Electron Microscope Study of the Lathyritic Rat Aorta

Six weanling rats were fed a diet containing 0.4 per cent BAPN fumarate and sacrificed after 5 to 33 days on the diet. The ascending aortae were fixed with OsO₄, embedded in methacrylate and araldite, sectioned, stained with lead hydroxide, and examined with the electron microscope. The descending thoracic aortae were examined by light microscopy. Compared with pair-fed controls, the experimentals showed definite changes which became more marked as the disease progressed. The wall became thicker with wider interlaminar spaces, radial orientation of the smooth muscle cells, progressive loss of desmosomes, and progressive increase in a dense, finely stippled material that coated the edges of the elastic laminae and extended outwards between the muscle cells and separated the ends of these cells from the laminae. This stippled material occurred at the same sites as the increase in PAS-positive and azan-positive material seen with the light microscope. There was an increase in subendothelial and interlaminar collagen, and electron microscopy clearly showed that the cells were smooth muscle and not fibroblasts. The possible bearing of the morphological changes on the formation of aortic aneurysms is discussed.     

An Electron Microscope Study of the Rat Ovum

This paper reports on the fine structure of rat oocytes at stages before ovulation, during maturation, fertilization, and early cleavage. The study includes parallel observations on light and electron microscope preparations with attempted correlations. The follicular cells of the ovarian egg are described as sending long processes through the zona pellucida to the egg surface where they mingle with thin projections from the egg itself. No open communication between follicle cell cytoplasm and egg cytoplasm was observed. During maturation and fertilization both types of processes are withdrawn from the zona. The germinal vesicle and later the pronuclei of the fertilized egg are characterized by numerous large nucleoli. These have the form of thick walled vesicles with diameters as great as 8 to 10 µ. The wall is dense in the EM image and appears to consist in part of small granules. The cytoplasm shows several inclusions including mitochondria of usual form and a Golgi component which has the typical fine structure and the distribution described by earlier light studies. Small dense particles, presumably RNP particles, are distributed throughout the cytoplasmic matrix and show no preference for membranes. The endoplasmic reticulum of the oocyte is represented by a scattering only of vesicles, but begins a more extensive and elaborate development with the onset of segmentation. One inclusion of the ooplasm, similar in size to mitochondria, receives special attention. It is a vesicular structure, containing a large number of small vesicles (10 to 50 mµ in diameter) and frequently a central density or nucleoid. They are referred to as multivesicular bodies. Such bodies are found in small number in the ovarian egg, but increase greatly in number during maturation and fertilization. It appears from the micrographs of eggs in these latter stages that these vesicular bodies break down and liberate their content of small vesicles to the surrounding ooplasm. Comments are provided on the apparent significance of the various observations.     

An Electron Microscope Study of Basophile Substances of Frozen-Dried Rat Liver

Small pieces of liver from rats subjected to different dietary regimes were fixed by freeze-drying, and postfixed by in vacuo heating and denaturation with alcohol. Specimens were digested with ribo- or deoxyribonuclease, and stained with gallocyanin-chromalum, azure II, the Feulgen procedure or alcoholic platinic tetrabromide. Some specimens were reserved as controls of the effects of enzyme treatment. Stained and unstained specimens were embedded in methacrylate and examined by light and electron microscopy. Basophilic and Feulgen-positive substances, after contact with watery reagents, were found by electron microscopy to exist as small dense granules embedded in a less dense homogeneous matrix, forming the walls of submicroscopic vacuoles. These granules were absent after digestion with nucleodepolymerases. In specimens (unstained, or stained with platinic tetrabromide) which had not passed through water, the dense (basophile) substances in nuclei and cytoplasm were found to exist, not as granules, but as ill defined submicroscopic concentrates which blended imperceptibly into the homogeneous matrix of the vacuolar walls. Objections to the use of stains for improving contrast conditions in electron microscopy of tissues are discussed, and it is concluded that the reagents do not necessarily produce the observed increases in contrast by selectively stabilizing certain structures. The concept of microsomes as pre-existing distinct morphological entities in intact (unhomogenized) cells is thought to be inconsistent with the distribution of basophile substances in frozen-dried liver.     

Electron Microscope Study of Lens Fibers

Comparative electron microscope investigations on sections of the lens cortex of the normal, mature rat, rabbit, monkey, and the normal calf reveal similar patterns of intracellular organization. The superficial lens fiber contains a nucleus, mitochondria, endoplasmic reticulum, dense granules, Golgi complex, and a quantity of small structures of low opacity which appear as filamentous and spherical configurations. Variations in number, distribution, and spatial arrangement of cytoplasmic elements in lens fibers are described. These changes in the pattern of cytoplasmic organization are concomitant with development of fibers and their displacement towards the center of the lens. Structural details of the various zones of the lens epithelium and the lens fibers are compared.     

Electron Microscope Study of the Human Neuromuscular Junction

A preliminary electron microscope study of human neuromuscular junction is presented. The biopsy material was taken from the palmarus longus, and fixed routinely in osmium tetroxide and embedded in methacrylate. The structure of the motor endings and the relationship of the synaptic vesicles to the axolemmal membrane are described. The synaptic clefts are filled with an homogeneous material in continuity with the basement membrane covering the muscle fiber. The subneural apparatus is described, and special attention is paid to a vesicular component present in the sarcoplasm of the junctional area, which differs from synaptic vesicles and is presumed to be a derivate of the sarcoplasmic reticulum.     

Electron Microscope Studies of Nuclear Extrusions in Pancreatic Acinar Cells of the Rat

This paper describes "blebs" protruding from the surface of the nucleus into the cytoplasm. The "blebs" are separated from the cytoplasm by 2 membranes which are continuous with the outer and inner nuclear membranes. The "blebs" contain 3 structurally distinct substances. Two of these substances (β and γ substances) are similar to extranucleolar karyoplasm and nucleolar material. The other substance (α substance) is present in every "bleb," but it cannot be readily compared to a recognizable nuclear structure. Cytoplasmic vesicles are described that are apparently different from the Golgi vesicles or the vesicular component of the ergastoplasm. It is suggested that these vesicles may be of nuclear "bleb" origin. A dark karyoplasmic zone extending from the region of the nucleolus into the nuclear "bleb" is shown. This zone may be similar in some respects to the preformed pathway ("Leitbahn") described by Altmann (3) and Hertl (28) and could reflect movement of nuclear material from the nucleolar region into the cytoplasm. The "blebs" are thought to be homologous to structures described by many light microscopists, but they are considerably larger than the nuclear "blebs" described previously by electron microscopists.     

An Electron Microscope Study of Myelin Figures

In the electron microscope, thin sections of OsO₄-fixed myelin figures from the phospholipide fraction of human brain show a pattern of parallel dark lines with a repeating period of about 40 A. It is shown that the dark lines probably represent the reaction product of OsO₄ with double bonds in the fatty acid chains, thereby marking the central portion of one bimolecular lamella. The addition of globin results in dense lines 25 to 50 A wide that cover the surface of the myelin figures. When such a figure consists of only two bimolecular leaflets of lipide covered with globin, the structure shows striking similarity to the image of cell membranes in fixed tissue sections. A hypothetical schema is given of the molecular structure of the figure, and the distribution of OsO₄ in it.     

Scanning Electron Microscope Study of Neisseria gonorrhoeae

Morphological studies utilizing various microscopy techniques have aided in our understanding of the gonococcus and gonorrhea. In this study scanning electron microscopy was used to study differences in virulent (colony types 1 and 2) and avirulent (colony types 3 and 4) gonococci relative to colony appearance, patterns of growth in liquid media, and surface features of individual cocci. Colony types of virulent gonococci are smaller in diameter but have a higher evaluation than those of avirulent mutants. Colony type 2 has a convex undersurface that is associated with surface pitting of solid media. When the colonies are grown in liquid media, various degrees of autoagglutination are observed; this is most pronounced with type 2 and least evident with type 4. Although pili may be involved in this phenomena, other mechanisms must be employed, since type 3 gonococci that lack pili autoagglutinate. Pili are seen on types 1 and 2 and are absent from types 3 and 4. They appear as individual threads radiating from the bacteria or as bundles of pili attaching adjacent cocci. Another extracellular structure consists of small spherical bodies that can coat the bacteria surface, attach to pili, or exist free from other bacterial components. These spheres are least evident with type 4. The gonococcal surface is pebbly with multiple sulci.     

An Electron Microscope Study of the Spermatozoid of Sphagnum

The micro-anatomy of the cilia of Sphagnum has been elucidatedin greater detail than in the ciliary types previously investigated.The facts are summarized in a diagrammatic reconstruction.     

An Electron Microscope Study of Cysts in the Saprolegniaceae

The structure of the zoospore cysts of various members of theSaprolegniaceae has been studied by electron microscopy. InSaprolegnia ferax, S. dioica, in Isoachlya eccentrica and I.unispora the primary cysts were smooth, whilst the secondarycysts bore stalked double-headed hooks. In S. parasitica thesecondary cysts bore tufts of longer hooks. In Protoachlya,Achlya, and Brevilegnia neither type of cyst bore hooks. InDictyuchus sterile the secondary cysts bore large spiny projections.The primary cysts in many species bore tufts of radiating hairs,and it is suggested that these are the remains of ciliated flagella.     

Electron Microscope Study of Ribonucleic Acid of Myxoviruses

Intact ribonucleic acid (RNA) molecules in an extended form were extracted from purified influenza virus and observed in the electron microscope. For this study, the RNA extraction procedure and the Kleinschmidt protein monolayer technique were modified. The mean lengths of RNA from X7, X7-F1, and WSN strains of influenza virus were found to be 2.69, 2.55, and 2.37 mum, respectively. From these measurements, the corresponding estimated molecular weights would be 2.9, 2.8, and 2.5 x 10(6) daltons. X7 and WSN RNA preparations were exposed to pH 3 to disrupt intact molecules. Histograms of length measurements showed five peaks, which were interpreted to represent the five pieces of RNA reported to exist in the influenza virion. X7 RNA appeared to be more stable than WSN RNA when stored at 4 C. The profiles of histograms of incomplete virus RNA suggest that the high molecular-weight component is missing. In preliminary experiments on Newcastle disease virus RNA, molecules of various lengths were observed.     

Studies on Cartilage: Electron Microscope Observations on Normal Rabbit Ear Cartilage

Normal rabbit ear cartilage studied with the light and electron microscope shows chondrocytes in which large lipide spherules, and an abundance of glycogen, a few small mitochondria, and relatively few elements of the endoplasmic reticulum can be identified. The chondrocytes contain, in addition, a material which stains strongly with acid fuchsin and appears in the electron microscope as a relatively dense felt-work. In electron micrographs, the matrix of normal rabbit ear cartilage consists of two components: a uniformly distributed moderately dense substance which appears as a fine meshwork without any particular pattern extending from cartilage cell border to cartilage cell border; and a three-dimensional anastomotic network of more dense material, which is best described as "felt-like" lying between the cells. The similarity between the felt-like material of the matrix and the elastic fibers described in previous electron microscope observations is discussed.     

Structure of the Rete Mirabile in the Kidney of the Rat as Seen with the Electron Microscope

Electron micrographs of the rete mirabile in the medulla of the rat have revealed that the endothelium of the afferent and efferent vessels are markedly different in fine structure. The venous capillaries returning blood from the papilla are lined with a fenestrated endothelium much like that in the peritubular capillaries of the kidney. The arterial capillaries delivering blood to the papilla have an unperforated lining of overlapping endothelial cells with extremely irregular tapered margins. It is pointed out that the organization of particularly the latter vessels suggests that the functional capabilities of these retia go beyond those of a simple diffusion countercurrent exchanger.