An immunohistochemical and electron microscopic study of extra-renal basement membranes in dogs with Samoyed hereditary glomerulopathy

Summary Samoyed hereditary glomerulopathy (SHG) in dogs has been employed as a model for human hereditary nephritis (HN), since affected dogs and patients show splitting of glomerular capillary basement membranes by electron microscopy (EM) and absent staining of glomerular capillaries for Goodpasture antigen (GPA) by immunofluorescence (IF). EM and IF were used to examine basement membranes (BM) in skin, lung, choroid plexus, lens, retina, and inner ear in SHG. By EM, BM in these tissues appeared similar in affected male, carrier female, and unaffected dogs. By IF, GPA could be detected only in lens capsule, internal limiting membrane of retina and basilar membrane of inner ear of unaffected and carrier female dogs, but not in affected male dogs. However, eye abnormalities and hearing loss were not present in any dogs, in contrast to their frequent occurrence in human HN. Our findings on extra-renal BM in SHG suggest that GPA is not required to maintain normal vision or hearing in affected male dogs and permit a greater understanding of the pathogenesis of human HN.The work described in this paper was submitted as partial fulfilment for the Ph.D. degree to Dr. P. Thorner in the Department of Pathology, University of Toronto, Canada     

An electron microscopic study of periderm cell development in mouse limb buds

Summary Development of periderm cells covering fore-and hindlimb buds of mouse em`ryos was observed by scanning and transmission electron microscopy at half day intervals from day 9.5 to 12.5 of gestation (vaginal plug=day 0).At day 9.5, the epidermis is single layered. Occasional periderm cells are present at day 10.5. By day 11.5 a complete layer of periderm cells has covered the entire limb bud.By scanning electron microscopic observation, periderm cells covering the apical ectodermal ridge (AER) are characterized by a small surface size and an elongated polygonal shape with the long axis parallel to the antero-posterior contour of the apical rim. Periderm cells covering the dorsal and ventral surfaces of the limb bud are relatively large and have a polygonal surface shape.The periderm covering the apical tip reflects well the developmental state of the AER. Hence, it is possible to estimate the development of the AER by observing the surface features of the apical periderm by scanning electron microscopy.This work was supported by Scientific research Grant No. 348082 from the Ministry of Education, Japan     

An electron microscopic study of dendritic degeneration in the cerebral cortex resulting from laminar lesions

Summary The morphological characteristics of dendrites in layers of the cerebral cortex above laminar lesions induced by ionizing particle irradiation have been studied in the striate field of rat at various survival times. Within two weeks following irradiation an increasing number of dendrites display unusual alterations inferred to be signs of degeneration.Degenerating dendrites can be characterized by a dense cytoplasmic matrix, disruption of mitochondria, presence of dense bodies, irregular outline and a marked alteration of the plasmalemma in its dimensions and staining properties. Some degenerating dendrites possess a large accumulation of dense subsynaptic material and are contacted by synapses with enlarged and altered synaptic clefts. A few dendrites contain extensive membranous whorls. Engulfment by reactive astrocyte processes is a common feature and often includes the presynaptic axonal knob, but only the degenerating dendrite has been observed within glial cytoplasm. The inference that the majority of degenerating dendrites in this material are apical dendrites of pyramidal cells suggests that either shaft synapses are common for these cells, protuberances may retract during degeneration, or spines are lost due to loss of afferent terminals.Supported by research grants from the United States Public Health Service, NB-4578 and NB-6594.     

Muscle regeneration after exercise-induced myoglobinuria: An electron microscopic study

Muscle regeneration was studied by light and electron microscopy in a case of exercise-induced acute myoglobinuria in a young patient with carnitinepalmityl-transferase deficiency. Various stages of regeneration existed in the foci of necrosis scattered throughout apparently normal muscle. Activated satellite cells, myoblasts and myotubes were found, some of them containing myofibrils. Among the cells accumulating in the necrotic fibres, some apparently contained surviving myonuclei. In some fibres of normal size, developing myofibrils were abundant. Surviving myonuclei may be of significance in the reaction of muscle cells after injury.     

Intrahypothalamic connections: An electron microscopic study in the rat

Summary Terminal degeneration within the hypothalamus was studied by electron microscopy 1 or 2 days (1) after carefully placed microlesions in the arcuate, anterior periventricular, ventromedial, premammillary and posterior hypothalamic nuclei and (2) after microlesions placed in the hypothalamus deafferented 3 weeks earlier.In the median eminence terminal degeneration was found after each of these lesions. Projections from the ventromedial nucleus reach the arcuate, suprachiasmatic, and anterior periventricular nuclei.Projections from the arcuate nucleus terminate in the medial preoptic, anterior periventricular, and ventromedial nuclei.After lesioning the premammillary nuclei degeneration was found in the supraoptic, arcuate, anterior hypothalamic and ventromedial nuclei.     

An electron microscopic radioautographic study on the DNA synthesis of cecal epithelial cells in aging mice

The ultrastructural difference between labeled and unlabeled cecal epithelial cells with tritiated thymidine of prenatal and postnatal developing mice was analyzed by electron microscopic radioautography. The labeled cells were mainly located in the bases of the crypts; silver grains were observed primarily over the nuclei though a few of them were over the mitochondria. The labeling index (L.I.) of cecal epithelial cells was at the highest level at prenatal day 19, decreasing significantly on postnatal day 1, and then having no significant change to 12 postnatal months. Cell organelles were not rich in the cytoplasm of labeled cells. This study was presented at the 26th Annual Meeting of the Clinical Electron Microscopy Society of Japan, Kochi, October 6, 1994.     

An electron microscopic study of the pars distalis of the dog adenohypophysis

Summary The parenchymal cells of the pars distalis of the dog adenohypophysis were studied with the electron microscope. Criteria for the differentiation of four cell types included the size and shape of the cells and the size, shape and distribution of organelles and secretory granules. Three secretory granule-containing cells designated as I, II and III were observed. Cell type I was further divided into subtypes (Ia, Ib, Ic) on the basis of the granule size of maximal frequency. A fourth cell type (IV) contained no or only a few secretory granules. It was unique in having a highly irregular shape with cytoplasmic processes extending between the other cell types and also occasionally enclosing follicular cavities.These studies were supported by USPHS training grant, GM-155 and by Public Health Service, GRS grant 5394. The author wishes to thank Dr. J. M. Wolfe and Dr. E. V. Crabill for their valuable assistance during the course of this investigation.     

Light and electron microscopic studies of collagen fibers under strain

Summary The correlation of the mechanical properties and the morphological behaviour of collagenous tissue during strain was investigated. Relaxed and strained tendons were examined by incidental and transmitted polarized light microscopy and electron microscopy. It was concluded that the wavy pattern seen at the seminicroscopic level of the collagen fibers vanished at relatively low loads. The direction of birefringence in transmitted light microscopy varied in the relaxed preparations but became even in the strained. The electron microscopy showed that the fibrils had the same periodicity as calculated from X-ray diffraction patterns of moist collagen, i.e. 680 Å, and that straining increased the period lengths of some fibrils. The correlation of these findings to a mechanical analogy of the tissue was discussed.     

A study by immunofluorescence microscopy of the NC1 domain of collagen type IV in glomerular basement membranes of two patients with hereditary nephritis

Summary The NC1 domain of the collagen type IV molecule, the major component of glomerular basement membranes (GBM), consists of dimers and 24 kilodalton (K), 26 K and 28 K monomers in man, and contains the Goodpasture antigen. Serum obtained from patients with Goodpasture's syndrome has been reported not to stain GBM of most male and some female patients with hereditary nephritis (HN) by immunofluorescence (IF) microscopy. In the present study, GBM seen on the renal biopsies of 2 patients (one male and one female) with HN were examined by IF to ascertain whether NC1 monomers were detectable. Three reagents were used: a plasmapheresis fluid (PPF) obtained from a patient who was treated for anti-GBM nephritis (human anti-GBM PPF); a commercial rabbit antibody against human NC1; and a rabbit antibody raised by us against dog NC1, which cross-reacted with human NC1. All 3 reagents detected NC1 determinants in GBM of normal human kidney by IF and reacted with human NC1 by a plate-binding radioimmunoassay (RIA). The human anti-GBM PPF bound to 28 K and 26 K monomer components of NC1 by Western blotting, the rabbit anti-human NC1 antibody bound to 26 K and 24 K monomers, while the rabbit anti-dog NC1 antibody bound only to the 26 K monomer. By IF, the human anti-GBM PPF did not stain GBM of the male patient with HN, but produced segmental staining of GBM (i.e., some GBM stained, while others did not) of the female patient. In contrast, the rabbit anti-NC1 antibodies produced global staining by IF of GBM of both patients. The absence of staining (i.e., global or segmental) seen with the human anti-GBM PPF implied that the 26 K and 28 K monomers of NC1 were either absent from GBM, or were present but altered structurally, leading to a diminution in their immunological reactivity. However, the positive staining observed with the rabbit anti-NC1 antibodies implied that the 26 K monomer was actually present in GBM. Hence, we postulate that the 26 K monomer of NC1 in GBM was structurally altered, and that the 28 K monomer was either absent, or present but altered. These findings suggest that there is an abnormality of more than one monomer of NC1 in GBM of patients with HN.     

Chronic megakaryocytic-granulocytic myelosis — An electron microscopic study

Summary The fine structure of the bone marrow in chronic megakaryocytic-granulocytic myelosis (CMGM) was studied in 5 nontreated patients to investigate possible malignant proliferation of megakaryocytes and the role of megakaryopoiesis in fibrillogenesis, terminating in osteomyelofibrosis. In comparison with normal megakaryopoiesis there is an enormous increase of the megakaryocytic cell line and many immature and atypical forms are seen. Most conspicuous are microforms, nuclear-cytoplasmic disorganization and nuclear inclusions. Asynchrony of maturation causes abnormal thrombocytogenesis with premature detachment of platelets resulting in immature and peculiar giant forms of thrombocytes. Besides megakaryocytes appearing superficially normal the maturation anarchy of many cells is so severe that by analogy with observations in other leukaemic cells these abnormalities are thought to be representative of a malignant growth. Moreover, there is a striking accumulation of microfibrils and single collagen fibres around megakaryoblasts. Since these cells contain all those organelles commonly associated with fibre production the initial step for fibrillogenesis may therefore arise from the megakaryoblasts prior to platelet release, or any fibroblast proliferation.Abbreviations CMGM Chronic megakaryocytic-granulocytic myelosis - CGL Chronic granulocytic leukaemia - MF Myelofibrosis - OMS Osteomyelosclerosis - DMS Demarcation system - RER Rough surfaced endoplasmic reticulum Supported by the Deutsche Forschungsgemeinschaft, grant Ge 121/15     

An electron microscopic study of plasmatic arterionecrosis in the human cerebral arteries

Summary An electron microscopic study of the intracerebral arteries from 9 hypertensive cases was performed in order to elucidate the morphogenesis of the plasmatic arterionecrosis which was considered to be the direct cause of hypertensive intracerebral hemorrhage. In the preceding stage of the arterial lesions, marked necrosis of medial smooth muscle cells and increase of basement membrane-like substance in the intima and media were observed. The lumina of these arteries were slightly dilated. The dilatation and hemodynamic factors were supposed to cause endothelial injury resulting in blood plasma insudation into the intima through the opened spaces between endothelial cells. The insudated blood plasma dispersed and dissolved the basement membrane-like substance, collagen and elastic fibers in the arterial wall, leading to the development of the plasmatic arterionecrosis.This study was supported by a Grant-in-Aid for Scientific Research of the Japanese Ministry of Education (No. 857052).     

Quail Rathke''s pouch differentiation an electron microscopic study

Summary Adenohypophyseal region of quail embryo has been examined by electron microscopy from stage 12 to stage 21 of Zacchei (1961).The Seessel's pouch develops prior to the early stages of adenohypophysis formation, then regresses while Rathke's pouch proliferates and differentiates.From Rathke's pouch formation by stage 12 (48 h of incubation) until appearance of the first secretory granules by stage 21 (6 days of incubation), there are no major ultrastructural modifications in adenohypophyseal cells. Mitochondria, Golgi vesicles, polysomic ribosomes, pinocytotic vesicles, and mitotic figures become more numerous while nucleocytoplasmic ratio and the number of ribosomes and lipid droplets decreases. The major change is the appearance of secretory granules by day 6 of incubation. This phenomenon occurs at the same time as in chick embryo, despite an incubation period shorter for quail than for chick. Mitotic figures are mainly distributed near the pouch lumen, while secretory granules are first located in the peripheral cells of the cephalic part ofpars distalis primordium. The hypothetical role of mesenchyme and vascularization is discussed.This work has been supported by a grant from DGRST, no. 77.7.9665     

An electron microscopic and immunohistochemical study of Merkel cell carcinoma

Two cases of Merkel cell carcinoma (MCC) were examined by electron microscopy and immunohistochemistry. Histologically, tumor cells, which extended from the dermis into the subcutis, showed anastomosing bands with partial trabecular pattern. The ultrastructural study showed tumor cells in case 1 with numerous neurosecretory granules. The number of granules in case 2, however, was smaller compared with that in case 1. Perinuclear bundles of filaments were present in case 2, but few bundles were observed in case 1. By immunohistochemistry, cytokeratin (CK)-8, -18, -19, and -20 and epithelial membrane antigen were stained positively within tumor cells in both cases. It was interesting that staining patterns of chromogranin A and of neuron-specific enolase were different in the two cases. These data indicated that CK-20 is a useful marker for diagnosing MCC and that ultrastructural and immunohistochemical differences in both cases were the result of phenotypic variation.     

An electron microscopic study of the developing caudate nucleus in euthyroid and hypothyroid states

Summary Thyroid hormone exerts a powerful influence on CNS growth and maturation. Hypothyroidism early in life has long been known to cause disturbances in innate behavior, motor performance, severe and frequently irreversible mental retardation. In this deficiency, depressed caudate neurogenesis, cell migration and neuropil development during the rapid period of CNS growth may contribute to the clinical picture of perceptual handicaps often seen in cretins. Light microscopic and Golgi studies of the developing caudate nucleus in thyroid deficiency have been carried out to help attain insights into the mechanisms whereby the extrapyramidal system regulates motor function. The ultrastructural study of caudate nuclear cytogenesis and synaptogenesis in normal and hypothyroid states provides more detailed information for further analysis of the problem.Hypothyroidism was induced from birth by adding propylthiouracil to the food and drinking water of lactating dams. Linear development of the caudate nucleus of both normal and hypothyroid rats at ages 8, 14, 20, 30 and 42 days was studied by electron microscopy. Thyroid glands were examined by light microscopy to assess the normal and deficient states.Immature cells, primitive processes and synapses were the characteristic features of the 8-day-old normal caudate nucleus. Distinctively wide cisternae of the rough endoplasmic reticulum, loosely packed Golgi apparatus and chromatin clumps throughout the nuclei of the neurons were significant early morphologic variations. The dramatic cytoarchitectural maturation in the 14- and 20-day normal caudate neuropil points to the rapidity of developmental rate. After the growth spurt of the first three weeks a maturational plateau occurs which is characterized by well-formed neuronal cytoplasmic organelles, myelinated and non-myelinated axons, axon terminals, dendrites and their spines, and synapses.Thyroid deficiency causes a marked maturational delay of approximately 7 days in caudate neuronal proliferation, the elaboration of neuronal networks and the attainment of mature synaptic contents and membranes. This delay is evidenced by comparison of the structural similarities between 8-day-old normal and 14-day-old deficient rats; and additional comparisons between the 14-day-old normal and 20-day-old hypothyroid rats. A rapid catch up process in fine structural morphogenesis takes place in the period between days 14 and 30 in the deficient animals. Repression of thyroid function does not entirely prevent development of the caudate nucleus but allows a fairly extensive, though critically incomplete degree of maturation. This imperfection is manifested by a decrease in the number of synaptic contacts that persists even after the rapid catch up phenomenon of caudate synaptogenesis.Partially supported by USPHS Grant No. HD-05615     

An electron microscopic study of neurosecretion in the cerebral ganglion of the earthworm

Non-synaptic, exocytotic release of neurosecretory granules in cerebral ganglion neurons was observed electron microscopically in 3 species of the oligochaete annelids Aporrectodea caliginosa, Octolasion cyaneum and Lumbricus terrestris. In addition to the features indicating exocytotic release of neurosecretory granules into perineuronal space, possible features of neurosecretion into blood vessels were seen within the cerebral ganglion. Axon terminals in synaptic contact with perikaryal profiles of cerebral ganglion neurons were also found.     

Chronic megakaryocytic-granulocytic myelosis — An electron microscopic study including freeze-fracture

Summary In 5 patients with chronic megakaryocytic-granulocytic myelosis (CMGM) bone marrow specimens were studied by electron microscopy to investigate possible abnormalities of the granulocytic cell lineage. Thin sections were compared with freeze-fracture replicas to elucidate further aspects of leucocyte cytology. The atypia exhibited in these cells (eosinophils, basophils and neutrophil granulocytes) consisted mostly of a disorganization of granulopoiesis with hyper- and hypogranulation, a pathological increase in the number of nuclear blebs and a maturation asynchrony sometimes leading to Pelger-like cell forms. Moreover, a presumptive stem cell was demonstrated in the erythopoietic and granulocytic cell lines resembling CFU cells. In conclusion, granulopoiesis in CMGM exhibited abnormalities as generally observed in chronic myelogenous leukaemia. When considered with our previous finding of malignancy in megakaryopoiesis, CMGM has to be classified as a myelosis of mixed cellularity.Abbreviations CMGM Chronic megakaryocytic-granulocytic myelosis - CGL Chronic granulocytic leukaemia - MF Osteomyelofibrosis - RER Rough surfaced endoplasmic reticulum Supported by the Deutsche Forschungsgemeinschaft, grant Ge 121/15     

An electron microscopic study of varicella skin lesions

Summary The skin lesions of varicella were studied by electron microscopy. Singlemembrane particles were the only type of viral particles present in the nucleus. Two modes of viral envelopment were observed; budding from the inner nuclear membrane, and budding from vacuole membranes of the cytoplasm. Tubular, filamentous structures, probably an aberrant form of the virus, were often observed in affected nuclei. Nuclear bodies were frequently found and showed abnormal structures in the nuclei of cells near the intraepidermal vesicles. Strands, inclusion-like structures, probably composed of degenerated nuclear materials, were seen in some instances in the cytoplasm. Virions were frequently included in phagosomes of epidermal or dermal macrophages.     

Cytoarchitecture of the substantia nigra pars lateralis in the opossum (Didelphis virginiana): A correlated light and electron microscopic study

Background: The substantia nigra has been divided into three subdivisions. However, the cytoarchitecture of one of these subdivisions, the pars lateralis (SNI), has not been previously examined in detail at the light and electron microscopic levels in any species. In the adult opossum, the three nigral subdivisions can be easily distinguished as distinct, rostrocaudally oriented cell groups separated by neuron-free zones. Thus it was possible to determine the boundaries of the SNI unambiguously. This report covers the results of an examination of the morphology and organization of the SNI in the opossum. Methods: Material from 13 opossums was used for this study. Eight of the animals had been previously stained for Nissl substance (n=4) or impregnated by the Golgi technique (n=4). The remaining five animals were prepared for electron microscopic studies using standard procedures. Results: Two cell types were identified on the basis of morphological differences, small and medium-large neurons. Small neurons (10–18 μm long axis) have large nuclei with moderate amounts of heterochromatin and a thin rim of cytoplasm. They have long (up to 500 μm), spine-free dendrites. Medium-large neurons (18–54 μm long axis) have rounded nuclei with electron-lucent nucleoplasm. Few indentations of the nuclear envelope were observed. The surrounding cytoplasm has dense arrays of organelles. Nissl bodies are particularly prominent in the form of pyramids with their bases at juxtanuclear positions and their apices directed toward emerging dendrites. Dendrites of medium-large neurons are long (some>1 mm in length), are primarily oriented in the frontal plane, and extend along the dorsal surface of or into the cerebral peduncle. Some cells have dendrites that are moderately spinous, whereas other neurons possess sparsely spinous dendrites. Relatively few synaptic profiles are observed to contact somata and proximal dendrites. Conclusion: This report provides added morphological support for the idea that the SNI is a distinct subdivision of the substantia nigra, a distinction previously made on the basis of the physiologically characterized relationship between the lateral substantia nigra and orienting behaviors and seizure-related function. © 1995 Wiley-Liss, Inc.     

Primary linitis plastica type carcinoma of the small intestine: An immunohistochemical and electron microscopic study

We examined a case of an 87-year-old man who had had a 1-month history of ileus of unknown cause. Postmortem examination revealed numerous foci of constriction and thickening throughout the small intestine, associated with perforation and panperitonitis. Histologically. diffuse infiltration of anaplastic atypical cells was found in the wall of almost the entire small intestine. The atypical cells were immunohistochemically positive for carcinoembryonic antigen and epithelial membrane antigen. In the electron microscopic examination, most of those cells had microvilli with filamentous core rootlets and terminal webs. There was no atypical cell infiltration in the stomach or large intestine. We concluded that this case was primary linitis plastica type carcinoma of the small intestine involving almost the entire small intestine.     

Standard methods of electron microscopic radioautography

The standard methods of making electron microscopic radioautograms and quantitation of silver grains in our laboratory were presented. Specialized techniques for insoluble or soluble compounds should be employed for demonstrating the synthesis of various macromolecules and the incorporation of various labeled substances. Specimens for electron microscopic radioautography processed under the same conditions can be employed for the quantitation of radioactive substances in subcellular compartments using image analyzers.