Membrane structures ofAcholeplasma laidlawii and its virus

The main types of ultrastructures found in the freeze-fracture faces ofAcholeplasma laidlawii S 2 and its virus MV-Lg-L 172 were (1) particles 7–19 nm in diameter, mostly located in the convex cytoplasmic fracture faces. (2) small bulges or aggregates, 13–25 nm in diameter. which occupied only limited areas of both inner and outer fracture faces of some mycoplasmas, (3) numerous tiny grains and/or spikes 2–6 nm in diameter, protruding from a finely structured background, especially in the outer concave mycoplasmal fracture faces, and (4) linear structures, most probably fibrils and thicker filaments, both in the fracture faces and around mycoplasmas and viruses and connected with them. There was a high degree of structural similarity between mycoplasmal and viral membranes; no obvious significant difference was found.     

Fine structure of the chloroplast membranes ofEuglena gracilis as revealed by freeze-cleaving and deep-etching techniques

Summary The chloroplasts ofEuglena gracilis have been examined by freeze-cleaving and deep-etching techniques.The two chloroplast envelope membranes exhibit distinct fracture faces which do not resemble any of the thylakoid fracture faces.Freeze-cleaved thylakoid membranes reveal four split inner faces. Two of these faces correspond to stacked membrane regions, and two to unstacked regions. Analysis of particle sizes on the exposed faces has revealed certain differences from other chloroplast systems, which are discussed. Thylakoid membranes inEuglena are shown to reveal a constant number of particles per unit area (based on the total particle number for both complementary faces) whether they are stacked or unstacked.Deep-etchedEuglena thylakoid membranes show two additional faces, which correspond to true inner and outer thylakoid surfaces. Both of these surfaces carry very uniform populations of particles. Those on the external surface (the A surface) are round and possess a diameter of approximately 9.5 nm. Those on the inner surface (the D surface) appear rectangular (as paired subunits) and measure approximately 10 nm in width and 18 nm in length. Distribution counts of particles show that the number of particles per unit area revealed by freeze-cleaving within the thylakoid membrane approximates closely the number of particles exposed on the external thylakoid surface (the A surface) by deep-etching. The possible significance of this correlation is discussed. The distribution of rectangular particles on the inner surface of the thylakoid sac (D surface) seems to be the same in both stacked and unstacked membrane regions. We have found no correlation between the D surface particles and any clearly defined population of particles on internal, freeze-cleaved membrane faces. These and other observations suggest that stacked and unstacked membranes are similar, if not identical in internal structure.     

Effects of Photoperiod on Supramolecular Architecture of Thylakoid Membranes from a Rice Mutant (Oryza sativa Nongken 58S)

Plants of a rice mutant (Hubei photoperiod-sensitive genic male-sterile rice, Oryza sativa L. Nongken 58S) and its wild type cv. Nongken 58 were cultured in natural summer conditions in Beijing. After induction of proper photoperiods small panicle at the stem tip emerged and developed to the stage of secondary rachis-branch and spikelet primordium formation. Subsequently, part of the rice plants received long day (LD), i.e. 10 h of day-light treatment followed by 5 h of white fluorescent illumination with 1～2 Wm-2) . The others were exposed to daylight for 10 h alternating with a 14 h of dark period as short day (SD) treatment. After 10 days of the photoperiodic treatments, the chloroplast ultrastructure of the first leave below the flag leaf was examined by freeze-fracture rotary and unidirectionally shadowed electron microscopy. At anthesis stage, Nongken 58S plants with LD treatment showed complete pollen sterility, while the same plants with SD treatment exhibited normal fertility. And fertility of Nongken 58 was not affected by photoperiod treatments. The results from electron microscopic observation showed no significant effects of either SD or LD treatment on the freeze-fractured uhrastructure of thylakoid membranes in Nongken 58. No significant difference in particle density and size distribution was found on stacked and unstacked thylakoid membrane regions of the Nongken 58S-SD and those of Nongken 58 rice. However, the particle density of the endoplasmic fracture face in the staked region (EFs) and protoplasmic fracture face in the staked region (PFs) faces detected from the leaf thylakoid membranes of Nongken 58S-SD rice was significantly higher than that of the corresponding faces from Nongken 58S-LD. In some cases much more particles on EFs faces of thylakoid membranes isolated from Nongken 58S-SD rice appeared as paracrystalline particle array, indicating increases in the number of PS Ⅱ reaction centres, LHC I and Cyt b6/f per unit area of thylakoid membrane. The particle density of the endoplasmic fracture face in the unstaked region (EFu) and protoplasmic fracture face in the unstaked region (PFu) faces from unstacked thylakoid membranes of Nongken 58S-LD was less than that of the corresponding faces from Nongken 58S-SD. And the particle density of PFu faces from margin and end of the membranes of the grana thylakoids of LD-treated Nongken 58S leaves was also less than that of unstacked thylakoid membranes from SDtreated rice. In severe cases, most of the particles on endoplasmic fracture face in the unstaked region (EFu) and protoplasmic fracture face in the unstaked region (PFu) faces were even missing, indicating a decrease in the numbers of photosystem Ⅰ , LHCⅠ , Cyt b6/f and ATPase per unit area of' thylakoid membrane. The above results could further provide an augmentation for explaning the photoperiod-sensitive genic male-sterility.     

Plasmodium berghei: Architectural analysis by freeze-fracturing of the intraoocyst sporozoite's pellicular system

The technique of freeze-fracturing has been used to study the architecture of the pellicular complex of the intraoocyst sporozoite of Plasmodium berghei. The sporozoite is surrounded by three plasma membranes and a layer of subpellicular microtubules. During freeze-fracturing, each of the three membranes can split along its hydrophobic interior to yield a total of six fracture faces. The most obvious feature of each fracture face is the presence of globular intramembranous particles on the surface. The six fracture faces differ from one another in arrangement, size, and density of these intramembranous particles. Two of the fracture faces exhibit a unique arrangement of particles in well-organized parallel rows along the long axis of the sporozoite. This arrangement has not been reported in either the erythrocytic or the exoerythrocytic forms of Plasmodium spp. Another unique feature in the sporozoite revealed through freeze-fracturing is a single suture line that traverses the long axis of the inner two membranes of the parasite.     

Quantitative analysis of intramembranous particles in the membranous system of rat liver cells at different stages of development and aging

The density of intramembranous protein particles was studied by freeze-fracture. Particle density on the fracture faces of the plasmalemma and the rough endoplasmic reticulum (RER), as well as the outer and inner membranes of the nucleus and the mitochondria in rat hepatocytes were quantified. Comparison among different age groups sampled days postcoitum (dpc), days postpartum (dpp), and months postpartum (mpp) shows age-related changes in particle density in each membrane system. With the exception of the RER, particle densities increased after the 16th dpc, reached a maximum at birth, and then decreased with increasing age. Simultaneously, the number nuclear pores shows a positive correlation with the particle density of the nuclear membranes. The particle density on the membranes of the RER shows a maximum on the 16th dpc, and on the 6th dpp. Thereafter, the density of the RER decreases slightly. In all membrane systems, the density of the particles on the external fracture faces is more variable than the density of the particles on the protoplasmic fracture faces.     

Structural Changes in Membranes of Synchronized Cells demonstrated by Freeze-cleavage

FREEZE-CLEAVAGE is a new technique for studying the ultra-structure· of biological membranes, which fractures cell membranes in half, exposing two intramembranous fracture faces^1–3: the outer fracture face (OFF) and the inner fracture face (IFF). These fracture faces are partially covered with 70 Å globular particles which are thought to be unique structural components of cell membranes, formed by the association of membrane glycoproteins and lipids⁴. The 70 Å particles are dynamic structures and rapidly increase in density in the membranes of lymphocytes following exposure to mitogenic plant proteins (Scott and Marchesi, unpublished work).     

Protease digestion of membranes. Ultrastructural and biochemical effects.

(1) Nagarse, a bacterial protease, was permitted to react with sarcoplasmic reticulum, submitochondrial and plasma membranes. Gel electrophoresis indicated that all polypeptides were labile to the enzyme, and therefore must be at least partially exposed at membrane surfaces. However, hydrolysis did not proceed to completion, and in each membrane 30-50% of the original protein mass remained after extensive digestion. Gel patterns showed that remaining polypeptide fragments were in the range of 10000 molecular weight. (2) Amino acid analysis of the original protein and membrane-bound digestion product was performed. Only minor changes were observed following digestion, suggesting that the peptide fragments remaining with the membrane did not have specialized amino acid compositions. (3) freeze-fracture analysis of Nagarse-treated sarcoplasmic and plasma membranes showed that particulate structures were present, although particle density and asymmetry of fistribution between fracture faces were decreased. In submitochondrial membranes, digested membranes were indistinguishable from the original membranes in particle density and distribution. We conclude that high molecular weight polypeptides are not required for the production particulate structures in freeze-fracture images of membranes.     

AN INTERPRETATION OF LIVER CELL MEMBRANE AND JUNCTION STRUCTURE BASED ON OBSERVATION OF FREEZE-FRACTURE REPLICAS OF BOTH SIDES OF THE FRACTURE

A modification of the freeze-fracturing technique to permit observation of replicas of both sides of the fracture is described. It has been used to study mouse liver cell membrane structure. Membranes break to give two faces with three-dimensional complementarity, although there is some small-scale mismatching which is discussed. Since the two distinctive sets of membrane faces are complementary sets, they cannot be the two outside surfaces. In particular, structures (such as particles) seen on these faces are within the membrane. It is not possible from this work to say precisely where the fracture plane goes with respect to a plasma membrane, only that it must be close to the interface between membrane and cytoplasm, or at that interface. Models, consistent with the appearance of the matching replicas, are derived for three regions of the plasma membrane: (a) The nonjunctional plasma membrane, which contains many scattered particles. Except for these particles, the otherwise flat fracture face is not at variance with a bimolecular leaflet structure. (b) Gap junctions. Each of the two membranes comprising a gap junction contains a close-packed array of particles. (c) Tight junctions. Here membranes have ridges within them.     

Alteration of human erythrocyte plasma membranes by perfringolysin O as revealed by freeze-fracture electron microscopy. Studies on Clostridium perfringens exotoxins V.

When human erythrocyte membranes were treated with perfringolysin O (Clostridium perfringens theta-toxin) and examined by electron microscopy after freeze-fracture, two ultrastructural alterations were observed in fracture faces of membrane. (1) A random aggregation of intramembranous particles was seen in the fracture face of the protoplasmic half (PF face) of all membranes treated with the toxin, even if at a low concentration (40 hemolytic units/ml). On the other hand, the aggregation in the fracture face of the exoplasmic half (EF face) was observed only in membranes treated with a high concentration (3300 hemolytic units/ml) for 2 h. (2) Round protrusions and "cavities" with 30 nm in diameter were visible in EF and PF faces of membranes treated with a high concentration, respectively. These structures were always protruded toward cytoplasmic side, but did not appear to form holes through the membrane. Ring and arc shaped structures with a dark center of 26 nm and a distinct border of 5 nm in width were observed when the toxin alone was negatively stained at a very high concentration (170,000 hemolytic units/ml). These structures were also produced in the presence of cholesterol even if the toxin concentration was low.     

Asymmetric mass distribution of (Na+ + K+)-ATPase in membranes studied by freeze-fracture-etch electron microscopy

SDS-purified porcine kidney (Na+ + K+)-ATPase was studied by thin-section and freeze-etch electron microscopy. Freeze-fracturing of resealed membrane fragments shows no difference in the distribution of intramembranous particles of approx. 9.0 nm in diameter between convex and concave fracture faces. However, two types of convex face are found: FA, which shows a rather smooth background with many intramembranous particles, and FB, which shows a textured background with very few or no intramembranous particles. Etching the fractured samples further reveals that FA faces are covered with many intramembranous particles, while the etched external faces (EA) are either irregularly granulated or reveal many particles half the size of intramembranous particles. FB faces are covered with distinct pits of 9 nm or larger. The etched external surfaces (EB) are covered with many particles of intramembranous particle size. These results suggest that there are two vesicle orientations in our resealed purified membrane preparation: right-side-out, as in vivo, and inside-out. The majority of the protein mass is distributed only on one side of the membranes. Right-side-out resealed membrane vesicles after fracturing and etching show particulated FA convex fracture faces and irregularly granulated or smooth etched EA surfaces, indicating that the FA face is the protoplasmic fracture face and that the majority of the protein mass of the (Na+ + K+)-ATPase is located on the cytoplasmic half of the membrane.     

The Fine Structure of Conidia of Botryodiplodia ricinicola with Observations on Effects of Chilling

Conidia of Botryodiplodia ricinicola (Saccardo) Petrak havebeen studied, principally by freeze-etch electron microscopy.Freshly harvested conidia have a thin scaly surface layer, freeof rodlets, which covers an otherwise homogeneous-looking wallwhich is continuous with the single centrally-perforate septum.The contours of the plasmalemma are usually smooth. Nuclei andsmall vacuoles are numerous. Hydrophobic fracture faces of theplasmalemma, tonoplasts and nuclear membranes variously revealintra-membrane particles or corresponding depressions or both.Lipid inclusions are small and numerous. Compact orderly stacksof membranes are present, sometimes one in each locule of theconidium. Conidia of a strain insensitive to chilling were seento differ only in respect of the distribution of intra-membraneparticles on fracture faces of tonoplasts. Chilled and chilled-and-soakedconidia of the wild type showed fine-structural differencesfrom untreated conidia, most obviously in respect of the greatersize of some of the lipid inclusions, but also in respect offeatures of the plasmalemma which after chilling contained plasmalemmasomesand, after subsequent immersion for 15 min, showed annular depressions.Also, intra-membrane particles in some membrane systems showedaltered distribution between the two hydrophobic fracture faces.It is concluded that cell lipids and cytoplasmic membrane systemsmay be involved in the previously demonstrated chilling sensitivityof conidia of this species. Botryodiplodia ricinicola, conidia, ultrastructure, chilling effects     

Plasmodium falciparum: freeze-fracture of the gametocyte pellicular complex

Freeze-fracturing has been used to study the architecture of the pellicular complex of the gametocytes of Plasmodium falciparum. The gametocyte is surrounded by three membranes and a layer of subpellicular microtubules. During freeze-fracturing, each of the three membranes is split along its hydrophobic interior to yield a total of six fracture faces. The most obvious feature of each fracture face is the presence of globular intramembranous particles on their surfaces. The six fracture faces differ from one another in arrangement, size, and density of these intramembranous particles. In gametocytes, unlike in sporozoites, the intramembranous particles are always distributed randomly and lack any definite pattern or orientations. A unique feature of gametocytes revealed by the freeze-fracturing technique is the presence of several transverse sutures on the middle membrane that encircle the gametocyte and give it a segmented appearance.     

William Trager: An Appreciation

SYNOPSIS. Additional information on host interactions with trypanosomatid membranes was obtained from studies of a monomorphic strain of Trypanosoma brucei harvested at peak parasitemia from intact and lethally irradiated rats. Pellets of trypanosomes were fixed briefly in glutaraldehyde and processed for thin section electron microscopy or freeze-cleave replicas. Observations of sectioned material facilitated orientation and comparison of details seen in replicas. Fracture faces of cell body and flagellar membranes as well as 3-dimensional views of the nuclear membrane were studied. Cell body membranes of 80% of the organisms from intact rats contained random arrays of intramembranous particles (IMP). Aggregated clusters of particles appeared on the fracture faces of 20% of the trypanosomes. Some of these membranes had nonrandomly distributed particles aligned in distinct rows on the outer fracture face of both cell body and flagellum. Many inner face fractures of the cell body membranes had a particle arrangement similar to the longitudinal alignment of cytoskeletal microtubules. No aggregated particle distribution was seen in membranes of trypanosomes harvested from lethally irradiated rats. Replicas of trypanosome pellets also had plasmanemes as a series of attached, empty, coated membrane vesicles. These structures were found in close association with, as well as widely separated from the parasites. The shedding of these vesicles and the variation of particles in cell body membranes are discussed in light of antibody-induced architectural and antigenic changes in surface properties of trypanosomatids. The convex face of the inner membrane of the nucleus also is covered with randomly arrayed particles. More IMP were observed on the inner than on the outer nuclear membranes. Images of nuclear pores were also seen. The importance of these structures in drug and developmental studies of trypanosomes is discussed. On fracture faces of the flagellar membrane there were miniature maculae adherentes, unique to the inner fracture face and occurring only at regions of membrane apposition between cell body and flagellum. Each cluster of particles exposed by the freeze-cleave method corresponds to an electron-dense plaque seen in thin section images. However, because of a unique fracture pattern, these plaques were not revealed on the apposing body membranes, as illustrated in thin sectioned organisms.     

Alteration of human erythrocyte plasma membranes by perfringolysin O as revealed by freeze-fracture electron microscopy. Studies on Clostridium perfringens exotoxins V

When human erythrocyte membranes were treated with perfringolysin O (Clostridium perfringens θ-toxin) and examined by electron microscopy after freeze-fracture, two ultrastructural alterations were observed in fracture faces of membrane. (1) A random aggregation of intramembranous particles was seen in the fracture face of the protoplasmic half (PF face) of all membranes treated with the toxin, even if at a low concentration (40 hemolytic units/ml). On the other hand, the aggregation in the fracture face of the exoplasmic half (EF face) was observed only in membranes treated with a high concentration (3300 hemolytic units/ml) for 2 h. (2) Round protrusions and ‘cavities’ with 30 nm in diameter were visible in EF and PF faces of membranes treated with a high concentration, respectively. These structures were always protruded toward cytoplasmic side, but did not appear to form holes through the membrane.Ring and are shaped structures with a dark center of 26 nm and a distinct border of 5 nm in width were observed when the toxin alone was negatively stained at a very high concentration (170 000 hemolytic units/ml). These structures were also produced in the presence of cholesterol even if the toxin concentration was low.     

Ultrastructure of cell wall and thylakoid membranes of the thermophilic cyanobacterium Synechococcus lividus under the influence of temperature shifts

The ultrastructure of the cell wall and the thylakoid membranes of the thermophilic cyanobacterium Synechococcus lividus was studied by freezefracture electron microscopy after temperature shifts. Different fracture faces of the outer, the cytoplasmic and the thylakoid membranes were demonstrated when the preparation-temperature was in the range of the optimal growth temperature at 52°C or after fixation at 52°C. In the outer membrane of the cell wall two fracture faces with holes and 7.5 nm intramembrane particles were detected. On both the outer (EF) and inner (PF) leaflet of the cytoplasmic membrane randomly distributed particles were demonstrated. The particle density on the PF-face was approx. three times that of the EF-face. The EF-face of the thylakoid membrane exposed rows of particles with an average diameter of 10 nm. The spacing between the particle rows was 35–50 nm. This regular particle arrangement on the EF-face was demonstrated only in a few cases. Mostly the intramembrane particles were distributed randomly on the thylakoid fracture faces. The particle density of thylakoids with a random distribution was approx. in the same range both on the EF-and PF-face. The EF-particles fall into four groups of 9,10,11, and 12.5 nm. The main particle class was the 10 nm class. The PF-face exposed smaller particles with two maxima at 8.5–9 nm and 10 nm. When Synechococcus lividus OH-53s was chilled to temperatures below 30–35°C before the freeze-etch preparation a phase transition took place after the temperature shift. On the fracture faces of the thylakoid and cytoplasmic membranes particle depleted areas occurred. The size of the areas were different in both membranes and dependent on the velocity of cooling. Contrary to Synechococcus lividus OH-53s in the mesophilic Synechococcus strain 6910 the phase transition point was 15°C. The lower phase transition point may be due to a higher content of unsaturated fatty acids.Dedicated to Prof. D. Peters (Hamburg) on the occasion of the 65th anniversary of his birthday     

Ultrastructure of thylakoid membranes in C. reinhardtii: Evidence for variations in the partition coefficient of the light-harvesting complex-containing particles upon membrane fracture

The ultrastructure of the thylakoid membranes of Chlamydomonas reinhardtii was investigated using cell cultures grown under light intensities of 200 and 4000 lx, respectively. A significant difference in the size distribution of the exoplasmic fracture face (EF) particles appears upon Mg²⁺ treatment of broken cell preparations from the two light growth conditions. Particles larger than 150 Å are seen at 4000 lx only. However neither the absorption spectra of chlorophyll at 77 °K, nor the chlorophyll a/chlorophyll b ratios differ in the two cell batches. In addition, the polypeptide composition of the thylakoid membranes and the Mg²⁺ effect (spillover) on the photochemical rate of Photosystem II are the same in both conditions. We conclude that the partition coefficient between the two fracture faces of light-harvesting complex-containing particles is variable. It depends on Mg²⁺ ion concentration in the incubating medium of the membranes and on the light growth conditions of the cell cultures. Our results suggest that 60- to 80-Å protoplasmic fracture face (PF) particles containing the light-harvesting complexes can aggregate either in larger PF particles (100–120 Å) or in EF particles larger than 120 Å which also contain the Photosystem II centers. That some light-harvesting complexes are located on the PF faces is confirmed by the analysis of the BF4 mutant of C. reinhardtii lacking in chlorophyll-protein complex II. The PF faces of the BF4 thylakoids display a reduced number of particles as compared to that in the wild type.     

Fine structure of the palisade zone of the rat median eminence as revealed by freeze-fracturing

Summary Nerve terminals in the palisade zone of the rat median eminence were investigated with freeze-fracture electron microscopy. Fracture face P of the specific terminals showed two populations of intramembranous particles (IMP): a large and a small variety. The large IMP-s often formed small irregular clusters. In nerve terminals the total number of both populations of IMP-s was considerably less than that found on P membrane faces of ependymal feet. On fracture face E of the nerve terminals, the number of IMP-s was about a quarter of that seen on fracture face P.On both fracture faces of most terminals a few small round impressions (or elevations respectively) were found which may be interpreted as broken necks of either exo- or endocytotic vesicles. Neither gap nor tight junctions occurred at lateral membranes of the specific axon terminals. Similarly, no membrane specializations were observed with freeze-fracturing on membrane areas adjacent to the basement membrane. The findings are discussed in relation to a possible exocytosis mechanism of the hypothalamic releasing and inhibiting hormones.     

FREEZE-FRACTURED CHLOROPLAST MEMBRANES OF GONYAULAX POLYEDRA (PYRROPHYTA)1

The chloroplast membranes of Gonyaulax polyedra Stein were studied in replicas of rapidly frozen and fractured cells. The thylakoid EF_s face lacked the large 15–16 nm particles characteristic of plants with the light-harvesting chlorophyll a/b protein, presumably because the principal light-harvesting protein of Gonyaulax is the small water-soluble peridinin-chlorophyll-protein and the chlorophyll a/b protein is absent. As in other plants, the EF_s thylakoid fracture face carried more particles (4 ×) than EF_uface. The PF faces of the thylakoid showed twice as many particles as did the EF_s faces. No circadian differences in the number or size of thylakoid membrane particles could be detected. Three membranes comprise the chloroplast envelope in Gonyaulax. They could be clearly differentiated in freeze-fractured cells. The middle envelope membrane carried many fewer particles on both the EF and PF faces than did the other two envelope membranes. The PF faces of both the outer and inner envelope membranes showed more particles than the EF faces, as do many other membranes which have been examined.     

Phagocytosis of bacteria by polymorphonuclear leukocytes: a freeze-fracture, scanning electron microscope, and thin-section investigation of membrane structure

The changes in membrane structure of rabbit polymorphonuclear (PMN) leukocytes during bacterial phagocytosis was investigated with scanning electron microscope (SEM), thin-section, and freeze-fracture techniques. SEM observations of bacterial attachment sites showed the involvement of limited areas of PMN membrane surface (0.01-0.25μm(2)). Frequently, these areas of attachment were located on membrane extensions. The membrane extensions were present before, during, and after the engulfment of bacteria, but were diminished in size after bacterial engulfment. In general, the results obtained with SEM and thin-section techniques aided in the interpretation of the three-dimensional freeze-fracture replicas. Freeze-fracture results revealed the PMN leukocytes had two fracture faces as determined by the relative density of intramembranous particles (IMP). Membranous extensions of the plasma membrane, lysosomes, and phagocytic vacuoles contained IMP's with a distribution and density similar to those of the plasma membrane. During phagocytosis, IMPs within the plasma membrane did not undergo a massive aggregation. In fact, structural changes within the membranes were infrequent and localized to regions such as the attachment sites of bacteria, the fusion sites on the plasma membrane, and small scale changes in the phagocytic vacuole membrane during membrane fusion. During the formation of the phagocytic vacuole, the IMPs of the plasma membrane appeared to move in with the lipid bilayer while maintaining a distribution and density of IMPs similar to those of the plasma membranes. Occasionally, IMPs were aligned to linear arrays within phagocytic vacuole membranes. This alignment might be due to an interaction with linearly arranged motile structures on the side of the phagocytic vacuole membranes. IMP-free regions were observed after fusion of lysosomes with the phagocytic vacuoles or plasma membrane. These IMP-free areas probably represent sites where membrane fusion occurred between lysosomal membrane and phagocytic vacuole membrane or plasma membrane. Highly symmetrical patterns of IMPs were not observed during lysosomal membrane fusion.     

The Supramolecular Organization of Photosynthetic Membranes of the Thallus Stage of Porphyra leucosticta Thuret. (Bangiales,Rhodophyta) Visualized by Freeze-Fracture

The supramolecular organization of the thylakoid membranes of the thallus stage in the red alga Porphyra leucosticta is studied in replicas of rapidly frozen and fractured cells. Freeze-fractured thylakoid membranes exhibit only two types of fracture faces (EF and PF), because the lamellae in red algal chloroplasts are not stacked. The PF reveals numerous, tightly packed, but randomly distributed particles (density range from 2970 to 3550 particles/μm²). In contrast, the EF particles appear organized into parallel rows, the spacing of which is about 60–70 nm (about 8–9 particles occur along 100 nm of the line that is formed). Significant numbers of single EF particles are randomly distributed between the EF particle rows. The particles on both fracture faces (PF and EF) fall into two size classes: 10 to 11 nm (major size class) and 14 to 15 nm (minor size class).