Kaarina Pihakaski

Wintertime changes in the ultrastructure and metabolism of the microsporangiate strobili of the Scotch pine

Terminal buds of Pinus silvestris L. containing microsporangiate strobilus primordia were collected once a month throughout the winter. The electron microscopic studies indicated that in October and December, the cells of the strobili contained a large number of vacuoles, a portion of which was supposedly autophagic, and stacked rough endoplasmic reticulum. By February, the amount of these had decreased, and instead, a large population of dense bodies was visible. Additional phenomena, characteristic at this state, were the occurrences of highly uneven contours of the plasmalemma and of inclusions of various kinds between the plasmalemma and the cell wall. In March, autolysis was visible in a portion of cells outside the sporangia. In the sporangia the ground cytoplasm was thin but the number of organelles was increasing. In the April collections, cell divisions were visible. The amount of protein per dry weight increased during the winter reaching a peak in February. The activity of RNases, having optima of pH 5.0 and pH 7.5, was measured in two successive years. Both series showed a period of high activity during the middle of the winter. The exact timing of this period depended on the year in question. On the basis of these observations, the dormant period of the microsporangiate strobili of the Seotch pine is divided into three sub-periods. It is also suggested that the definition of dormancy of these structures should include a mentioning of alterations in the metabolical machinery of the cells.

Intramembrane particles and filipin labelling on the membranes of autophagic vacuoles and lysosomes in mouse liver

SummaryMorphologically detectable protein (intramembrane particles) and cholesterol (filipin labelling) in the membranes of autophagic vacuoles and lysosomes were studied in mouse hepatocytes using thin-section and freeze-fracture electron microscopy. Both isolated autophagic vacuoles and lysosomes, and intact tissue blocks were used due to the facts (i) that lysosomes are difficult to recognize in freeze-fracture replicas of intact hepatocytes, and (i) that filipin penetration into the tissue blocks is unsatisfactory. Intramembrane particle density was low in the membranes of early autophagic vacuoles (defined as round-shaped vacuoles in which an inner membrane parallel with the outer limiting membrane was clearly visible). The lysosomal membranes contained considerably more intramembrane particles. Particle-rich lysosomes or other vesicles were observed to fuse with the early autophagic vacuoles. The membranes of nascent autophagic vacuoles with morphologically intact contents were usually not labelled by filipin, whereas the membranes of all other autophagic vacuoles and lysosomes were heavily labelled. The increased cholesterol in the membranes of slightly older autophagic vacuoles is presumably derived from cholesterol-rich lysosomes or other vesicles fusing with the vacuoles and from the degrading organelles inside the autophagic vacuoles.

A study of the ultrastructure of the shoot apex and leaf cells in two liverworts, with special reference to the oil bodies

SummaryIn this investigation attention has been paid to the general ultrastructure of the shoot apical and leaf cells in the liverwortsBazzania trilobata andLophozia ventricosa but especially to the different developmental stages of their oil bodies. These species have been chosen because their oil bodies differ from each other in size and shape.The appearance of the different organelles, nucleus, chloroplasts, mitochondria, ER, and Golgi bodies, are in their main features the same as those of higher plants described in the literature. The dark cytoplasm seen in the leaf cells ofLophozia in the vicinity of the oil bodies but without any surrounding membrane when fixed in double fixative 2, seems to be specific to this species. On the other hand, granular dense bodies were visible in the cells of the shoot apex ofBazzania, which shrank in size as the development of the oil bodies proceeded and were lacking in the mature leaf cells.In both species investigated, the oil bodies have the same component parts: (1) an outer membrane enveloping the whole body, (2) inside this, a granular stroma layer of varying thickness enveloping (3) specific globules of varying size and number, each of which is surrounded by (4) a thin inner membrane (Fig. 28).The oil bodies develop in at least two ways and usually in one way for each species. InBazzania they seem to develop from vacuole-like formations in the shoot apex or in the leaf primordia into which substances have segregated. InLophozia they seem to originate by aggregation and fusion of lipid bodies.

Ultrastructural changes arising from freezing of leaf blade cells of cold acclimated rye (Secale cereale)

Summary The survival at sub-zero temperatures of leaf blade cells of rye (Secale cereale cv. Voima) that had been cold-acclimated at 5 °C for 4 weeks was determined by measuring the efflux of ninhydrin-positive substances: 50 % of the cells were dead at - 8 °C (LT50) and none survived at -16 °C or below. Specimens for transmission electron microscopy were prepared from leaf blade segments by fast freezing followed by freeze-substitution in acetone with OSO4 fixation. In specimens from plants maintained at 5 °C, chloroplasts and mitochondria were well-structured and the endoplasmic reticulum and dictyosomes easily discernible. Many membrane bound vesicles, ranging in cross sectional diameter from ~ 60 nm upwards were seen in the cytoplasm. Sometimes stacks of membrane strands running parallel to the plasmalemma for up to 2 µm were also observed. Upon freezing to -16 °C, the cells were severely dehydrated and distorted, the vacuoles severely shrunken, and the cytoplasm and mitochondria disorganized; cytoplasmic vesicles and stacks of membrane strands seen in the controls were absent. Upon freezing to - 8 °C, some cells were as disorganized as those at -16 °C, although the cell wall did not always adhere to the collapsed protoplast; others were relatively intact and some showed evidence of intracellular ice crystal formation.

Quantitative seasonal variation in mitochondrial ultrastructure of mesophyll cells ofDiapensia lapponica L. with reference to some effects of fixative osmolality

SummarySeasonal changes in the mitochondrial ultrastructure were examined in palisade parenchyma cells of a tuft-formingDiapensia lapponica L. collected at monthly intervals in Northern Finland. Quantitative analyses to measure volume and surface densities were conducted during different periods of growth (stages of growth, acclimation, winter period and deacclimation) in the annual cycle.The volume density was highest in the summer and lowest in the spring; the difference was significant with both fixatives used GA and GA/FA. The largest membrane area (the mitochondrial outer membrane and the cristal membranes together) was observed in the summer and autumn, and was significantly less in the winter and spring. This correlated with fewer mitochondria in the spring and a smaller number of cristae in the winter and spring. In the material fixed in GA/FA the distribution of length/width ratios of mitochondria was relatively uniform in all seasons. However, the mitochondrial ultrastructure had the most varied appearance during the winter. Hypertonie GA/FA solution did not cause significant differences either in the ultrastructure or the volume and surface densities of the mitochondria.

Effect of denucleation and UV-irradiation on the subcellular morphology inMicrasterias

SummaryThe effects of the elimination of the nuclear control on the ultrastructure of the green algaMicrasterias torreyi. Bail, have been studied by using centrifugation for denucleation and lethal dose of UV-light. Centrifugated anucleate cells were fixed 7 and 26 hours after the treatment and the UV-treated cells 4 and 8 hours after the irradiation. Although both treatments eliminate the nuclear control and the treated cells resemble morphologically each other, yet there are differences in ultrastructure suggesting that they are also brought about by other factors than the presence of nucleus. Both the treatments cause accumulation of cell wall material in the tips of lobes. The cell wall shows unusual secondary thickening with electron dense spots embedded in the matrix. The denucleation retards the functional cycle of Golgi apparatus and the production of vesicles has stopped in the 26-hour-denucleated cells. It is possible that flat vesicle production is totally absent in denucleated cells.First the UV-treatment seems to activate the function of Golgi apparatus but later on the vesicle production almost stops. It seems to eliminate the production of large vesicles but not that of dark vesicles.Both the treatments cause deterioration of ER membranes and polysomes, and in consequence, probably inhibit protein synthesis.Unlike UV-irradiation, denucleation appears to destroy the microtubule system. Mitochondrial cristae have almost entirely vanished within 26 hours after denucleation. Effect of denucleation and UV-irradiation on the subcellular morphology inMicrasterias.