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PubMed1964

THE FATE OF MITOCHONDRIA DURING AGING IN TETRAHYMENA PYRIFORMIS.

During the growth cycle of Tetrahymena pyriformis the mitochondria undergo changes in position, number, and structure. Ciliates in the logarithmic growth phase possess elongated mitochondria which are aligned along the plasma membrane and are closely associated with the kinetosomes and kinetodesmata. Mitochondria appear to divide across the long axis at this time, resulting in two or more products. Throughout this phase of growth mitochondrial divisions keep pace with cytokinesis so that the population of mitochondria remains at essentially the minimal level. As the ciliates enter the stationary growth phase the mitochondria increase in number, become oval to spherical in shape, and some migrate into the cytoplasm. Intramitochondrial masses of various configurations appear at this time. Some of the mitochondria lying in the cytoplasm become incorporated into vacuoles. Within these vacuoles either a single mitochondrion appears or several mitochondria may be seen along with other cytoplasmic structures. Later in the stationary growth phase the contained mitochondria are dense and the tubules are more compact than normal. Various stages in disorganization of the mitochondria are observed in a single large vacuole. Cytochemical tests reveal the presence of acid phosphatase, suggesting that hydrolysis of the vacuolar contents occurs. Lipid droplets increase in number during the middle and late stationary phase of growth. These events are interpreted as being associated with the normal process of aging in T. pyriformis.

Acid Phosphatase↗
PubMed1989

Intercellular communication between rat anterior pituitary cells.

Cell-to-cell communication within the rat anterior pituitary was investigated in 60-day-old male rats with immunohistochemistry, scanning electron microscopy, freeze-fracture electron microscopy, and conventional transmission electron microscopy. A dense cytoreticular network of cytoplasmic processes from the folliculostellate cells was found to contain immunoreactive S-100 protein and was observed throughout the anterior pituitary. Nonimmunoreactive cells, which were granular, were situated in the center of each network. Almost all of the granulated cells were situated in close proximity to the folliculostellate cells. Scanning electron microscopy revealed that the gland consisted of microlobules enclosed by a basal lamina. On the surface of the microlobules were blood vessels whose branches invaded its internal structures. Cytoplasmic processes from folliculostellate cells projected outside the microlobule. Freeze-fracture electron microscopy demonstrated the presence of numerous intramembranous particles on the P-face of the plasma membrane. Scattered on the cell surface were groups of particles forming gap junctions. Meshworks of ridges which were representations of tight junctions were also observed near clusters of microvillous fragments. Clusters of particles forming small gap junctions were located between the meshworks of tight junctions. Small gap junctions were clearly observed by conventional electron microscopy between junctional complexes in a manner similar to that seen by freeze-fracture electron microscopy. Slender cytoplasmic processes of folliculostellate cells came in contact near the basal lamina and were adjoined by small gap junctions. The ratio of nongranular cells which contained gap junctions to those in which the junctions were absent was about 1:1. The size of the gap junctions ranged from 50 nm to 3 microns. No gap junctions were observed along the plasma membranes of the granular cells. The significance of an intercellular communication system within the anterior pituitary gland of the rat is to establish a mechanism for rapid transmission of information in an organ which lacks direct innervation.

Animals↗
PubMed1977

Extracellular matrix synthesis in blastula and gastrula stages of normal and hybrid frog embryos. II. Autoradiographic observations on the sites of synthesis and mode of transport of galactose- and glucosamine-labelled materials.

Pulse-chase labelling experiments and light- and electron-microscopic autoradiography were used to examine the sites of synthesis, mode of transport, and sites of deposition of galactose- and glucosamine-labelled materials in different developmental stages of normal developing Rana pipiens embryos and interspecific hybrid embryos formed by fertilizing the eggs of R. pipiens with the sperm of R. catesbeiana. In both normal and hybrid embryos, after 15-min pulse, grains are closely associated with juxtanuclear and cytoplasmic collections of membrane-bound vesicles which resemble the Golgi apparatus. In normal embryos following a 15-30 min pulse and a 60-min chase, grains are largely cleared from the cytoplasmic vesicles and deposited in the extracellular spaces or along cell surfaces where the extracellular spaces are relatively large. In contrast, arrested hybrid embryos given a 15-30-min pulse and a 60-min chase show a marked accumulation of grains over cytoplasmic structures such as the Golgi apparatus and vesicular elements in the cell cortex. Finally, early gastrula stage normal embryos are most active in the synthesis of galactose-labelled materials in cells above the dorsal lip of the blastopore, where cell migration is initiated.

Animals↗
PubMed1991

Structural and cytochemical study of the rat exocrine pancreas treated with dl-ethionine. I. Multilayered bodies and lesioned areas.

The fine structural changes and the reactivity for acid phosphatase (AcPase) and thiamine pyrophosphatase (TPPase) were studied in thin sections from rat pancreatic acinar cells exposed to dl-ethionine for 2-10 days. The cells from ad libitum and pair-fed controls exhibit occasionally 0.2-0.6 microns circular profiles showing reaction for AcPase and considered as presumptive lysosomes. At days 2 and 4 of dl-ethionine treatment the acinar cells exhibit presumptive lysosomes, autophagosomes and membrane-bounded cytoplasmic areas devoid of electron density and AcPase activity, containing scattered membranous elements. These regions were named lesioned areas. On 6th, 8th and 10th days a membrane bound anomalous cytoplasmic structure that represents a dense pile of layered membrane-like material (multilayered bodies, MB) was seen. The MBs consistently show AcPase activity and in rare instances TPPase activity. Freeze fracture studies reveal that the limiting membrane of the MBs has intramembranous particles whereas the multilayered membranous contents are devoid of such particles. The structure and disposition of the lamellae of the MBs seen in the replicas are similar to those of artificially prepared phospholipidic membranes.

Acid Phosphatase↗
PubMed1999

Morphometric studies of secretory granule formation in mouse pancreatic acinar cells. Dissecting the early structural changes following pilocarpine injection.

Secretory granule formation in pancreatic acinar cells is known to involve massive membrane flow. In previous studies we have undertaken morphometry of the regranulation mechanism in these cells and in mast cells as a model for cellular membrane movement. In our current work, electron micrographs of pancreatic acinar cells from ICR mice were taken at several time points after extensive degranulation induced by pilocarpine injection in order to investigate the volume changes of rough endoplasmic reticulum (RER), nucleus, mitochondria and autophagosomes. At 2-4 h after stimulation, when the pancreatic cells demonstrated a complete loss of granules, this was accompanied by an increased proportion of autophagosomal activity. This change primarily reflected a greatly increased proportion of profiles retaining autophagic vacuoles containing recognisable cytoplasmic structures such as mitochondria, granule profiles and fragments of RER. The mitochondrial structures reached a significant maximal size 4 h following injection (before degranulation 0.178 +/- 0.028 microm3; at 4 h peak value, 0.535 +/- 0.109 microm3). Nucleus size showed an early volume increase approaching a maximum value 2 h following degranulation. The regranulation span was thus divided into 3 stages. The first was the membrane remodelling stage (0-2 h). During this period the volume of the RER and secretory granules was greatly decreased. At the intermediate stage (2-4 h) a significant increase of the synthesis zone was observed within the nucleus. The volume of the mitochondria was increasing. At the last step, the major finding was a significant granule accumulation in parallel with an active Golgi zone.

Animals↗
PubMed1982

Epithelial cell motility: the effect of 2-deoxyglucose on cell migration, ATP production, and the structure of the cytoplasmic ground substance in lamellipodia of epithelial cells in culture.

Using a line of epithelial cells (SCCA5) derived from a spontaneous rat carcinoma, the glucose analogue 2-deoxyglucose (2DG) has been shown by time-lapse cinemicrography to produce a cessation of motility by 1 hour that can be reversed by replacement of the 2DG, and does not occur in equivalent media with or without glucose or in 2DG-containing media with added pyruvate and citrate. The effect on the cells at the edge of an epithelial island is to prevent the formation of new lamellipodia and produce a progressive retraction and condensation of lamellipodia already present. This effect of 2DG on motility corresponds with a significant reduction in the level of ATP that is partially restored after 30 minutes in the recovery incubation. Only a slight reduction in protein synthesis occurs in the presence of 2DG. The external morphology and the cytoplasmic ground substance of the cells were studied by scanning electron microscopy and high voltage electron microscopy respectively. It was found that after incubation in 2DG for 1 hour the outline of the free edges of the cells was distorted resulting in redistribution of microvilli, condensation of cytoplasm into strands, and irregular projections from the edges of residual lamellipodia. The structure of the cytoplasmic ground substance in lamellipodia from cells incubated in 2DG for 3 hours was distinctly different from that in cells incubated for 3 hours in 2DG then recovered for 25 minutes, or in cells incubated in glucose-containing medium for 3 hours. In the 2DG-treated cells the lattice-like structure evident in critical-point-dried cells was condensed into short thick strands that terminated in bulbous ends, whereas in cells recovered for 25 minutes the lattice material was elongated and tapering and the interlattice space relatively expanded. The results obtained support the concept of modulation occurring in the structure of the microtrabecular lattice component of the cytoplasmic ground substance coincident with alterations in cell function and metabolic state.

Adenosine Triphosphate↗
PubMed2002

Cytoplasmic dynein/dynactin mediates the assembly of aggresomes.

Aggresomes are pericentrosomal cytoplasmic structures into which aggregated, ubiquitinated, misfolded proteins are sequestered. Misfolded proteins accumulate in aggresomes when the capacity of the intracellular protein degradation machinery is exceeded. Previously, we demonstrated that an intact microtubule cytoskeleton is required for the aggresome formation [Johnston et al., 1998: J. Cell Biol. 143:1883-1898]. In this study, we have investigated the involvement of microtubules (MT) and MT motors in this process. Induction of aggresomes containing misfolded DeltaF508 CFTR is accompanied by a redistribution of the retrograde motor cytoplasmic dynein that colocalizes with aggresomal markers. Coexpression of the p50 (dynamitin) subunit of the dynein/dynactin complex prevents the formation of aggresomes, even in the presence of proteasome inhibitors. Using in vitro microtubule binding assays in conjunction with immunogold electron microscopy, our data demonstrate that misfolded DeltaF508 CFTR associate with microtubules. We conclude that cytoplasmic dynein/dynactin is responsible for the directed transport of misfolded protein into aggresomes. The implications of these findings with respect to the pathogenesis of neurodegenerative disease are discussed.

Antigens↗
PubMed1998

Ultrastructure of major intraglandular ducts in the parotid gland of the African mole-rat.

The parotid gland of a female African mole-rat, Tachyoryctes splendens, was examined by light and electron microscopy. A pure seromucous gland of conventional histology, its striated and excretory ducts contain prominent cytoplasmic inclusions that are lightly stained with toluidine blue. At the ultrastructural level, the inclusions are seen to consist of a light to moderately dense structureless matrix in which are suspended scattered, randomly oriented filaments that either are 4-7 nm or approx 14 nm thick. A few multivesicular bodies or putative lipid droplets are sparsely distributed in the inclusions. Even though the inclusions lack an encompassing membrane, all other formed cytoplasmic structures are excluded. These inclusions are not present in the ducts of the submandibular gland of the mole-rat. The inclusions may confer special properties on the parotid gland of this fossorial animal that permit it to cope with the exigencies of a subterranean existence.

Africa↗
PubMed1986

Some structural, biochemical and biophysical characteristics of L-929 cells growing in the presence of hyperosmotic sorbitol concentrations.

Mouse L-929 cells (a fibroblast-like line) were transferred from normal growth medium to one supplemented with 0.3 M sorbitol, doubling the normal external osmotic pressure. After a short lag phase and minimal cell death, the cells began to grow, and the growth rate reached that of controls after about one week. These chronically grown cells (S) have been compared to those of control cultures (C) with regard to general morphology, ability to reverse when returned to normal condition, water content, volume and selected metabolic parameters. S-cell cultures exhibited considerable heterogeneity but most contained vesicle-like cytoplasmic structures, sometimes in abundance. These structures do not appear to be completely bounded by membranes, but that is uncertain. S cells become larger and contain more water than C cells; however, the ratio of total water to total dry mass is indistinguishable from controls suggesting regulation at that level. S and C cells were found to be remarkably similar, on a per cell basis, with regard to their rate of respiration and the incorporation of glucose into metabolites and macromolecules. These results are interpreted in terms of current views on the composition and organization of the aqueous compartments of animal cells.

Animals↗
PubMed2000

Intracellular compartmentation of organelles and gradients of low molecular weight species.

Intracellular compartmentation of metabolites without intervening membranes is an important concept that has emerged from consideration of the metabolic inhomogeneities associated with a highly organized and structured cytoplasm within mammalian cells. This recognition is primarily due to the development of experimental approaches to measure metabolite or ion concentrations at specific subcellular sites, thereby providing a means to study concentration gradients within the aqueous cytoplasm in intact cells. The presence of mitochondrial clusters has been shown to create gradients of low molecular weight species, such as O2, ATP, and pH, with important implications for substrate supply for function and regulation of cellular processes. Moreover, the existence of kinetically distinct precursor pools has been shown to result in functional compartmentation of biochemical pathways, such as DNA replication and carbohydrate metabolism. The creation of these specialized microzones of metabolism in accordance with their association with cellular organelles or membranal structures may be integral to normal function and regulation of adult mammalian cells.

Adenosine Triphosphate↗
PubMed1990

Intracellular trafficking of two major Epstein-Barr virus glycoproteins, gp350/220 and gp110.

The processing and intracellular localization of the two predominant Epstein-Barr virus glycoproteins expressed in late lytic infection were investigated. Immune light or electron microscopy of frozen fixed sections revealed that gp110 colocalized to the endoplasmic reticulum and to the nuclear membrane with the endoplasmic reticulum-resident protein, heavy-chain-binding protein (BiP), while gp350/220 accumulated in low abundance in the endoplasmic reticulum and was present in higher abundance in cytoplasmic structures presumed to be Golgi and in plasma membranes. Consistent with endoplasmic reticulum and nuclear membrane localization, the bulk of gp110 was sensitive to endoglycosidase H, indicating high-mannose, pre-Golgi, N-linked glycosylation; while consistent with Golgi and plasma membrane localization, gp350/220 was mostly resistant to endoglycosidase H because of complex N- and O-linked glycosylation. gp350/220 was as abundant in extracellular enveloped virus as in the plasma membrane but was much less abundant or undetected in internal cytoplasmic or nuclear membranes. In contrast, gp110-specific antibodies did not label extracellular or intracellular virus. These data indicate that the major antigenic components of gp110 are not incorporated into or are occluded in virions and that gp350/220 is added to virus in cytoplasmic transit through a process of de-envelopment and re-envelopment at the plasma membrane or at post-Golgi vesicles. Consistent with cytoplasmic de-envelopment and re-envelopment at the plasma membrane was the finding of some free nucleocapsids in the cytoplasm of cells with intact nuclear membranes and nucleocapsids which appeared to bud through the plasma membrane.

Animals↗
PubMed2003

Evolution without speciation but with selection: LUCA, the Last Universal Common Ancestor in Gilbert's RNA world.

This is not an attempt to analyze the Last Universal Common Ancestor (LUCA) to understand the origin of living systems. We do not know what came before Gilberts' RNA world. Our analysis starts with the RNA world and with genes (biological replicators alla Dawkings) made up of RNA proteins with enzymatic catalytic functions within units that are not yet modern cells. We offer a scenario where cellular entities are very simple and without individuality; they are only simple primary units of selection (the first level of selection) in which replicators compete in the most Darwinian manner, totally deprived of cooperation and interactions among genes. The information processing system of this RNA world is inaccurate and inefficient when compared to that found in organisms that came later. Among the "genes" and the entities that harbor them, high mutation rate was the most prevalent source of variability and the only inheritance was through lateral gene transfer of mobile elements. There were no chromosomes or any other genomic organization. As millions of years accumulated, complex and organized biological structures and processes evolved thanks to the variability mustered up mostly by lateral gene transfers and mutations. With micro- and mini-satellites, lateral gene transfers became indispensable devices of selection to mold variability. Competition and Darwinian selection gave way to a new transition in evolution, one I consider ineluctable, in which cooperation among interactive genes prevailed for the sake of higher fitness. Compartmentalization constituted a major transition in evolution that spurted new types of genome organization. Minichromosomes is one of these; cellular membranes and cytoplasmic structures completed the picture of the primitive cell. However, the much talked about phylogenetic tree does not exit in that ancient LUCA. The tree has no organism at its base; only clusters of genes evoke a fragile beginning for the increasingly complex cell types that were to emerge later.

Animals↗
PubMed1957

A study of hemosiderosis with the aid of electron microscopy; with observations on the relationship between hemosiderin and ferritin.

Hemosiderin deposits in rats and in man were studied and compared by means of electron and light microscopy. Typical, isotropic, iron-positive hemosiderin granules were found to contain innumerable, closely packed, electron-dense particles, embedded in matter that was much less dense to electrons. Similar dense particles were often scattered diffusely through the cytoplasmic matrix of cells containing hemosiderin granules. In cells of proximal convoluted tubules of rats given repeated intraperitoneal injections of hemoglobin the hemosiderin granules contained dense particles with a mean diameter of 55 A, and with a size-frequency distribution that indicated uniformity. These particles corresponded in size to the iron micelles of ferritin molecules. There was less uniformity of particles in hemosiderin granules situated in liver and reticulo-endothelial cells of rats that had been given a diet containing ethionine. The dense aggregates representing hemosiderin granules were often situated inside discrete cytoplasmic organelles that were bordered by membranes, and sometimes contained "cristae"; and often the membranous borders were markedly disrupted. The term "sidersomes" is proposed for these specialized cytoplasmic structures which may be derivatives of mitochondria, and apparently play a part in the formation of hemosiderin. Ferritin was crystallized from the livers and kidneys of the hemosiderotic rats with ease, but could not be crystallized from comparable quantities of liver and kidney tissue of untreated control rats. Specimens from the liver and spleen of a patient with advanced hemosiderosis, obtained at an operation, were also studied. In liver and reticulo-endothelial cells many particles with diameters of about 60 A were scattered through the cytoplasmic matrix. By contrast, hemosiderin granules in the same cells contained particles that varied considerably in size. In representative granules, examined at high resolution, the size-frequency distribution of particle diameters displayed a periodicity consistent with the presence of small, uniform subunits. Electron micrographs of ferritin, isolated from the spleen of the same patient, provided confirmation for the inferences that the dense particles observed inside cells are iron micelles, and that ferritin is probably a component of hemosiderin.

Animals↗
PubMed1991

Chromatin motion in neuronal interphase nuclei: changes induced by disruption of intermediate filaments.

Motion of nucleoli within interphase nuclei, known as nuclear rotation, may be used as a measure of motion of chromatin domains within the global confines of the nucleus. Mechanisms by which chromatin domains are transposed remain enigmatic. It has been established that nuclei are anchored by a network of intermediate filaments, structural proteins which share epitopes with nuclear lamins and possibly representing a constraint on nuclear rotation. It is postulated that selective removal of this constraint, by acrylamide, would result in increased chromatin motion. Mean rates of nucleolar displacement were quantified in neurons, in vitro. Nuclear rotation increased from a mean control rate of 0.102 +/- 0.002 micron/min (n = 52) to a maximum mean rate of 0.207 +/- 0.026 micron/min (n = 11), after 23 hr of exposure to 4 mM acrylamide. Despite this significant increase in motion of intranuclear domains, cytoplasmic structures in the immediate juxtanuclear area did not exhibit increases in rates of motion. Immunocytochemistry was used to visualize cytoskeletal structures and to assay selective disruption of neurofilaments by acrylamide. Increased rates of chromatin motion coincided with breakdown of the intermediate filament network. Ultrastructural analyses showed that the increase in chromatin motion induced by acrylamide was also associated with a significant (P less than 0.005) change in the thickness of the nuclear lamina, decreasing from 20.9 +/- 5.10 nm (n = 159) in controls to 18.9 +/- 3.1 nm (n = 148), to 19.5 +/- 3.6 nm (n = 240) and to 16.1 +/- 4.4 nm (n = 103) at 4, 8 and 22 hr exposure, respectively. Moreover, the number of mitochondria per unit area changed significantly (P less than 0.0001) with exposure to acrylamide, increasing from 9.1 +/- 2.2 mitochondrial profiles in controls to 16.5 +/- 5.3 profiles after 22 hr exposure to acrylamide. Distribution of other cytoskeletal components, actin and microtubules, was not altered and does not appear to play a significant role in the observed increase in rates of nuclear rotation. We conclude that the removal of the damping effects on chromatin motion normally imposed by the nuclear lamina and by intermediate filaments results in increased chromatin motion.

Acrylamide↗
PubMed1992

A morphological study of the rabbit corneal assay.

We present a morphological study of the rabbit corneal assay. The effects of corneal pockets filled with methylcellulosis carriers with and without basic fibroblast growth factor (bFGF) were studied 4, 7, 11, 14 and 21 days following surgery. In the controls, strongly vacuolated keratocytes are present at the borders and especially at the bottom of the pocket. A large number of such cells can also be found in the region between the pocket and the epithelium. These cells obviously undergo necrosis. A few vacuolated keratocytes with poorly structured cytoplasm can even be found in the stroma between the pocket and the limbus. There are also changes in the epithelium. It is thinner than normal and the cells are longitudinally oriented. Construction of the pocket sometimes results in a fissure extending a variable distance from the bottom of the pocket to the limbus. Basic FGF induced blood-vessel growth from the limbus into the cornea. At least until day 7 this is accompanied by high vulnerability of the vessels, and changes in the pattern and structure of the extracellular matrix. Neutrophilic granulocytes, monocytes and a few mast cells migrate in front of the vessels. As in the controls, vacuolated and necrotic keratocytes are present within the stroma. From day 11 onward, many more vessels can be found in front of than behind the pocket, often located adjacent to vacuolated keratocytes. We suggest that an angiogenic substance placed in the corneal pocket is not the only angiogenic stimulus in the test, but acts together with many stimuli caused by the surgical manipulations.

Animals↗
PubMed2000

Transient structure of the amyloid precursor protein cytoplasmic tail indicates preordering of structure for binding to cytosolic factors.

The cytoplasmic tail of the amyloid precursor protein (APP) appears to play two important roles in the cell through participation in intracellular signaling and proteolytic processing of APP. Hence, knowledge of the structure of the 47 residue cytoplasmic tail of APP is important for understanding the molecular interactions involved in normal cell function as well as in the pathogenesis of Alzheimer's disease. Multidimensional solution NMR spectroscopy has been applied to examine the structural features of a 49-residue peptide (APP-C) containing two N-terminal residues (GS) and the APP cytoplasmic tail, over the pH range of 4.2-7.1. Although the peptide does not adopt a stable folded structure, regions of unstable structure exist over the pH range examined and have been characterized by a combination of H(alpha) chemical shifts, NOE analysis, and (3)J(HNH)(alpha) coupling constants and by identification of transient hydrogen bonds between amide protons and titrating carboxylate groups. These studies extend the work of others [Kroenke et al. (1997) Biochemistry 36, 8145-8152] by identifying an additional nascent helix and a hydrophobic cluster within the N-terminal 20 amino acid residues and by further characterizing the TPEE turn as a helix capping box. The transient structure of APP-C provides insight into the importance of preordering of this cytoplasmic tail in governing specificity and affinity for cytosolic binding partners.

Adaptor Proteins, Signal Transducing↗
PubMed1998

In situ localization of paclitaxel binding structures: Labeling with a paclitaxel fluorescent analogue.

Microtubules are a major component of cell cytoskeleton. Microtubules constitute the cellular target of paclitaxel. The interaction of paclitaxel with microtubules causes an increase in tubulin polymerization and microtubules stabilization, leading to a G2/M phase cell cycle arrest and cell death by apoptosis. Three paclitaxel fluorescent analogues were prepared by introducing fluorescein or BODIPY moiety onto the 2' of the 7-carbon. These were then used to study the interaction of paclitaxel with its cellular binding site and the microtubule network was visualized directly by fluorescence microscopy. Free paclitaxel was able to inhibit 7-FITC paclitaxel binding, demonstrating that both products bind to the same site and possess similar biological properties. Using the carbon 7 derivatives, microtubules were labeled as cytoplasmic fibers extending centripedly. A few other cellular components such as nuclear membrane, nucleoli and some other cytoplasmic structures were also labeled. The labeling intensity was reduced by preincubation with free paclitaxel. The interaction of paclitaxel with microtubules was also investigated using flow cytometry. No binding of the 2'C derivative was detected, confirming that a free 2'C is a pre-requisite for paclitaxel microtubules interaction.

Journal Article↗
PubMed1987

Effects of iron-, manganese-, or magnesium-deficiency on the growth and morphology of Euglena gracilis.

Iron-, manganese-, or magnesium-deficiency has been induced in Euglena gracilis. Each arrests cell proliferation, decreases the intracellular content of the deficient metal, and increases that of several other metals. Light and electron microscopy of stationary phase cells reveal that Fe-deficient (-Fe) cells are similar in size and shape to control organisms. Magnesium-deficient (-Mg) cells, however, are larger, and approximately 14% are multilobed, containing 2 to 12 lobes of equal size emanating from a central region. Individual (-Mg) cells and each lobe of multilobed cells contain a single nucleus. Manganese-deficient (-Mn) organisms are morphologically more heterogeneous than (-Fe) or (-Mg) cells. Most are spherical and larger than controls. Approximately 15% are multilobed but, unlike (-Mg) cells, contain lobes of unequal size with either zero, one, or several nuclei present in each. Nuclei of (-Mn) cells differ in size and shape from those of control, (-Fe), or (-Mg) cells. All three deficient cell types accumulate large quantities of paramylon. Other cytoplasmic structures, however, appear normal. Addition of Fe, Mn, or Mg to the respective deficient stationary phase cultures reverses growth arrest and restores normal morphology. The results suggest that Fe-, Mn-, and Mg-deficiencies affect different stages of the E. gracilis cell cycle.

Cell Nucleus↗