A new type of inclusion body in human spinal cord

Summary New pericapillary inclusion bodies were found in 17 cases of sporadic amyotrophic lateral sclerosis (ALS). The inclusion bodies consisted of paracrystalline arrays with 5–7 nm electron-dense subunits, were discernible with the light microscope and had the staining properties of protein. They were surrounded by capillary-wall basement membrane and were often associated with peripheral fibrils. Astrocytic fibrillary bodies, without paracrystalline material, were also found. The ultrastructure and staining of the fibrils suggests that the paracrystalline material is within pericapillary astrocytes. The nature and significance of the inclusion bodies are unknown, but their presence suggests that there may be pericapillary abnormalities in the spinal cord in ALS and possibly other disorders.Supported by the Montreal General Hospital Research Institute     

Hirano bodies contain tau protein

Hirano bodies are intraneuronal inclusions whose frequency increases with age and Alzheimer's disease. These paracrystalline inclusions have been shown previously using immunocytochemistry to share epitopes with actin, tropomyosin, alpha-actinin and vinculin. Hirano bodies have not previously been demonstrated to share components with neurofibrillary tangles, another intraneuronal inclusion characteristic of Alzheimer's disease. In this study, we show that Hirano bodies bind antibodies to the microtubule-associated protein tau, a component of Alzheimer neurofibrillary tangles.     

Mitochondrial myopathy in Marinesco-Sjögren syndrome

ABSTRACT. Myopathy represents one of the major features of Marinesco-Sjögren syndrome (MSS). Seven patients with MSS from three different families were studied with morphological and neurophysiological methods. In two patients ragged-red fibres were found after Gomori trichrome staining. Transmission electron microscopy (EM) showed that subsarcolemmal accumulation of abnormal mitochondria regularly occurred and in one patient paracrystalline inclusion bodies were found. The EMG showed myopathic changes while the nerve condition velocities were normal. A delayed normalization of exercised-induced hyperlactatemia was noted. These findings show that mitochondrial myopathic changes are present in MSS.     

A clinicopathological study of neuronal ceroid lipofuscinosis]

4 unrelated patients with infantile type (one case), late infantile type (one case), juvenile type (two case) of neuronal ceroid-lipofuscinosis were reported. The pathological study by brain biopsy was carried out with light and electron microscopes as well as histochemical methods. Ultrastructurally, the infantile type showed deposits of lipofuscin bodies, fingerprint bodies, and the late infantile type showed a lot of curvilinear bodies and paracrystalline bodies. The ultrastructural configuration in the juvenile type is more heterogeneous and consisted of combination of lipofuscin bodies, fingerprint bodies and multilamellar structures. It was emphasized that fingerprint bodies were first discovered in infantile type, which might challenge the view that only lipofuscin bodies can be found in infantile type.     

Tissue mosaicism in the skeletal muscle and sural nerve biopsies in the MELAS syndrome

We describe a clinically full-blown MELAS patient, who had an A3243G point mutation of mitochondrial DNA (mtDNA) in muscle and blood cells, and his family members. From the proband two muscle biopsies from the vastus lateralis muscle were analysed; one had typical ragged red fibers and focal cytochrome c oxidase deficiency and the other was completely normal. He also had a peripheral neuropathy confirmed by nerve conduction velocity and sural nerve biopsy studies. Axonal degeneration, relative loss of large myelinated fibers and paracrystalline inclusion bodies in the Schwann cells were noted. Intriguingly, the A3243G mutation of mtDNA was not found in the sural nerve biopsy. Therefore, we conclude that tissue mosaicism is present in the muscle fibers and that the mtDNA mutation may not be detected in the nerve involved as proved by pathology. We also suggest that the involvement of specific tissues in patients with mitochondrial diseases should be further determined by single fiber mtDNA analysis.     

16例线粒体肌病和线粒体脑肌病电镜观察分析

目的探讨线粒体肌病与脑肌病患者肌肉的超微结构特征,分析该病的病因和可能的发病机制。方法对16例线粒体肌病与脑肌病患者的肌活检组织进行光镜和电镜超微病理观察。结果电镜观察16例,在肌原纤维间和肌膜下可见弥漫的线粒体数目增多,13例表现为形态异常,可见巨大线粒体,嵴结构不清,排列紊乱,呈同心圆样,均可见线粒体内类结晶状包涵体,有的同时伴有糖原颗粒的异常增多、脂滴沉积及溶酶体异常,有的线粒体只能靠双层膜结构及残存的嵴被识别。3例仅线粒体数量增多,未见其他异常。结论电镜观察肌膜下和肌原纤维间线粒体异常增多且形态异常,特别是线粒体内类结晶状包涵体,对本病的诊断有重要价值。     

Lactacystin诱导PC12细胞胞内嗜酸性包涵体动态变化的研究

目的 研究蛋白酶体抑制剂lactacystin诱导PC12细胞胞浆内嗜酸性包涵体的动态变化过程.方法 PC12细胞中加入不同浓度lactacystin(0、5、10、20μmol/L),孵育24 h后行HE染色及α-synuclein免疫组织化学染色,光镜观察包涵体变化;其中10 μmol/L lactacystin组分别于加入lactacystin后24、36、48、72、96和120 h行上述染色法,光镜观察包涵体变化.结果 lacatacystin作用于PC12细胞24 h后,HE染色显示随着浓度增加.胞浆中嗜伊红包涵体数量逐渐增多.0 μmol/L组胞浆中未看到包涵体,5 μmol/L组胞浆中有少量包涵体出现(3.33%±1.15%),10μmol/L组多数细胞胞浆出现典型球状包涵体(71.33%±4.16%),20μmol/L组几乎每个细胞胞浆中均出现嗜伊红包涵体(90.33%±3.21%),个别包涵体游离于细胞外;10 μmol/L组随着时间延长,包涵体逐渐从胞浆中游离至细胞外,其后胞浆逐渐缺失,在96 h和120 h时仅剩余胞核和球状包涵体.免疫组化染色显示:lactacystin作用24 h时,胞浆中α-synuclein免疫反应阳性物质逐渐由散在颗粒逐渐聚集成球状包涵体;随着浓度的增加和时间延长,α-synuclein染色阳性的球状包涵体游走于细胞外,胞浆逐渐缺失,至96 h和120 h时仅剩余胞核和包涵体.结论 lactacystin作用于PC12细胞后,早期在胞浆中出现散在的嗜酸性和α-synuclein免疫反应阳性的颗粒,随着浓度增加和时间延长,颗粒状物质在核旁聚集,最终形成固缩的包涵体,细胞外包涵体可独立存在.这种包涵体的动态变化过程与以人类帕金森病为代表的神经变性疾病中的包涵体非常相似,可以作为研究包涵体相关问题的有力工具.     

High molecular weight microtubule-associated proteins bind to actin lattices (Hirano bodies)

Summary Hirano bodies are filamentous, paracrystalline inclusions that are found in dendrites and cell bodies of neurons in Alzheimer's and other neurodegenerative diseases. Actin appears to be a major component of these structures. We present evidence that tropomyosin and high molecular weight microtubule-associated proteins (MAPs) are also components of Hirano bodies. Although an association betwen actin and MAPs has been noted in vitro, interactions in vivo have not heretofore been demonstrated. Since microtubules are not present in Hirano bodies, and anti-tubulin and anti-neurofilament antibodies do not bind to Hirano bodies, the association between MAPs and these inclusions is likely a result of interactions between MAPs and actin, and not MAPs and microtubules or neurofilaments.Supported in part by grants AG05386, The Alzheimer's Disease and Related Disorders Association, The John Douglas French Foundation, and The Wolf Memorial Fund     

Lymphocyte ultrastructure in two cases of neuronal ceroid-lipofuscinosis.

Circulating blood lymphocytes from two patients with neuronal ceroid-lipofuscinosis (NCL) were investigated by transmission electronmicroscopy. Ultrastructural examination showed two forms of intracytoplasmic single membrane-limited inclusions. Contents of the first inclusion form were arranged in five distinct patterns: (1) granules, (2) membranous formations, (3) paracrystalline forms, (4) alternating electron-dense/electron-lucent arrangements, and (5) admixtures of these components. These molecular morphologies suggest the usefulness of lymphocyte fine structure as a diagnostic tool in NCL. The second inclusion form contained cylinder-like structures. These structures are not specific for NCL and have been identified in other diseases.     

Mitochondrial changes in acute myopathy after treatment of respiratory failure with mechanical ventilation (acute relaxant-steroid myopathy)

A case of acute myopathy was observed in the course of treatment of respiratory failure with mechanical ventilation combined with prolonged neuromuscular blockade and administration of corticosteroids. A muscle biopsy revealed degeneration of muscle fibres. Electron microscopy showed loss of thick filaments as well as nemaline rods, vacuoles and cytoplasmic bodies. The mitochondria were increased in number, many harbouring paracrystalline inclusions, which were hitherto unknown in this condition.     

The significance of ragged-red fibres in neuromuscular disease

The pathological significance of ragged-red fibres is uncertain. We have studied ragged-red fibres in the muscle biopsies of 3 adults; one with polymyositis and two with progressive external ophthalmoplegia. All the ragged-red fibres were Type 1 fibres. In two patients the mean diameter of the ragged-red fibres was significantly smaller than the unaffected Type 1 fibres. Some of these fibres showed features of regeneration, and others of degeneration. In the patient with polymyositis the mitochondria were proliferated and contained osmiophilic dense bodies; in the other two patients paracrystalline mitochondrial inclusions were prominent. These findings suggest that ragged-red fibres do not represent a single pathological process.     

A cell culture model for investigation of Hirano bodies

Hirano bodies are paracrystalline F-actin-rich aggregations associated with a variety of conditions including aging, and neurodegenerative diseases. The composition and structure of these inclusions have been described by immunohistochemistry and ultrastructure, respectively. However, studies of the physiological function and dynamics of Hirano bodies have been hindered due to lack of a facile in vitro experimental system. We have developed a model for formation of Hirano bodies in mammalian cell cultures by expression of the carboxy-terminal fragment (CT) of a 34-kDa actin-bundling protein. Expression of the CT protein induces F-actin rearrangement in HEK 293, HeLa, Cos7 cells, neuroblastoma and astrocytic cells, and in primary neurons. We have termed these structures model Hirano bodies, since their composition and ultrastructure is quite similar to that reported in vivo. Model Hirano bodies in cell cultures sometimes appeared to be formed of a number of smaller domains, suggesting that small aggregates are intermediates in the formation of Hirano bodies. Stable lines expressing CT and bearing model Hirano bodies exhibit normal growth, morphology, and motility. This model provides a valuable system for the study of the dynamics of Hirano bodies, and their role in disease processes.     

Parkinson's disease and dementia with neuronal inclusions in the cerebral cortex: Lewy bodies or Pick bodies

Cortical Lewy bodies may be difficult to differentiate from Pick bodies by the usual staining methods. This problem is illustrated in a case of progressive dementia with parkinsonian features. Lewy bodies were found, not only in the pigmented nuclei in the brainstem and in the cerebral cortex, but also in the dentate fascia, a predilection site for Pick bodies. The inclusion bodies were compared with the inclusion bodies in a case of Pick's disease. Intense argyrophilia of the cortical inclusion bodies argued in favor of Pick bodies; antibodies to phosphorylated neurofilaments reacted with the cortical inclusions and with classical Pick bodies, but antibodies to paired helical filaments reacted only with the Pick bodies. The most convincing evidence, that the inclusions were Lewy bodies, was obtained by electron microscopy. The filaments in the inclusions showed fuzzy deposits of electron dense material, characteristic for filaments of Lewy bodies. This contrasted with the smooth filaments of the Pick bodies. It is concluded that cortical inclusions in brains from patients with Lewy bodies in the pigmented nuclei of the brainstem most likely represent Lewy bodies. This can be confirmed by examining the ultrastructure of the inclusions.     

Dying-back oligodendrogliopathy: A late sequel of myelin-associated glycoprotein deficiency

Ultrastructural analysis of myelin from 8-month-old mice deficient in the myelin-associated glycoprotein revealed pronounced and characteristic alterations of the periaxonal oligodendrocyte processes, consisting of intracytoplasmic deposition of vesicular material, multivesicular bodies, mitochondria, and lipofuscin granules, as well as granular or paracrystalline inclusions. These alterations are similar to those described before as “dying-back oligodendrogliopathy” in diseases of toxic or immune-mediated demyelination including multiple sclerosis. © 1997 Wiley-Liss Inc.     

Neuropathological changes of the brain in myotonic dystrophy--some new observations

Brain autopsy materials from 2 patients with myotonic dystrophy (MyD) were studied. The results obtained in these 2 cases were quite similar. Besides thalamic inclusion bodies and minor abnormalities in gyral architecture with a disordered cortical cellular arrangement, some new observations have been made. First, no more than one intracytoplasmic inclusion body per cell was present in the cerebral cortex, the thalamus, the caudate nucleus and the putamen; this inclusion body was oval or elongated with smooth, sharply defined contours and was usually located at the periphery of the cell. Second, irregular intracytoplasmic inclusion bodies, often multiple and not surrounded by a halo, were found at the periphery or within accumulations of neuromelanin granules in the pigmented cells of the substantia nigra. All the bodies described above stained highly eosinophilic with hematoxylin-eosin and the ultrastructure of the bodies in the thalamus and the substantia nigra was almost the same; these bodies were composed of stacks of alternating parallel, light and dark rectilinear profiles oriented perpendicularly to the longitudinal axis of the bodies. Third, Marinesco bodies were observed with a very high frequency in the pigmented cells of the substantia nigra.     

A case of mitochondrial enzymopathy

A case of mitochondrial enzymopathy, called also ophthalmoplegia plus, was observed in a 31-year-old man. Histoenzymatic investigations demonstrated in the myocytes decreased and irregularity of reactions for succinic dehydrogenase, tetrazole reductase and mitochondrial ATPase. In electron microscopy paracrystalline structures, lamellar bodies and concentrically condensed cristae were seen in the mitochondria, and increased glycogen stores outside the mitochondria.     

Sequestration of Tubulin in Neurons in Alzheimer's disease

In Alzheimer's disease (AD), a variety of populations of neurons exhibits cytoskeletal abnormalities, including neurofibrillary tangles (NFT) in perikarya, Hirano bodies in dendrites and filament-distended axons/terminals/dendrites (neurites) in senile plaques. Some nerve cells, particularly pyramidal neurons of the hippocampus, also develop Hirano bodies (paracrystalline arrays of actin) and granulovacuolar degeneration (GVD; granular inclusions in cytoplasmic vacuoles). Since abnormalities of cytoskeletal elements have been implicated in the formation of NFT, neurites and Hirano bodies, the present study was designed to determine whether GVD also may represent a type of cytoskeletal pathology. Sections of hippocampus from controls and from individuals with AD were stained by immunocytochemical methods using monoclonal and polyclonal antibodies directed against a variety of cytoskeletal antigens. Granules of GVD contained tubulin-like immunoreactivity and absorption with purified tubulin abolished staining. Other antigens were not demonstrated in granules when antibodies directed against other cytoskeletal antigens were used. The observation of sequestration of tubulin in granules is consistent with the concept that abnormalities of the neuronal cytoskeleton are an important part of the cellular pathology of AD.     

Intracytoplasmic inclusion bodies of the thalamus and the substantia nigra,and Marinesco bodies in myotonic dystrophy: a quantitative morphological study

Summary Intracytoplasmic inclusion bodies of the thalamus and the substantia nigra, and Marinesco bodies have been studied in four patients with myotonic dystrophy (MyD), eight patients with other neurological diseases (control A), and eight patients without neurological diseases (control B). The percentages of the affected cells were calculated by dividing the number of neurons including intracytoplasmic inclusion bodies of the thalamus and the substantia nigra, and Marinesco bodies, by the total cell count in these respective regions. Statistical analyses were performed with regard to the frequency of these bodies by using Student'st test. There was a significantly higher incidence of intracytoplasmic inclusion bodies of the thalamus (13.2% versus 0.7%,P<0.001) and the substantia nigra (20.4% versus 2.7%,P<0.001), and Marinesco bodies (37.4% versus 4.1%,P<0.001) in patients with MyD than in controls A and B. From our observations, it is suggested that the presence with a high frequency, in combination, of these bodies is not an incidental finding but may have an intimate and important relationship with the pathogenesis of MyD, and may be a conspicuous and diagnostically important feature of MyD.Supported by Grant-in-Aid for Scientific Research from The Ministry of Education, Science and Culture     

Extrahypophyseal distribution of alpha-melanocyte stimulating hormone (alpha-MSH)-like immunoreactivity in postmortem brains from normal subjects and Alzheimer-type dementia patients

In Alzheimer's disease (AD), a variety of populations of neurons exhibits cytoskeletal abnormalities, including neurofibrillary tangles (NFT) in perikarya, Hirano bodies in dendrites and filament-distended axons/terminals/dendrites (neurites) in senile plaques. Some nerve cells, particularly pyramidal neurons of the hippocampus, also develop Hirano bodies (paracrystalline arrays of actin) and granulovacuolar degeneration (GVD; granular inclusions in cytoplasmic vacuoles). Since abnormalities of cytoskeletal elements have been implicated in the formation of NFT, neurites and Hirano bodies, the present study was designed to determine whether GVD also may represent a type of cytoskeletal pathology. Sections of hippocampus from controls and from individuals with AD were stained by immunocytochemical methods using monoclonal and polyclonal antibodies directed against a variety of cytoskeletal antigens. Granules of GVD contained tubulin-like immunoreactivity and absorption with purified tubulin abolished staining. Other antigens were not demonstrated in granules when antibodies directed against other cytoskeletal antigens were used. The observation of sequestration of tubulin in granules is consistent with the concept that abnormalities of the neuronal cytoskeleton are an important part of the cellular pathology of AD.     

An electron microscopic study of inclusion bodies in synaptic terminals of scrapie-infected animals

Summary Inclusion bodies consisting of vesicles of about 25 nm diameter and occurring in the synaptic terminals of scrapie-infected animals have been described by a number of people. In the present study these inclusion bodies were looked for in the neocortex, hippocampus and corpus callosum in a variety of strains of mice (C3H, LM, RIII, IM, VL) infected with different strains of scrapie agent (22C, 79A, ME7, 87V) after intracerebral inoculation. In plaque-bearing models of scrapie, terminals containing synaptic inclusion bodies were frequently found surrounding the amyloid plaque cores in the neocortex but not in the corpus callosum. In non-plaque-bearing models, terminals containing synaptic inclusion bodies were found in the neuropil of the neocortex and hippocampus. For all models, these bodies were either presynaptic or postsynaptic but were not, as a rule, found on both sides of the same synapse. Fibrillary material was frequently seen in the postsynaptic terminals containing the inclusion bodies in both the plaque- and non-plaque-bearing models. On one occasion fibrillary material was seen, together with the inclusion bodies, in a neuron cell body. Inclusion bodies were also seen in the neocortex of hamsters infected with the 263K strain of scrapie agent and a Cheviot sheep infected with the ME7 strain of agent. The inclusion bodies and the fibrillary material were thought to be derived from the breakdown of neurotubules.