of all oleander plants were highest in October and May, and declined through mid-summer (June–July), when rates tended to be highest in the cooler mesiscape, particularly when water was limiting. During fall/winter, oleanders in the mesiscape produced 20% less biomass, 13% less leaf area, and had 12% lower relative growth rates (R_G) than those in the other ground covers. Lower nighttime temperatures in the mesiscape in December led to oleander frost damage. During spring/summer, oleanders in the mesiscape produced 11% more biomass, 16% more leaf area, and had 3% higher R_G than those in the other cover types. The effects of urban ground cover on oleander performance were season-specific; while oleander growth was greatest in the mesiscape during spring/summer, it was lowest during fall/winter and these plants experienced frost damage. Because all oleander plants produced >10 times as much biomass during the spring/summer, on an annual basis oleanders in the mesiscape produced 5–11% more biomass than plants in the warmer ground covers.     

Thermal compensation of peripheral oxygen transport in skeletal muscle of seasonally acclimatized trout

Seasonal changes in ultrastructure of locomotory muscle were quantified after acclimatization to natural temperature and photoperiod. Only modest changes were seen in the volume density (V(v)) of mitochondria in slow fibers ranging from 0.21 +/- 0.01 (summer) to 0.24 +/- 0.01 (winter), despite an increase in fiber size from 945 +/- 19 to 1,594 +/- 46 microm(2), respectively, resulting in a significantly greater total mitochondrial volume at low temperatures. In contrast, intracellular lipid stores showed a marked change with season, from a maximum V(v) of lipid droplets of 0.16 +/- 0.01 in winter, progressively declining through spring and summer to a minimum of 0.07 +/- 0.01 in autumn. For both organelles, the surface density reflected changes in V(v), indicating little modification of structure. Seasonal effects may dominate those of environmental temperature on mitochondrial separation, which in winter and spring fish at 4(o)C averaged 0.64 +/- 0.06 and 1.20 +/- 0.07 microm, respectively. The extracellular transport of oxygen also varies with season, the peak capillary density in autumn (2,851 +/- 88 mm(-2)) resulting in a minimum tissue supply (domain) area of 529 +/- 9 microm(2) per capillary. As a consequence, the predicted intracellular PO(2) ( approximately 2.5 kPa) is similar throughout the year.     

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.     

香樟叶肉含晶细胞季节变化研究

为探讨香樟(Cinnamomum camphora)叶肉含晶细胞超微结构的季节变化,阐明香樟叶肉中草酸钙晶体在春夏秋冬的变化规律。该研究以多年生香樟(C. camphora)叶片为材料,分别于春夏秋冬四个季节露地取样,制作超薄切片,用透射电子显微镜(TEM)观察叶肉含晶细胞超微结构的变化。结果表明:春季时香樟叶肉中只有少数细胞有草酸钙晶体,数量较少,晶体结构多为柱状晶、方晶; 夏季时香樟叶肉细胞中随机分布于液泡的草酸钙晶体明显比春季的数量多、体积大、形态丰富,晶体多为柱状晶、方晶、针晶、簇晶; 秋季时香樟叶肉细胞草酸钙晶体和夏季的类似,数量较多,形态多样,以方晶和柱状晶针晶为主,伴有晶簇; 冬季时香樟叶肉含晶细胞晶体形态为柱状晶、方晶、针晶,数量比夏季和秋季的数量略有减少。该研究结果表明在一年四季中香樟叶肉细胞液泡中均有草酸钙晶体结构存在。     

Seasonal changes in hepatocyte ultrastructure correlated with the cyclic synthesis of secretory proteins in the winter flounder (Pleuronectes americanus)

Liver tissue was sampled from flounder (Pleuronectes americanus) throughout the year with the intention of documenting changes in the ultrastructure coincident with the production and secretion of antifreeze proteins. In the winter, hepatocytes are dedicated to the production of these proteins and, in the female, also reproductive proteins. In both sexes, liver cells in the summer contain abundant lipid and glycogen stores. In the female, there is a conspicuous hepatocyte transformation from a fat-filled cell in the summer to one with well-developed rough endoplasmic reticulum in the winter. Large amounts of rough endoplasmic reticulum (11.2 mg/gm) were recovered after subcellular fractionation of female wintertime liver. The increased appearance of secretory organelles and the high number of nucleolar profiles observed in winter animals is consistent with the elevated demand for protein secretion and synthesis in both sexes. The fractional volumes occupied by lipid droplets and mitochondria were different when comparisons were made between sex and season. Females contained a greater volume of lipid than did males, and summer animals contained more lipid than those in winter.     

Improved structural preservation in freeze-substituted sporidia ofUstilago avenae—a comparison with low-temperature embedding

Summary In the present study the cellular fine structure of freeze-substituted sporidia of the phytopathogenic fungusUstilago avenae is investigated by means of thin-section electron microscopy. A conventional embedding method using Spurr's low viscosity resin is compared with the recently developed methacrylate mixtures Lowicryl^® K 4 M and HM 20 resin. Generally, freeze-substitution yields improved preservation of fine structural details of the fungus compared to previously applied conventional fixation methods. Using double fixation during freeze-substitution prior to conventional embedding the fungal membrane system (plasmalemma, endoplasmic reticulum), organelles (mitochondria, nucleus etc.) and other cytoplasmic features (ribosomes, cytoskeleton) appear well resolved and smoothly contoured. Aldehyde fixed and Lowicryl embedded sporidia ofU. avenae resemble these double fixed fungal specimens fairly closely. The complete low-temperature preparation produces visualization of distinct cellular details although contrast reversal of cellular membranes (er, mitochondria etc.) is sometimes observed. In particular, fine structure resolution is enhanced in Lowicryl HM 20 embedded fungal cells. This is due also to significant improvement in staining of the cellular membranes, cytoskeleton (microfilaments and microtubules) and Golgi apparatus-like areas, using tannic acid. In case of the fungusU. avenae, freeze-substitution in combination with mild glutaraldehyde fixation and final low-temperature embedding in HM 20 resin prove suitable for improved preservation of cellular ultrastructure.Abbreviations cw cell wall - cy cytoplasm - FS freeze-substitution - FS-A GA/OsO₄ freeze-substitution and Spurr's LV-embedding - FS-B GA freeze-substitution and Lowicryl K 4 M LT-embedding - FS-C GA freeze-substitution and Lowicryl HM 20 LT-embedding - go Golgi apparatus-like body - GA glutaraldehyde - g glycogen deposit - l lipid droplet - LT low temperature - Lowicryl LT-embedding Lowicryl low-temperature embedding - Lowicryl LT-resin Lowicryl low-temperature resin - MeOH methanol - mf microfilament - mt microtubule - m mitochondrion - mvb multivesicular body - ne nuclear envelope - np nuclear pore - npl nucleoplasm - nu nucleolus - n nucleus - OsO₄ osmium tetroxide - pl plasmalemma - pr polyribosomes - Pb-citrate Reynolds' lead citrate - r ribosome - RT room temperature - rer rough endoplasmic reticulum - Spurr's LV-embedding Spurr's low viscosity embedding - Spurr's LV-resin Spurr's low viscosity resin - t tonoplast - Uac uranyl acetate - v vacuole     

Effects of discharge and local density on the growth of juvenile Atlantic salmon Salmo salar

The study explored the combined effects of density, physical habitat and different discharge levels on the growth of juvenile Atlantic salmon Salmo salar in artificial streams, by manipulating flow during both summer and winter conditions. Growth was high during all four summer trials and increased linearly with discharge and mean velocity. Differences in fish densities (fish m^?3) due to differences in stream volume explained a similar proportion of the variation in mean growth among discharge treatments. Within streams, the fish aggregated in areas of larger sediment size, where shelters were probably abundant, while growth decreased with increasing densities. Fish appeared to favour the availability of shelter over maximization of growth. Mean growth was negative during all winter trials and did not vary among discharge treatments. These results suggest that increased fish densities are a major cause of reduced summer growth at low discharge, and that habitat‐mediated density differences explain the majority of the growth variation across habitat conditions both during summer and winter.     

Seasonal changes in zooplankton abundance in the lower Rhine during 1987–1991

Zooplankton in the River Rhine was surveyed for five years at the Dutch sampling stations, Lobith (German/Dutch border) and Maassluis (at the point of discharge of the river into the North Sea). The zooplankton abundance showed an apparent seasonal pattern at both stations, characterized by low densities during the winter period, and higher densities during the summer period, with a spring peak. Zooplankton was dominated by rotifers at both stations, although during the winter periods the contribution of copepods was considerable. The rotifers were dominated byBrachionus angularis, B. calyciflorus, Keratella cochlearis andK. quadrata; the copepods by cyclopoid nauplii; the cladocerans by small-sized species mainly belonging toBosmina. At Maassluis the relative contribution of copepods was higher than at Lobith. Furthermore, the zooplankton at Maassluis included the speciesEurytemora affinis, characteristic for estuarine conditions. In spring, the rotifer density and water temperature and rotifer density and chlorophylla concentration were positively correlated. Furthermore, both rotifer density and chlorophylla were inversely correlated with discharge. The possible role of environmental factors (water temperature, chlorophyll content, discharge and biotic factors) controlling the river zooplankton dynamics is indicated.     

Responses of ring widths and maximum densities of Larix gmelinii to climate on contrasting north- and south-facing slopes in central Siberia

An analysis was performed of the climatic responses of the radial growth of Larix gmelinii (Rupr.) Rupr. at two sites—both of which included contrasting north- and south-facing slopes—in Tura, central Siberia, with the development of ring width and maximum-density chronologies for each slope. Both residual and standard chronologies of ring widths were positively correlated with temperature from late May until mid June on all four slopes. By contrast, standard chronologies of ring widths were negatively correlated with precipitation during the winter (from October to April) and in May on the north-facing slope at site 1 and on the south-facing slope at site 2 respectively. The negative correlations with precipitation during the winter and in May on some of the slopes suggested that delayed snowmelt in early spring might inhibit the radial growth of L. gmelinii, and the effects of snow are likely to vary with topography. Both residual and standard chronologies of maximum densities were positively correlated with temperature in early July on all four slopes. Maximum densities were also positively correlated with precipitation during summer of the previous year on all the slopes. These suggest that no major differences exist in terms of responses of maximum density to climatic factors between the north- and south-facing slopes.     

A morphometric analysis of inner membranes related to biochemical characteristics of mitochondria from heart muscle and liver in mice

Mitochondria of liver and heart muscle of mice—processed in the tissue or after isolation—were analysed by quantitative electron microscopy, and their morphometric characteristics were compared with pertinent biochemical parameters. The surface density (per unit mitochondrial volume) of the inner membranes (cristae) of heart-muscle mitochondria was found to be about twice that of liver mitochondria. No significant difference was noted in the surface densities of the outer and inner boundary membranes of either heart muscle or liver mitochondria. Isolated mitochondria, metabolically transformed to the orthodox conformation, showed morphometric characteristics very similar to mitochondria from tissue samples. The surface densities of the inner membranes showed a striking correlation with the content of insoluble mitochondrial protein and the levels of succinate dehydrogenase and cytochrome aa3 reported earlier.     

Protective action of l-carnitine on cardiac mitochondrial function and structure against fatty acid stress

Cardiovascular risks are frequently accompanied by high serum fatty acid levels. Although recent studies have shown that fatty acids affect mitochondrial function and induce cell apoptosis, l-carnitine is essential for the uptake of fatty acids by mitochondria, and may attenuate the mitochondrial dysfunction and apoptosis of cardiocytes. This study aimed to elucidate the activity of l-carnitine in the prevention on fatty acid-induced mitochondrial membrane permeability transition and cytochrome c release using isolated cardiac mitochondria from rats. Palmitoyl-CoA-induced mitochondrial respiration that was observed with l-carnitine was inhibited with oligomycin. The palmitoyl-CoA-induced mitochondrial membrane depolarization and swelling were greatly inhibited by the presence of l-carnitine. In ultrastructural observations, terminally swollen and ruptured mitochondria with little or no distinguishable cristae structures were induced by treatment with palmitoyl-CoA. However, the severe morphological damage in cardiac mitochondria was dramatically inhibited by pretreatment with l-carnitine. Treatment with l-carnitine also attenuated 4-hydroxy-l-phenylglycine- and rotenone-induced mitochondrial swelling even when the l-carnitine could not protect against the decrease in oxygen consumption associated with these inhibitors. Furthermore, l-carnitine completely inhibited palmitoyl-CoA-induced cytochrome c release. We concluded that l-carnitine is essential for cardiac mitochondria to attenuate the membrane permeability transition, and to maintain the ultrastructure and membrane stabilization, in the presence of high fatty acid β-oxidation. Consequently, the cells may be protected against apoptosis by l-carnitine through inhibition of the fatty acid-induced cytochrome c release.     

Effect of different seasons on concentration of plasma luteinizing hormone and seminal quality vis-à-vis freezability of buffalo bulls (Bubalus bubalis)

Seasonal variations in semen quality, freezability and plasma luteinizing hormone (LH) levels were studied between summer and spring. Semen volume, density and initial sperm motility did not differ significantly between different seasons. Plasma LH decreased between summer and spring but the differences were, however, not significant. Pre-freezing motility did not differ significantly but post-freezing motility varied significantly (P<0.01) between seasons. Post-freezing motility was lowest during summer and highest during winter. It can be concluded that summer spermatozoa may be fragile and cannot withstand freezing stress. To increase reproductive efficiency in buffalo during summer, semen should be frozen during winter and spring and used during hot weather conditions. Seasonal variations in plasma LH levels were insignificant.     

Metazooplankton dynamics in the Salton Sea, California, 1997–1999

The dynamics of metazooplankton populations were studied over 3 years at the saline (43 g l^–1) Salton Sea, California's largest lake. Total abundance was highest in summer following late winter/early spring phytoplankton blooms. At this time, metazooplankton consisted mostly of the copepod, Apocyclops dengizicus, and the rotifer, Brachionus rotundiformis. In August or September, severe crashes in the metazooplankton populations occurred each year in mid-lake due to strong wind events which increased mixing and caused low oxygen and high sulfide concentrations throughout the water column. Larvae of the polychaete worm, Neanthes succinea and the barnacle, Balanus amphitrite were present mostly in late winter and spring. Their scarcity in summer is due in part to persistent anoxic bottom conditions that decrease adult populations and in part to predation by tilapia, an omnivorous fish that has become abundant in the lake since the 1960s. Two Synchaeta species, rotifers not previously reported from the Sea, were abundant in winter and spring and predation on these may have permitted the copepod to persist at low levels through the winter. There were two major changes in metazooplankton dynamics since 1954–1956 in addition to the appearance of the two synchaetid rotifers in the fauna. First, there are now much lower densities of barnacle and polychaete larvae in the fall, probably due to the invasion of the zooplanktivorous fish, tilapia. Second the precipitous crashes now seen in metazooplankton densities, especially the copepod, in late summer-early fall did not occur in the 1950s possibly because fall overturn events did not result in such high sulfide levels.     

Stereologic studies on mitochondrial configuration in different organs of the rat

Summary Mitochondria from different organs of the rat with configurations ultrastructurally resembling those of isolated mitochondria of known respiratory states have been subjected to Stereologic analysis. Mitochondria were examined from mossy fibers of the granular layer of the cerebellar cortex (condensed state), of the pericentral hepatocytes (orthodox state), and of heart muscle and parietal cells of the gastric fundus (transitional state). In order to study the relationship between mitochondrial compartments and the internal membrane a partition coefficient was introduced, which expresses the volume of the matrix (E_mm) and external compartment (E_ocm) respectively per unit surface area of internal mitochondrial membrane. The Stereologic parameters investigated, i.e. surface density of the mitochondrial membranes, volume density of the mitochondrial compartments and membranes, and partition coefficients generally agreed with the visual evaluation of mitochondrial ultrastructure. However, analysis of the coefficient of variation /x × 100% for E_ocm and E_mm has shown significantly greater variability in the mitochondria of the myocardium than in the gastric mitochondria, despite similar ultrastructure. It is suggested that Stereologic methods, like time-lapse cinematography, give a compound picture of configurational variation and of its plasticity.     

Frond elongation rates of shallow waterMacrocystis pyrifera (L.) Ag. In northern Baja California,Mexico

Macrocystis pyrifera (L.) Ag.frond elongation rates were measured during autumn-winter, spring and summer in a shallow water (7.5 m depth) kelp bed in Bahía Papalote, northern Baja California. Frond elongation was maximum during spring and minimum during winter. Frond growth was significantly correlated with solar radiation, and was highest in the smallest size fronds (0–2 m). Average frond elongation rate varied between 0.3–11% d^–1 during the study period. The relationship between average frond length and elongation rate followed an exponential curve with a negative slope during autumn-winter and summer, but was best described by a straight line during spring. Standard growth rates were obtained by a graphic method. Standard growth rates had intermediate values between those reported for southern California and southern Baja California.M. pyrifera growth cycle shows a different trend from what has been previously reported.     

Breeding biology of Tenagomysis tasmaniae Fenton,Anisomysis mixta australis (Zimmer) and Paramesopodopsis rufa Fenton from south-eastern Tasmania (Crustacea: Mysidacea)

The number, size and developmental stage of young in the brood pouch of female Tenagomysis tasmaniae, Anisomysis mixta australis and Paramesopodopsis rufa was recorded throughout the year. Breeding was intensive from spring till the end of autumn for the three species. Calculation of the egg ratio for each species showed that their major reproductive peaks occurred during spring and summer. A winter depression in the breeding cycle was observed for T. tasmaniae and P. rufa, but A. mixta australis ceased breeding during winter. Seasonal variation in the length of gravid females and number of young carried was evident for these three species. Females were longer in spring and summer and carried more young than in autumn and winter. A linear relationship between female length and brood size was demonstrated for each species; annual and seasonal equations were calculated for females carrying each developmental stage. The seasonal equations showed that for a female of given length fecundity was greater during spring than any other season. Natality was estimated to be highest during late spring, summer and early autumn for the three species. No seasonal variation in the size of eggs was evident for the three species. The reproduction pattern of T. tasmaniae, A. mixta australis and P. rufa appears to be very similar to that reported for the majority of iteroparous coastal temperate mysids throughout the world.     

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.     

Seasonal variation in the activity of the Leydig cells in Egyptian Nubian goat (Zaraibi) bucks

Ten Egyptian Nubian goat bucks were used to evaluate the effect of season on testicular hormonal activity and ultrastructure. Parameters were recorded for 7 consecutive weeks in the middle of the four seasons, with blood samples being collected weekly. At the end of each of these seasons, testicular biopsies were obtained surgically for histological and cytological studies. Season had a significant effect on plasma testosterone concentration, being at its lowest level (P < 0.01) during winter and spring (1.2 and 2.6 ng/ml, respectively), while at its highest during summer (10 ng/ml). The effect of season on plasma LH concentration was higher (P < 0.01) in autumn (2.9 mIU/ml) and less in spring and summer (0.4 mIU/ml). Season of the year influenced the percentage of sectional tissue area occupied by the seminiferous tubules and interstitial tissue. Seminiferous tubules occupied the majority of the testicular tissue during winter (76.6%), with the least being occupied during spring (49.8%). The thickness of the seminiferous tubules was maximal during autumn, followed by summer (53 and 36 μm, respectively). In summer the Leydig cells contained abundant smooth endoplasmic reticulum (sER), while some areas of the cytoplasm were occupied exclusively by tubular sER, arranged in parallel—indicating the highest activity of these cells. A characteristic multivesicular structure with numerous large lipid droplets and vacuoles was recorded in the Leydig cells during spring and winter, denoting low or even arrested activity of the cells. It could be concluded that season influences the activity of the Leydig cells of Egyptian Nubian bucks, and this is reflected by their ultrastructure and secretive activity.