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At least 793 records · Page 44Linked to original sources
PubMed1999

Huntington's disease intranuclear inclusions contain truncated, ubiquitinated huntingtin protein.

Intranuclear inclusion bodies are a shared pathological feature of Huntington's disease (HD) and its transgenic mouse model. Using a panel of antibodies spanning the entire huntingtin molecule, we have investigated the pattern of immunoreactivity within the intranuclear inclusions in the frontal cortex and striatum of patients with HD. The intranuclear inclusions reacted with anti-ubiquitin and antibodies against the N-terminal portion of huntingtin (CAG53b, HP1), but not with HD1 and the 1C2 antibodies that detect the expanded polyglutamine tract nor the more C-terminal antibodies. However, the 1C2, HP1, CAG53b, and HD1 antibodies detected granular cytoplasmic deposits in cortical and striatal neurons that also contained intranuclear N-terminal huntingtin immunoreactivity. These data show a differential intracellular location of truncated huntingtin in the HD brain. Both the cytoplasmic and the nuclear aggregates of the protein fragments could be neurotoxic. The frequency of the cortical intranuclear inclusions correlated with the size of CAG expansion and was inversely related to the age at onset and death. No such correlations were detected for the striatum, which most likely reflects a more advanced neuronal loss accrued by the time of death.

Adult↗
PubMed1993

Overcoming inclusion body formation in a high-level expression system.

Attempts at overexpressing T4-phage deoxycytidylate deaminase using the pET3c/BL21(DE3)/pLysS system resulted in this enzyme being part of an inactive inclusion-body complex. However, by employing an enriched growth medium it was found that the deaminase could be induced in a soluble active form to at least 20% of this organism's cellular protein. Insoluble inclusion bodies were obtained with less rich media. This procedure was employed successfully with other highly expressed proteins that formed inclusion bodies. The use of a rich growth medium during the course of protein induction may be a valuable adjunct to limiting inclusion body formation with this as well as other expression systems.

Bacteriophage T4↗
PubMed2002

Chromatographic methods for the isolation of, and refolding of proteins from, Escherichia coli inclusion bodies.

New methods for the chromatographic isolation of inclusion bodies directly from crude Escherichia coli homogenates and for the refolding of denatured protein are presented. The traditional method of differential centrifugation for the isolation of purified inclusion bodies is replaced by a single gel-filtration step. The principle is that the exclusion limit of the gel particles is chosen such that only the inclusion bodies are excluded, i.e., all other components of the crude homogenate penetrate the gel under the conditions selected. In the novel column refolding process, a decreasing gradient of denaturant (urea or Gu-HCl), combined with an increasing pH gradient, is introduced into a gel-filtration column packed with a gel medium that has an exclusion limit lower than the molecular mass of the protein to be refolded. A limited sample volume of the protein, dissolved in the highest denaturant concentration at the lowest pH of the selected gradient combination, is applied to the column. During the course of elution, the zone of denatured protein moves down the column at a speed approximately threefold higher than that of the denaturant. This means that the protein sample will gradually pass through areas of increasingly lower denaturant concentrations and higher pH, which promotes refolding into the native conformation. The shape and slope of the gradients, as well as the flow rate, will influence the refolding rate and can be adjusted for different protein samples. The principle is illustrated using a denatured recombinant scFv fusion protein obtained from E. coli inclusion bodies.

Centrifugation↗
PubMed1996

Nla and Nlb of peanut stripe potyvirus are present in the nucleus of infected cells, but do not form inclusions.

We investigated, by immunological and gene-fusion methods, whether the failure of peanut stripe potyvirus (PStV)-encoded nuclear inclusion proteins a (Nla) and b (Nlb) to form nuclear inclusions is due to the lack of their in vivo accumulation or the inability of one or both proteins to be transported into the nucleus Nla domains (Nla-VPg and Nla-proteinase), full-length Nlb, and full-length cylindrical inclusion (CI) protein of PStV were cloned, expressed in Escherichia coli, and used for antisera production. Immunoblot analysis of accumulation of Nla, Nlb, and CI in time course experiments revealed that they accumulated to similar levels in PStV-infected Nicotiana benthamiana. In immunocytochemical studies with electron microscopy, antiserum against Nla-VPg, Nla-Pro, and Nlb specifically labeled Nla and Nlb proteins throughout the nuclei of PStV-infected cells, in the absence of nuclear inclusions. Translational fusions were made between Nla and Nlb to either the green fluorescence protein or the beta-glucuronidase in vectors for transient gene expression or stable expression in transgenic plants respectively. Fusion proteins containing Nla accumulated in the nucleus, whereas fusion proteins containing Nlb accumulated in a punctate pattern in the cytoplasm. These data indicate that at least Nla possesses a nuclear localization signal.

Animals↗
PubMed2000

The p20 gene product of Citrus tristeza virus accumulates in the amorphous inclusion bodies.

Citrus tristeza virus (CTV) has 10 3' open reading frames (ORFs) of unknown function except for the two coat proteins. The highest produced subgenomic RNAs are those of the major coat protein gene (p25) and the 3' most genes, p20 and p23. The proteins from three ORFs, p25, p27, and p20, were examined in the yeast two-hybrid assay for the interactions between themselves and to one another. The p20 protein exhibited a high affinity for itself, suggesting that it might aggregate in infected cells. The cytopathology of CTV infections includes characteristic paracrystalline and amorphous inclusions in the phloem elements of infected citrus. Polyclonal antiserum raised against the bacterial expressed p20 gene product detected a protein of approximately 22-23 kDa, which accumulated to relatively high levels in CTV-infected citrus, but not in healthy citrus. Immunogold localization using antibodies to p20 protein showed strong and specific labeling of the amorphous inclusion bodies present in CTV-infected cells. Mesophyll protoplasts of Nicotiana benthamiana transfected with a CTV mutant containing the green fluorescent protein (GFP) ORF fused in-frame to the 3' end of p20 protein ORF expressed high levels of GFP. The fusion protein was concentrated in one specific area in the cytoplasm and lacked an organized shape. Accumulation of high levels of p20 protein in infected tissue, specific localization of the p20-GFP fusion protein, immunolocalization of p20 protein into amorphous inclusions, and strong homologous p20 protein-p20 protein interactions in the yeast-two-hybrid assay suggest that the p20 protein of CTV is a major component of the amorphous inclusion bodies present in CTV-infected cells.

Blotting, Western↗
PubMed1992

Biological variants of tobacco etch virus that induce morphologically distinct nuclear inclusions.

The presence of distinctive nuclear inclusions has been used for many years as a diagnostic character for tobacco etch virus (TEV). Cytological examinations of isolates of TEV in both weeds and solanaceous crops from areas widely separated geographically have demonstrated the presence of a variety of nuclear inclusions that vary considerably in form. Such inclusions can, in many cases, be related to differences in symptom expression. It is suggested that distinctive nuclear inclusions may be used to select biological variants of TEV that may be useful in the study and manipulation of closely related "strains" of this potyvirus.

Genetic Variation↗
PubMed1975

Annulate lamellae and crystalline inclusions in granular endoplasmic reticulum of the islet organ and associated tissues of a cyclostome, Myxine glutinosa.

Cytoplasmic annylate lamellae were found in the islet organ of a cyclostome, the hagfish (Myxine glutinosa), predominantly in cells interpreted as young proliferating beta-cells, and also in endocrine cells and enterocytes of the bile duct and gut and in the endothelial cells of small blood vessels. A close association was observed annulate lamellae and granular endoplasmic reticulum. Both in cells with and in those without annulate lamellae, crystalline inclusions of proteinaceous nature were seen in granular endoplasmic reticulum. These inclusions were occasionally closely associated to annulate lamellae, and a direct continuity could be seen between granular endoplasmic reticulum and the outer nuclear membrane surrounding an inclusion partially situated in the perinuclear cisterna. Rod-shaped structures and rounded electron dense bodies were seen in the nuclei of some islet parenchymal cells. The presence of annulate lamellae in the islet organ and associated tissues of Myxine glutinosa is believed to be related to the very high phylogenetic age of this species. The close association observed between annulate lamellae, granular endoplasmic reticulum, crystalline inclusions, and sometimes also nuclear membranes, may be of functional significance.

Animals↗
PubMed1978

Intranuclear inclusions in the developing neurons of the rat cuneate nuclei.

The ultrastructure of intranuclear rodlets, microtubules, fibrillar lattices and membranous inclusions found in the developing cuneate nuclei of rats is described. Rodlets, ranging in diameter from 96--312 nm and in length from 1--2 micrometer, are made up of tightly packed straight filaments measuring 5--8 nm in diameter. Microtubules with a diameter of 26 nm are clustered together. Fibrillar lattices are made up of fibrils with a diameter of 9 nm arranged in layers or sets. Two to nine sets make up a lattice, with a maximum width of 68 nm, in which the adjacent sets are arranged at an angle to each other. Rodlets and fibrillar lattices occur in 6.8% of the neurons. Membranous inclusions, reported here for the first time in normal neurons, are of 2 types: small vesicles of 0.1--0.6 micrometer and large vacuoles measuring 1--2 micrometer. Both types are bounded by either a single or a double membrane and generally have an electron lucent content. Membranous inclusions occur in 25.3% of the neurons. Changes in the frequency of occurrence of the various intranuclear inclusions in the course of postnatal development are also reported.

Animals↗
PubMed1994

A case of Pick's disease with unusual neuronal inclusions.

An autopsy case of unusual Pick's disease in a 61-year-old male is described. Findings included severe atrophy of the frontal and temporal lobes, pyramidal tracts and basal ganglia accompanied by numerous intraneuronal argyrophilic hyaline inclusions. His neurological symptoms were constantly progressive during the 12-year course, characterized by akinesia and emotional incontinence. The inclusions were round, well-demarcated, slightly eosinophilic and intensely argyrophilic bodies in the perikarya, and distributed mainly in the subiculum and Sommer's sector of the hippocampus, amygdala and affected gyri. Immunocytochemically, they contain antigenic determinants of both phosphorylated and nonphosphorylated neurofilaments, but were negative for ubiquitin. Ultrastructurally, they were composed primarily of skeins of neurofilaments intermingled with cell organelles. Tubular profiles studded with granular substances, previously reported as a feature of the generalized variant of Pick's disease, and Hirano body-like lattice structures were occasionally observed in the inclusions. This case represents a slowly progressive neurodegenerative disorder characterized by fronto-temporal lobar atrophy and might by categorized as a variant of Pick's disease. However, some unusual properties of neuronal inclusions may suggest a different pathogenesis from that in classical Pick's disease.

Dementia↗
PubMed1975

Cytoplasmic inclusions of neurons in the monkey visual cortex (area 19).

Several unusual neuronal inclusions were found in certain cells of the rhesus monkey visual cortex (Area 19): 1. Filamentous bodies, present in the small stellate cells of layer IV, globoid, 0.3-0.6 mum in diameter, consisting of fine 50 A filaments in a hexagonal meshwork. These are often associated with the labyrinthine bodies. 2. Labyrinthine bodies found exclusively in the small stellate cells of layer IV, including certain neurons with dispersed ribosomes. These are 0.4-0.7 mum in diameter and consist of 900 A wide tubes which interconnect with one another. The walls of these tubes are continuous and made up of a sheet or honeycomb lacework of small hexagonal 150 A subunits. 3. This inclusion, an aggregate 0.3-0.7 mum in size, consists of small membrane-bounded vesicles with a single dense granule associated with other non-membrane bound small dense droplets. The inclusions are always associated with the maturing face of the Golgi complex of certain layer IV pyramidal cells; as such, they may be an unusual product of the Golgi apparatus. These observations were confirmed by examination of stereo pairs of electron micrographs. Speculations are made with regard to possible functions for these inclusions.

Animals↗
PubMed1989

Intracytoplasmic eosinophilic inclusions in the neurons of the central nervous system.

This study reports the histological, ultrastructural, and immunocytochemical characteristics of intracytoplasmic eosinophilic inclusion bodies occurring in various types of neurons of the human central nervous system. By light microscopy, the inclusions were brightly eosinophilic, slightly birefringent, and sharply demarcated; they were found in the thalamus in 92% of the cases, in the substantia nigra in 88%, in the locus coeruleus in 45%, and rarely in the spinal cord. Ultrastructurally, the inclusions were composed of assemblies of parallel, alternating dark and light rectilinear profiles. The dark profiles corresponded to thin filaments (microfilaments) that measured 5.5-6.0 nm in diameter. A second set of dense lines crisscrossed the first at right angles. In sections perpendicular to their long axis, the filaments were distributed in a tetragonal lattice pattern in which individual elements occupied the angles of a square. Immunocytochemical preparations for actin were negative. Due to their high rate of occurrence in nonpathological brains, it is thought that the inclusions represent a normal but as yet unidentified cytoplasmic organelle.

Adult↗
PubMed1993

Strong immunoreactivity of alpha 1-antichymotrypsin co-localizes with beta-amyloid protein and ubiquitin in vacuolated muscle fibers of inclusion-body myositis.

In 10 of 10 inclusion-body myositis (IBM) patients, including 1 hereditary case, vacuolated muscle fibers contained large or small cytoplasmic inclusions immunoreactive for alpha 1-antichymotrypsin (alpha 1-ACT). All IBM muscle biopsies had characteristic cytoplasmic tubulo-filaments by electron microscopy. None of 17 control muscle biopsies contained the alpha 1-ACT immunoreactive inclusions characteristic of IBM. In vacuolated muscle fibers, alpha 1-ACT immunoreactive inclusions colocalized with beta-amyloid protein and ubiquitin immunoreactivities. Our study provides the first demonstration of alpha 1-ACT accumulations in abnormal human muscle, and it suggest that, as in Alzheimer's disease and Down's syndrome, alpha 1-ACT may be involved in the pathogenesis of IBM.

Adolescent↗
PubMed1976

[The neuronal Torres bodies in experimental yellow fever encephalomyelitis. An additional example of the heterogeneous nature of Cowdry type A intranuclear inclusions].

In experimental yellow fever virus encephalomyelitis of adult albino mice, the occurrence of intranuclear inclusions within nerve cells, in particular, within spinal and bulbar motoneurons, is a constant finding. The light microscopic features of these so-called neuronal Torres bodies correspond entirely to the criteria laid down by Cowdry for the type A intranuclear inclusions. At the ultrastructural level, these karyoplasmic formations consist of haphazardly distributed patches of an essentially amorphous material of various density. Unlike the Cowdry type A intranuclear inclusions characteristic of several herpesvirus and paramyxovirus infections, neuronal Torr-s bodies do not contain any viral structures. The frequency with which neuronal Torres bodies, certain nucleolar alterations and severe tigrolytic changes occur together, possibly indicates that this sort of type A intranuclear inclusion developes as a consequence of a virus induced disturbance of the cell nucleic acid and/or protein anabolism.

Animals↗
PubMed1986

Neuronal intranuclear hyaline inclusion disease in a nine year old.

Thirteen previous cases have been reported as neuronal intranuclear hyaline inclusion disease. The majority of patients have presented with movement disorders at less than 12 years of age followed by a progressive worsening of symptoms and, frequently, loss of cognitive function. Death has usually occurred by the second or third decade. Three have presented in the fifth through seventh decade with either movement disorders or dementia. These cases have been linked by the presence of eosinophilic neuronal intranuclear inclusions diffusely within the CNS and in peripheral ganglion cells. The patient in this case report also presented with a rapidly progressive movement disorder and at autopsy showed the characteristic intranuclear inclusions. Investigation of these inclusions did not reveal shared epitopes with neurofilaments or other intermediate filaments.

Brain↗
PubMed1989

Lewy body-like hyaline inclusions in sporadic motor neuron disease are ubiquitinated.

Round eosinophilic hyaline inclusion bodies with halos in the somata of anterior horn cells from a case of sporadic lower motor neuron disease (MND) were intensely immunostained with the monoclonal anti-ubiquitin antibody (DF2). A few similar, DF2-positive inclusions were also observed in the nerve cell processes of anterior horn cell or in the neuropil. Most inclusions showed intense homogeneous staining of the entire inclusion, whereas a few had intense staining of their periphery with no or pale staining of the central areas. Other DF2-positive structures in the somata of anterior horn cells included cytoplasmic granular structures, eosinophilic thread-like or reticular structures, and small eosinophilic profiles different from Bunina bodies. The DF2-staining intensity of Bunina bodies and spheroids did not exceed the background level. These results suggest that ubiquitination is associated with a pathological process of anterior horn cell degeneration in this MND case.

Aged↗
PubMed1984

Morphological characteristics of a unique intracytoplasmic neuronal inclusion body.

Unique intracytoplasmic inclusion bodies unaccompanied by a clinically apparent neurologic disease were found in the neurons of the hippocampus, subiculum, and mammillary body of a man who died suddenly, presumably of cardiac disease. The eosinophilic inclusions were round and often targetoid , with a central core that had the staining characteristics of RNA. Ultrastructurally, the inclusions were mainly composed of osmiophilic granular material. Rabies viral antigen was not detected by immunofluorescence, and heavy metals were not detected by energy dispersive spectrometry. These neuronal inclusions should be distinguished from Negri bodies.

Brain Diseases↗
PubMed1988

A new type of cytoplasmic inclusion in human choroid plexus. A histochemical, ultrastructural and frequency study.

We described a new type of cytoplasmic inclusion in the choroidal epithelial cells of humans. The inclusions usually appeared as brown, round or elongated bodies with or without an inner core, ranging in size from 1.3 to 7.0 micron. Histochemically, they contained polysaccharides, proteins and compound lipids. Ultrastructurally, they were composed of finely granular and filamentous materials, which are densely packed in the inner core and less dense in the outer zone. The frequency study of the inclusions in 197 autopsied patients revealed that their occurrence had no correlation with the age or the disease category. It is concluded that the inclusions are a nonspecific, but peculiar, change of the choroidal epithelial cells, probably representing the morphological expression of a physiological or pathological alteration of the cellular metabolism at the single-cell level.

Choroid Plexus↗
PubMed1979

Neuronal degeneration in the brain of the brindled mouse. II. Ultrastructure of neuronal inclusions in the cerebral cortex.

Two types of intra-cytoplasmic inclusions were observed in many cortical neurons of hemizygous brindled mutant mice (MObr/Y) which survived for more than 1 month. One type is an electron-lucent vacuole containing numerous vesicles and membrane fragments, some arranged concentrically, and the other consisted of electron-dense matrix with myelin figures. These inclusions were, as a rule, found near the axonal hillock where abnormally enlarged mitochondria were often present. In view of the presence of many features suggestive of autophagic vacuoles and absence of such inclusions in the oldest mice (91 days old), we hypothesize that these inclusions probably represent a focal repairing process of neurons from the sublethal injury.

Aneuploidy↗