Electron microscopy reviews最新文献

Tubulin, the main protein of microtubules (MTs), has the potency of forming a variety of other assembly products in vitro: rings, ring-crystals, C- and S-shaped ribbons, 10 nm fibres, hoops, sheets, heaped sheets, MT doublets, MT triplets, double-wall MTs, macrotubules, curled ribbons, and paracrystals. The supramolecular subunits of all of them are the protofilaments which might be arranged either parallel to the axis (e.g., in MTs, ribbons) or curved (e.g., in hoops, macrotubules). There is strong evidence that in the second case the protofilaments have an inside-out orientation compared to MTs. All assembly products mentioned are described structurally and their relevance to the in vivo situation is considered.

Moreover, MTs and the other assemblies undergo permanent changes. These dynamics occuring in both individual assemblies and assembly populations are discussed from the structural point of view.

Snake bite is often associated with localised soft tissue necrosis. Less frequently victims may suffer extensive muscle damage leading to rhabdomyolysis and the loss of muscle-specific protein. This review describes the organisation and structure of mammalian skeletal muscle, and its response to myotoxic venoms and to isolated pure myotoxic venom fractions. The clinical reports of muscle damage in man following snake bite are discussed, and the various classes of myotoxic toxins are introduced. Muscle damage caused by the toxins is next described, particular emphasis being placed on the correlation between muscle pathology seen at the light level and the morphological changes seen at the level of the electron microscope. Where known, those subcellular components of the muscle fibre that are especially sensitive to assault, and those components that appear to be spared, are identified. The relevance of the selective sparing of some components to the regenerative capacity of the skeletal muscle is considered.

This review discusses some of the recent developments in high resolution imaging of biological molecules. Electron micrographs of unstained biological molecules never show the resolution or contrast that would be predicted. Movements in the specimen caused by radiation damage, and possibly charging of the specimen are the most significant factors in the reduction of image contrast of these radiation-sensitive specimens. Until these limitations are overcome it is unlikely that the structures of biological molecules will be determined to the resolutions to which they are preserved. The causes of contrast loss in images are discussed in a quantitative manner and the use of crystalline paraffin as a model for radiation-sensitive specimens in general is described. Procedures for improving the contrast in images of biological molecules are described, including the new method of spot-scan imaging. Possible future developments, including high resolution imaging of single particles, are discussed.

Human enamel structural features are characterized by high resolution electron microscopy. The human enamel consists of polycrystals with a structure similar to Ca₁₀(PO₄)₆(OH)₂. This article describes the structural features of human enamel crystal at atomic and nanometer level. Besides the structural description, a great number of high resolution images are included. Research into the carious process in human enamel is very important for human beings. This article firstly describes the initiation of caries in enamel crystal at atomic and unit-cell level and secondly describes the further steps of caries with structural and chemical demineralization. The demineralization in fact, is the origin of caries in human enamel. The remineralization of carious areas in human enamel has drawn more and more attention as its potential application is realized. This process has been revealed by high resolution electron microscopy in detail in this article. On the other hand, the radiation effects on the structure of human enamel are also characterized by high resolution electron microscopy. In order to reveal this phenomenon clearly, a great number of electron micrographs have been shown, and a physical mechanism is proposed.

This review surveys the current state of knowledge relating to lipid polymorphism within both model lipid membrane and cellular membrane systems. Emphasis is placed upon the contribution of data obtained by transmission electron microscopy of freeze-fractured specimens. Some consideration is also given to the other important methods for the study of lipid polymorphism, namely X-ray diffraction and NMR spectroscopy. A detailed account of the different phases found in lipid mixtures within model membranes (bilayer, cubic or tetragonal, tubular or hexagonal) provides the background to the understanding of the factors involved in polymorphic phase transitions. The sequential steps involved in lipid polymorphism are defined from electron microscopical data and are related to the structural changes that can be detected within biological membranes. It is proposed that the fine structural changes detected at the initial stages of polymorphic transition in vivo may be highly relevant in relation to membrane fusion events, to the formation of tight junctions, and even to physiological transport processes. Since the later stages of polymorphic transition generally destroy the permeability barrier of model and cellular membranes, extensive rather than localized phase transition of the lipid bilayer is not at the moment considered to be compatible with cellular viability.

Ultrastructural observations of the cytoskeleton suggest that the connection of the intermediate filaments (IFs) to actin microfilaments (MFs) at the plasma membrane and the nuclear lamina inside the nuclear membrane link signals received at the cell periphery to the nucleus. When these connections are viewed in three dimensions using detergent extracted cytoskeletal preparations from tissue cultures or slices made from tissue, the IFs are seen to run without interruption from the cell periphery to the nucleus and back. The IFs form side to side connections with the nuclear lamina and pore complexes. The nucleus and the centrioles are supported and held suspended in these extracted cells where all organelles and cytosol have been removed. The IFs are particularly dense in the ectoplasm where they form a sheet and provide the scaffolding which maintains the shape of the extracted cells. The IFs in the ectoplasm are attached to desmoplakin at cell-cell desmosome adhesions and to MFs where the cells are attached to the fibronectin substratum possibly through integrin linkages at adhesion plaques. This was graphically shown by immunogold labelling of IF cells treated with nickel. In this way, it was possible to visualize the loss of the cell-cell connections at desmosomes and the disruption of the IF-MF connections in the ectoplasm. The MFs after losing their connections with the IFs, redistribute to cover the entire cell periphery. The nickel treatment of primary liver cell cultures lead to the loss of several functions including formation of the bile canaliculus, the ability to secrete fluorescein diacetate and the ability to take up horseradish peroxidase (HRP) by endocytosis. These observations support the conclusion that the IF-MF connections at the cell periphery provide both structural and functional polarity of the liver cells including uptake and secretion and the formation of bile canaliculi.

A battery of sophisticated techniques is now available to extract three-dimensional structural information from electron micrographs of biological macromolecules occurring in the form of single particles. One of these techniques, the random-conical reconstruction method, which allows low-dose imaging, has been recently perfected and is being used routinely for the study of ribosomal architecture. The analysis of the 40S mammalian ribosomal subunit serves as an illustration of the various steps of image processing. The use of classification combined with 3-D reconstruction provides the means to investigate variations of the macromolecular structure (deformations, conformational changes, etc.) that are caused by the specimen preparation. An example is provided by the changes in the shape of the 70S monosome of E. coli as it changes its orientation on the carbon grid. The most challenging applications of the techniques discussed are in the area of cryo-microscopy of ice-embedded specimens. First studies of single macromolecules imaged in this way have indicated that the 3-D imaging methods and, specifically, the random-conical reconstruction method, will be applicable under these conditions.

In this review emphasis is placed on the contribution of transmission electron microscopy to the analysis of spread chromosomes and nucleoids. Support is advanced for the DNA loop and rosette organization of meiotic and metaphase chromosomes and nucleoids. Extensive discussion is given to the biochemical treatments used for producing nucleoids and the effect of divalent cations and chelating agents on chromatin compactization (supercoiling). Detailed studies on nucleoids from hepatocytes are presented, with emphasis on the significance of DNA attachment to the internal nuclear matrix and to the nuclear lamina. It is firmly predicted that from the increasing knowledge of the structural organization of eukaryotic chromatin and the genome, a greater understanding of the functional roles of the various intranuclear structures will ultimately follow.