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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 14 (1990), S. 106-125 
    ISSN: 0741-0581
    Keywords: Aging ; Ultrastructure ; Liver volume ; Lysosomes ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Aging is accompanied by a myriad of changes in cell structure, function, and composition. The fact that much of the information concerning age-related alterations in cellular morphology is qualitative precludes meaningful correlations with biochemical changes in order to enhance data interpretation. The mammalian liver has been subjected to both qualitative and quantitative evaluations of hepatocyte structure as a function of aging, i.e., development, maturation, and senescence. Although these data are characterized by considerable variability and, in some instances, blatant contradictions, there exists sufficient agreement in several parameters to permit a consensus in the inbred rat model. Certainly the volume of individual hepatocytes increases with age, and many of the organelle compartments reflect this change. While old rats exhibit a high incidence of polyploidy, there is no definitive evidence to demonstrate a concomitant increase in the binuclear hepatocyte index. Several specific hepatocellular organelles undergo changes in their relative volume or surface area that appear to correlate with functional alterations. The volume density of the lysosomal compartment enlarges significantly during senescence and is accompanied by increased activities of several constituent hydrolases. The hepatic concentration of smooth-surfaced endoplasmic reticulum declines markedly with aging, as does the yield of liver microsomes and the activities of several microsomal enzymes, e.g., mono-oxygenases and glucose-6-phosphatase. However, the responses of the majority of hepatocyte organelles to aging is varied and inconsistent based on the limited data currently available.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 14 (1990), S. 179-207 
    ISSN: 0741-0581
    Keywords: Ultrastructure ; Liver disease ; Hepatitis ; Alcoholic injury ; Storage diseases ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Hepatocytes respond to injury by a few basic pathological reactions that are reflected in cell death, different types of degeneration, regeneration, or tumorous transformation. At the ultrastructural level, alterations of cell organelles can be observed in different combinations as a result of the injury, depending on the etiological agent(s) or pathological conditions developed. Nuclear bodies, dilation and fragmentation of rough endoplasmic reticulum (rer), swelling of mitochondria, and an increased number of lysosomes occur during acute viral hepatitis. The core and surface components of the hepatitis B virus can be localized in the liver cells in chronic hepatitis and in carriers. Close contact of hepatocytic and lymphocytic cell membranes were observed in chronic active hepatitis. Hepatocytes surrounded by an increased amount of collagen fibers are characteristic of cirrhosis. Loosely arranged, fine fibrils or condensed forms of Mallory bodies are pathognomic for alcoholic injury. A wide spectrum of alterations are noted after drug treatment: the proliferation of smooth endoplasmic reticulum (ser) as an adaptive phenomenon, focal or complete necrosis of the cell, inflammation, and the like. The fine structural analysis of hepatocytic inclusions in storage diseases has a differential diagnostic value. The storage of copper and other elements can be measured by x-ray microanalysis. The study of the hepatocytic differentiation in liver tumors is highly important in establishing the diagnosis and in proving the hepatocytic origin of the tumor.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 15 (1990), S. 81-96 
    ISSN: 0741-0581
    Keywords: Antibody ; GABA ; Glycine ; Acetylcholine ; Auditory system ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Over the last several years our knowledge of neurotransmitter receptors has increased dramatically as receptor types and subtypes have been identified through the development of selective antagonists, neuropharmacological studies, and radioactive ligand binding studies. At the same time major advances were made in the immunocytochemical localization of neurotransmitters and their related enzymes. However, only recently has immunocytochemistry been used to localize neurotransmitter receptors, and these studies have been limited. Four receptors have been localized in the CNS with immunocytochemistry: the nicotinic acetylcholine receptor, the beta-adrenergic receptor, the GABA/benzodiazepine receptor, and the glycine receptor. Of these the glycine receptor has been the most thoroughly characterized. Glycine receptor immunoreactivity is highly concentrated at postsynaptic sites, and the distribution of immunoreactivity appears to correlate closely with glycinergic neurons. However, immunocytochemical studies done on other receptors suggest such a distribution may not always be the case. Some receptors may not be concentrated at postsynaptic sites, and receptor distribution may not always closely fit the distribution of the respective neurotransmitter. Work is rapidly progressing on the purification of other receptors and on the production of selective antibodies which will allow immunocytochemical studies which address these and other questions.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 16 (1990), S. 93-114 
    ISSN: 0741-0581
    Keywords: Ultrastructure ; Biochemistry ; Maturation ; Comparative ; Oocytes ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This review of the anatomical, histological, biochemical, and molecular biological literature on echinoderm oogenesis includes the entire developmental history of oocytes; from their inception to the time they become ova. This is done from a comparative perspective, with reference to members of the five extant echinoderm classes; crinoids, holothurians, asteroids, ophiuroids, and echinoids. I describe the anatomy and fine structure of the echinoderm ovary, with emphasis on both the cellular relationships of the germ line cells to the somatic cells of the inner epithelium, and on the neuromuscular systems. I review the literature on the growth of oogonia into fully formed oocytes, including the process of vitellogenesis, presenting an ultrastructural analysis of the organelles and extracellular structures found in fully formed echinoderm oocytes. Echinoderm oocyte maturation is reviewed and a description of the ultrastructural, biochemical and molecular biological changes thought to occur during this process is presented. Finally, I discuss oocyte ovulation, the severing of cellular connections between the oocyte and its surrounding somatic epithelial cells.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 16 (1990), S. 202-234 
    ISSN: 0741-0581
    Keywords: Xenopus ; Meiosis ; Ultrastructure ; Intracellular signals ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Amphibian oocytes, arrested in prophase I, are stimulated to progress to metaphase II by progesterone. This process is referred to as meiotic maturation and transforms the oocyte, which cannot support the early events of embryogenesis, into the egg, which can. Meiotic maturation entails global reorganization of cell ultrastructure: In the cell cortex, the plasma membrane flattens and the cortical granules undergo redistribution. In the cell periphery, the annulate lamellae disassemble and the mitochondria become dispersed. In the cell interior, the germinal vesicle becomes disassembled and the meiotic spindles form. Marked changes in the cytoskeleton and mRNA distribution also occur throughout the cell. All of these events are temporally correlated with intracellular signalling events: Fluctuations in cAMP levels, changes in pH, phosphorylation and dephosphorylation, and ion flux changes. Evidence suggests that specific intracellular signals are responsible for specific reorganizations of ultrastructure and mRNA distribution.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 14 (1990), S. 39-45 
    ISSN: 0741-0581
    Keywords: Exocrine pancreas ; Cryofixation ; Cryomicrotomy ; Freeze-drying ; Freeze-substitution ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This paper describes and compares the morphology of a relatively complex tissue, the exocrine pancreas, prepared by state-of-the-art anhydrous cryoprocedures. Cryopreparative procedures are being used increasingly for a wide range of applications, for example, electron-probe x-ray microanalysis and immunocytochemical localization of antigenic molecules, because they preserve the composition of the specimen better than procedures involving aqueous media. Some doubts have remained concerning the morphology of cryosections and the precise identification of subcellular structures.We show that thin and sufficiently large cryosections of fresh biological tissues can be produced using commercially available hardware. The freeze-dried cryosections display high intrinsic contrast, are stable under the beam, and allow identification of intracellular fine structure.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 14 (1990), S. 348-356 
    ISSN: 0741-0581
    Keywords: Cryofixation ; Cryoprotectant ; Dimethylformamide ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Conditions for cryofixation and freeze-substitution crucial to the ultrastructural preservation of embryonic quail retina were improved. As freeze-substitution makes gentle dehydration and chemical fixation of tissue possible, the suitability of different cryoprotectants were tested in the preceding cryofixation. Additionally, different conditions for chemical prefixation were studied. In cryofixation, all of the “classic” cryoprotectants caused more or less severe tissue destruction. Only dimethylformamide (DMF) and-with certain reservations-dimethylsulfoxide (DMSO) yielded improved structure preservation. Perfusion fixation with a mixture of formaldehyde/glutaraldehyde (FA/GA) was superior to GA alone. In comparison to conventional fixation and dehydration methods, freeze-substitution yielded better ultrastructural preservation of the embryos with fewer artifacts.
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 15 (1990), S. 20-33 
    ISSN: 0741-0581
    Keywords: Catecholamines ; Ultrastructure ; Synaptic terminals ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The monoamines dopamine, noradrenaline, adrenaline, and serotonin as well as the diamine histamine have a widespread distribution in the central nervous system within synaptic terminals and nonsynaptic varicosities. In certain regions of the central nervous system the monoamines are contained in varicosities that have no synaptic specialization associated with them, suggesting a possible neuromodulatory role for some of the monoamines. The majority of monoamine labelled structures are synaptic terminals which are characterized by the presence of small, clear vesicles (40-60 nm) and large, granular vesicles (70-120 nm) within the terminal. A third population of vesicles - small, granular vesicles - which are visible only after histochemical staining, are probably the equivalent of the small, clear vesicles present after either autoradiographic or immunohistochemical labelling. Most monoamine containing terminals contact dendrites and dendritic spines and, less frequently, neuronal somata and other axons. Both asymmetrical and symmetrical membrane specializations are associated with monoaminergic terminals; however, asymmetrical contacts are the most frequent type found. These ultrastructural results indicate that monoamine containing terminals and varicosities in general share many common morphological features, but still have diverse functions.
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  • 9
    ISSN: 0741-0581
    Keywords: Lectins ; Ultrastructure ; Visual system ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Phaseolus vulgaris leucoagglutinin (PHA-L) is a plant lectin that is anterogradely transported by neurons in the central nervous system. PHA-L is selectively taken up by cells at iontophoretic injection sites and, when immunohistochemically demonstrated, labels individual neurons completely, including their dendrites, axons, and terminal boutons. PHA-L is generally not taken up by fibers passing through the injection site and, because it produces a Golgi-like staining of even very fine axons over long distances, it is sometimes possible to light microscopically reconstruct individual neurons and their entire axon terminal arbors. When prepared for electron microscopy, the PHA-L-labeled terminals are densely and completely stained, allowing their synaptic relationships to be defined. These properties make PHA-L advantageous for studying the patterns of projection and the modes of termination of select groups of neurons in their target nuclei.We used PHA-L to study the extraretinal innervation of the cat's dorsal lateral geniculate nucleus, a thalamic visual center. Although much is known about the retinal contribution to geniculate synaptic circuitry, relatively little is known about other sources of innervation, even though these provide the majority of synaptic terminals in the nucleus (Guillery: Z. Zellforsch., 96:1-38, 39-48, 1969; Wilson et al.: Proc. R. Soc. Lond. [Biol.], 221:441-436, 1984). We used both light and electron microscopy to describe synaptic circuitry from three extraretinal sources of projections to the lateral geniculate nucleus: the visual cortex, the perigeniculate nucleus, and the parabrachial region of the brainstem. Cortical terminals labeled with PHA-L were small and formed asymmetrical synaptic contacts onto small-caliber dendrites of geniculate neurons. Peri-geniculate terminals formed symmetrical synaptic contacts primarily onto small-caliber dendrites, but some synapses were also formed onto the proximal, retinorecipient portions of geniculate dendrites. Parabrachial terminals synaptically contacted the retinorecipient portions of dendritic appendages and shafts, small-caliber dendrites, and the specialized dendritic (F2) terminals of geniculate interneurons. The symmetry of the parabrachial synaptic contacts was variable and was related to the postsynaptic target. Contacts onto dendritic appendages were asymmetrical while those onto dendritic shafts and F2 terminals were symmetrical. Our data suggest that in unlabeled material these brainstem terminals would be difficult to distinguish from cortical or perigeniculate profiles. The positioning of the parabrachial input onto the retinorecipient portions of geniculate dendrites indicates that this projection is well situated to control primary retinal transmission through the nucleus, while the location of most cortical and perigeniculate innervations implicates them in secondary feedback interactions or other aspects of geniculate function.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 16 (1990), S. 2-14 
    ISSN: 0741-0581
    Keywords: Intestine ; Epithelium ; Mucous cells ; Nucleolus ; Cell kinetics ; Ultrastructure ; Protein synthesis ; RNA synthesis ; Stem cells ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This article is a summary of our work of several years on the renewal of the intestinal epithelium. A combination of ultrastructural, radioautographic, and light microscopic analyses was carried out using normal tissue and tissue affected by inhibitors of RNA and protein synthesis. Measuring protein synthesis by 3H-leucine radioautography showed that the life span of the columnar (absorptive) cells in the rat small intestine was divisible into two main phases: differentiation (from stem to functional cell) and maturation (from functional to extruding cell), each phase and its subdivisions being well defined morphologically. Differentiation involved a linear rise in the rate of protein synthesis per cell and showed at the same time heterochromatinization and silencing of RNA transcription. Data from various experiments indicated that the cells functioned from stored information (RNA), part of which came from the nucleolus, which underwent marked and characteristic ultrastructural changes. Although transcription from rDNA ceased, the nucleolus released its ribosomal material, which added to the existing protein synthesis, presumably by recruiting excess stored mRNAs. Maturation involved a nearly linear decrease of the rate of protein synthesis per cell to a characteristic low value at which extrusion took place. A gradual exhaustion of the stored RNA was indicated to be the key factor in this decrease. Ultrastructurally, maturation was associated with a gradually increasing vesiculation of rER and Golgi. The results thus imply a regulatory role of cellular protein synthesis level in renewal. This would be an epigenetic response after the genes are silenced. The nucleolus seems to play a central role in this process, and this in turn is reflected in its characteristic ultrastructural changes. The work also included new observations on the epithelium of the rat ascending colon describing a hitherto unrecognized deep crypt mucus-secretory (“DCS”) cell which is a nongoblet mature cell type apparently arising from midcrypt mitoses. In between the DCS cells, occasional slender columnar cells were seen which displayed the ultrastructural features of stem cells. These were probably reserve stem cells. We also observed nongoblet deep crypt mucous cells in the human right colon although fewer in number than in the rat. Nucleolar regulation and the presence of reserve stem cells represent new dimensions in our understanding of renewal. Electron microscopy is an essential tool in this investigation.
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  • 11
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 15 (1990), S. 2-19 
    ISSN: 0741-0581
    Keywords: Acetylcholine ; Cholinergic fibers ; Electron microscopy ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Cholinergic synapses can be identified in immunocytochemical preparations by the use of monoclonal antibodies and specific antisera to choline acetyltransferase (ChAT), the synthesizing enzyme for acetylcholine (ACh) and a specific marker for cholinergic neurons. Electron microscopic studies demonstrate that the fibers and varicosities observed in light microscopic preparations of many brain regions are small-diameter unmyelinated axons and vesicle-containing boutons. The labeled boutons generally contain clear vesicles and one or more mitochondrial profiles. Many of these boutons form synaptic contacts, and the synapses are frequently of the symmetric type, displaying thin postsynaptic densities and relatively short contact zones. However, ChAT-labeled synapses with asymmetric junctions are also observed, and their frequency varies among different brain regions. Unlabeled dendritic shafts are the most common postsynaptic elements in virtually all regions examined although other neuronal elements, including dendritic spines and neuronal somata, also receive some cholinergic innervation. ChAT-labeled boutons form synaptic contacts with several different types of unlabeled neurons within the same brain region. Such findings are consistent with a generally diffuse pattern of cholinergic innervation in many parts of the central nervous system. Despite many similarities in the characteristics of ChAT-labeled synapses, there appears to be some heterogeneity in the cholinergic innervation within as well as among brain regions. Differences are observed in the sizes of ChAT-immunoreactive boutons, the types of synaptic contacts, and the predominant postsynaptic elements. Thus, the cholinergic system presents interesting challenges for future studies of the morphological organization and related function of cholinergic synapses.
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  • 12
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 15 (1990), S. 144-154 
    ISSN: 0741-0581
    Keywords: Ultrastructure ; Spiral ganglion ; Ontogenesis ; Myelination ; TII neurons ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The development of the spiral ganglion in the cat, the rat, and the mouse was studied by electron microscopy, from fetal stages in the cat and from birth in the rodent. In the earliest stages, a single population of ganglion cells is present. Immature spiral ganglion neurons possess small perisomatic processes that seem to disappear with development, before the myelination ganglion cells are surrounded by one or two layers of Schwann cell processes. With maturation, the Schwann process increases in number around the perikaryon and its processes, which leads to the onset of myelination. The onset of myelination of the cell body processes is asynchronous. The perikaryon may be delayed in myelination by several days. Moreover, ganglion neurons from a given region of the cochlea do not myelinate simultaneously. The differentiation of two types of fibers in the intraganglionic spiral bundle and the first appearance of TII neurons occurs around birth in the cat and a few days after birth for the rat and the mouse. The distinction of TII cells is possible due to characteristic accumulation of neurofilamentous structures in the cytoplasm.
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  • 13
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 16 (1990), S. 167-173 
    ISSN: 0741-0581
    Keywords: Plunge-freezing ; Cryofixation ; Freeze-substitution ; Monolayers ; Cultured cells ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A detailed design for a simple and inexpensive variable-speed (1.0-5.8 m s-1) pneumatic plunge-freezing device is presented. Cultured cells, grown on Formvar-coated 75-mesh gold finder grids, are pneumatically driven into a stirring mixture of propane/isopentane (3:1) cooled by liquid nitrogen (LN2). Premature freezing of the sample in the cryogenic vapors above the cryogen is prevented by plunging through an entry tube into an insulating box, to which a partial vacuum is applied. The cryogenic vapors are drafted into the box at the level of the liquid cryogen by the vacuum, thereby preventing a layer of cold gas from collecting above the cryogen. To prevent the sample from thawing during transfer from the cryogen to the substitution medium, the box top is removed and compressed air is forced through a corrugated tube running the length of the box. The resulting boiling LN2 creates an atmosphere below -120°C in which the transfer can be accomplished.
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  • 14
    Electronic Resource
    Electronic Resource
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 16 (1990), S. 257-280 
    ISSN: 0741-0581
    Keywords: Oocyte ; Maturation ; Ultrastructure ; Gap junction ; Cortex ; Mammal ; Granulosa ; Actin ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Immature mammalian oocytes reside in ovarian follicles with junctionally coupled granulosa cells. When released from a currently undefined meiotic arresting influence, these oocytes resume meiosis to progress from late diplotene (germinal vesicle stage) through the first meiotic division to metaphase II. Oocytes remain at metaphase II until fertilization activates them to complete meiosis. This review summarizes ultrastructural events that occur during meiotic maturation in mammals. Developmental correlates that promise a clearer understanding of regulatory mechanisms operating to control maturation are emphasized. By use of TEM of thin sections, freeze-fracture analysis, and replicated oocyte cortical patches, we demonstrate stage-specific changes in the oocyte nucleus, reorganization of cytoplasmic organelles, correlations between oocyte maturational commitment and the junctional integrity of associated granulosa cells, and definition of the components comprising the oocyte cortical cytoplasm.
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