scholarly journals Immunohistochemical Detection of Prion Protein in Sheep with Scrapie

1993 ◽  
Vol 5 (3) ◽  
pp. 309-316 ◽  
Author(s):  
Janice M. Miller ◽  
Allen L. Jenny ◽  
William D. Taylor ◽  
Richard F. Marsh ◽  
Richard Rubenstein ◽  
...  
Keyword(s):  
Prion Protein ◽  
Neuronal Cell ◽  
Normal Brain ◽  
Tissue Samples ◽  
Tissue Sections ◽  
Clinically Normal ◽  
Neuronal Cell Bodies ◽  
Histopathologic Evaluation ◽  
Transmissible Mink Encephalopathy ◽  
The Brain

Prion protein (PrP), which is involved in the pathogenesis of scrapie, occurs in 2 forms. The form extracted from scrapie brain is protease resistant (PrP-res), whereas PrP from normal brain is protease sensitive (PrP-sen). This study examined whether PrP-res could be detected in brains of sheep with scrapie by immunohistochemistry (IHC). A suitable IHC procedure was developed using brain tissue from hamsters that had been inoculated with the transmissible mink encephalopathy agent. Tissue samples were fixed in PLP (periodate, lysine, paraformaldehyde) that contained paraformaldehyde at a concentration of 0.125%. Before application of the IHC technique, tissue sections were deparaffinized and treated with formic acid to simultaneously enhance PrP-res immunoreactivity and degrade PrP-sen. Primary antibody was obtained from a rabbit immunized to PrP-res extracted from brains of mice with experimentally induced scrapie. Brains from 21 sheep with histopathologically confirmed scrapie were examined by IHC. In all 21 brains, PrP-res was widely distributed throughout the brain stem. Staining was particularly intense in neuronal cell bodies and around blood vessels. The IHC technique successfully detected PrP-res in brain samples that had been frozen or that were severely autolyzed before fixation in PLP. Brains from 11 scrapie-suspect sheep that were not considered histologically positive were also examined by IHC. PrP-res was found in 4 of these brains. Sections of brains from 14 clinically normal sheep did not have detectable PrP-res. Results of this study indicate that IHC detection of PrP-res is equivalent, and perhaps superior, to histopathology for the diagnosis of scrapie in sheep. Furthermore, IHC is applicable to tissues that have autolytic changes or processing artifacts that prevent satisfactory histopathologic evaluation for lesions of scrapie.

Metallic prions

2004 ◽  
Vol 71 ◽  
pp. 193-202 ◽  
Author(s):  
David R Brown
Keyword(s):  
Prion Protein ◽  
Binding Capacity ◽  
Normal Function ◽  
Transmissible Spongiform Encephalopathies ◽  
Published Data ◽  
Normal Brain ◽  
Copper Binding ◽  
Loss Of Function ◽  
Spongiform Encephalopathies ◽  
The Brain

Prion diseases, also referred to as transmissible spongiform encephalopathies, are characterized by the deposition of an abnormal isoform of the prion protein in the brain. However, this aggregated, fibrillar, amyloid protein, termed PrPSc, is an altered conformer of a normal brain glycoprotein, PrPc. Understanding the nature of the normal cellular isoform of the prion protein is considered essential to understanding the conversion process that generates PrPSc. To this end much work has focused on elucidation of the normal function and activity of PrPc. Substantial evidence supports the notion that PrPc is a copper-binding protein. In conversion to the abnormal isoform, this Cu-binding activity is lost. Instead, there are some suggestions that the protein might bind other metals such as Mn or Zn. PrPc functions currently under investigation include the possibility that the protein is involved in signal transduction, cell adhesion, Cu transport and resistance to oxidative stress. Of these possibilities, only a role in Cu transport and its action as an antioxidant take into consideration PrPc's Cu-binding capacity. There are also more published data supporting these two functions. There is strong evidence that during the course of prion disease, there is a loss of function of the prion protein. This manifests as a change in metal balance in the brain and other organs and substantial oxidative damage throughout the brain. Thus prions and metals have become tightly linked in the quest to understand the nature of transmissible spongiform encephalopathies.

scholarly journals Immunoreactivity of the calbindin D28k in the parahippocampal gyrus of chinchilla

2013 ◽  
Vol 57 (3) ◽  
pp. 387-391
Author(s):  
Radosław Szalak ◽  
Jadwiga Jaworska-Adamu ◽  
Karol Rycerz ◽  
Paweł Kulik ◽  
Marcin Bartłomiej Arciszewski
Keyword(s):  
Brain Area ◽  
Neuronal Cell ◽  
Immunofluorescence Staining ◽  
Parahippocampal Gyrus ◽  
Neuronal Cell Bodies ◽  
Entorhinal Area ◽  
Calbindin D28k ◽  
Retzius Cells ◽  
Species Specific ◽  
The Brain

Abstract Ten adult male chinchillas were used. The localisation of calbindin D28k (CB) was examined with the use of two types of reactions: immunocytochemical peroxidase-antiperoxidase and immunofluorescence staining with a specific monoclonal antibody against CB. Immunocytochemical examination demonstrated the presence of CB-positive neurons in the following layers of all parts the parahippocampal gyrus (PG): marginal, external cellular, middle cellular, and internal cellular, i.e. in entorhinal area, parasubiculum, and presubiculum. Immunofluorescence staining revealed the presence of CB in both Hu C/Dimmunoreactive (IR) neurons and nervous fibers of the PG. CB-IR neuronal cell bodies were moderately numerous (ca. 10% of Hu C/D-IR neurons) and clearly distinguished from the background. Each layer of the brain area consisted of two types of neurons: pyramidal and multiform. Among the second type of neurons, four kinds of morphologically different neuronal subclasses were observed: multipolar, bipolar, round, and Cajal-Retzius cells. It is concluded that the expression of CB in the PG of the chinchilla is species specific and limited to several subclasses of neurons

Early innervation of the metanephric kidney

Development ◽  
1988 ◽  
Vol 104 (4) ◽  
pp. 589-599 ◽  
Author(s):  
H. Sariola ◽  
K. Holm ◽  
S. Henke-Fahle
Keyword(s):  
Primary Cultures ◽  
Neuronal Cell ◽  
Similar Distribution ◽  
Renal Tubules ◽  
Experimental Conditions ◽  
Tissue Sections ◽  
Neuronal Cell Bodies ◽  
Metanephric Kidney ◽  
Kidney Morphogenesis

During kidney differentiation, the nephrogenic mesenchyme converts into renal tubules and the ureter bud branches to form the collecting system. Here we show that in the early undifferentiated kidney rudiment there is a third cell type present. In whole-mount preparations of cultured undifferentiated metanephric kidneys, neurones can be detected by immunohistochemical means with antibodies against the neurofilament triplet, 13AA8, and against neuronal cell surface gangliosides, Q211. Clusters of neuronal cell bodies can be seen in the mesenchyme close to the ureter bud. The terminal endings of neurites are found around the mesenchymal condensates that later become kidney tubules. A similar distribution of neurites can be revealed in tissue sections of kidney grafts growing in the chicken chorioallantoic membranes. In primary cultures of the ureter bud cells, neurones are constantly present. In another report, we have shown that, in experimental conditions, neurones are involved in regulation of kidney morphogenesis. The present results raise the possibility that neurones of the metanephric kidney may have this function in vivo as well.

Low-affinity sulfonylurea binding sites reside on neuronal cell bodies in the brain

1997 ◽  
Vol 745 (1-2) ◽  
pp. 1-9 ◽  
Author(s):  
Ambrose A Dunn-Meynell ◽  
Vanessa H. Routh ◽  
Joseph J McArdle ◽  
Barry E Levin
Keyword(s):  
Binding Sites ◽  
Neuronal Cell ◽  
Neuronal Cell Bodies ◽  
The Brain

scholarly journals SURFACE STRUCTURE OF ISOLATED NEURONS

1970 ◽  
Vol 47 (2) ◽  
pp. 319-331 ◽  
Author(s):  
Anders Hamberger ◽  
Hans-Arne Hansson ◽  
Johan Sjöstrand
Keyword(s):  
Electron Microscopy ◽  
Hypoglossal Nerve ◽  
Light And Electron Microscopy ◽  
Neuronal Cell ◽  
Spherical Particles ◽  
Hypoglossal Nucleus ◽  
Isolated Neurons ◽  
Tissue Sections ◽  
Neuronal Cell Bodies ◽  
Transmission Electron

Freehand, isolated neuronal perikarya from the hypoglossal nucleus of the rabbit have been examined with light-and electron-microscopy (transmission and scanning). The surface of the cell bodies was largely covered with spherical particles which were 0.5–2 µ in diameter. Transmission electron microscopy proved that the spherical particles were synaptic nerve terminals. Crush of the hypoglossal nerve which leads to chromatolysis and swelling of the neuronal cell bodies results in a conspicuous reduction in the number of terminals attached to the surface of hypoglossal neurons. This effect was observed both for isolated neurons and in tissue sections. The effect is considered in relation to earlier reported variations in the adherence of neuropil to isolated neuronal perikarya. The functional importance of nerve ending detachment in connection with nerve injury is discussed.

scholarly journals The prion protein gene: a role in mouse embryogenesis?

Development ◽  
1992 ◽  
Vol 115 (1) ◽  
pp. 117-122 ◽  
Author(s):  
J. Manson ◽  
J.D. West ◽  
V. Thomson ◽  
P. McBride ◽  
M.H. Kaufman ◽  
...  
Keyword(s):  
Gene Expression ◽  
Nervous System ◽  
Prion Protein ◽  
In Situ Hybridisation ◽  
Neuronal Cell ◽  
Normal Function ◽  
Cell Populations ◽  
Specific Expression ◽  
The Brain ◽  
Prp Gene

The neural membrane glycoprotein PrP (prion protein) has a key role in the development of scrapie and related neurodegenerative diseases. During pathogenesis, PrP accumulates in and around cells of the brain from which it can be isolated in a disease-specific, protease-resistant form. Although the involvement of PrP in the pathology of these diseases has long been known, the normal function of PrP remains unknown. Previous studies have shown that the PrP gene is expressed tissue specifically in adult animals, the highest levels in the brain, with intermediate levels in heart and lung and low levels in spleen. Prenatally, PrP mRNA has been detected in the brain of rat and hamster just prior to birth. In this study we have examined the expression of the PrP gene during mouse embryonic development by in situ hybridisation and observed dramatic regional and temporal gene expression in the embryo. Transcripts were detected in developing brain and spinal cord by 13.5 days. In addition, PrP gene expression was detected in the peripheral nervous system, in ganglia and nerve trunks of the sympathetic nervous system and neural cell populations of sensory organs. Expression of the PrP gene was not limited to neuronal cells, but was also detected in specific non-neuronal cell populations of the 13.5 and 16.5 day embryos and in extra-embryonic tissues from 6.5 days. This cell-specific expression suggests a pleiotropic role for PrP during development.

scholarly journals Phagocytic Activities of Reactive Microglia and Astrocytes Associated with Prion Diseases Are Dysregulated in Opposite Directions

Cells ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1728
Author(s):  
Anshuman Sinha ◽  
Rajesh Kushwaha ◽  
Kara Molesworth ◽  
Olga Mychko ◽  
Natallia Makarava ◽  
...  
Keyword(s):  
Prion Diseases ◽  
Neuronal Cell ◽  
Cell Types ◽  
Reactive Microglia ◽  
Neuronal Cell Bodies ◽  
Myelin Debris ◽  
Neurotoxic Effects ◽  
The One ◽  
The Brain ◽  
Phagocytic Uptake

Phagocytosis is one of the most important physiological functions of the glia directed at maintaining a healthy, homeostatic environment in the brain. Under a homeostatic environment, the phagocytic activities of astrocytes and microglia are tightly coordinated in time and space. In neurodegenerative diseases, both microglia and astrocytes contribute to neuroinflammation and disease pathogenesis, however, whether their phagocytic activities are up- or downregulated in reactive states is not known. To address this question, this current study isolated microglia and astrocytes from C57BL/6J mice infected with prions and tested their phagocytic activities in live-cell imaging assays that used synaptosomes and myelin debris as substrates. The phagocytic uptake by the reactive microglia was found to be significantly upregulated, whereas that of the reactive astrocytes was strongly downregulated. The up- and downregulation of phagocytosis by the two cell types were observed irrespective of whether disease-associated synaptosomes, normal synaptosomes, or myelin debris were used in the assays, indicating that dysregulations are dictated by cell reactive states, not substrates. Analysis of gene expression confirmed dysregulation of phagocytic functions in both cell types. Immunostaining of animal brains infected with prions revealed that at the terminal stage of disease, neuronal cell bodies were subject to engulfment by reactive microglia. This study suggests that imbalance in the phagocytic activities of the reactive microglia and astrocytes, which are dysregulated in opposite directions, is likely to lead to excessive microglia-mediated neuronal death on the one hand, and the inability of astrocytes to clear cell debris on the other hand, contributing to the neurotoxic effects of glia as a whole.

Ovine PrP transgenic Drosophila show reduced locomotor activity and decreased survival

2012 ◽  
Vol 444 (3) ◽  
pp. 487-495 ◽  
Author(s):  
Alana M. Thackray ◽  
Farooq Muhammad ◽  
Chang Zhang ◽  
Ying Di ◽  
Thomas R. Jahn ◽  
...  
Keyword(s):  
Locomotor Activity ◽  
Prion Protein ◽  
Germ Line ◽  
Critical Role ◽  
Neuronal Cell ◽  
Single Amino Acid ◽  
Site Specific ◽  
Protein Levels ◽  
Neuronal Cell Bodies ◽  
Transgenic Drosophila

Drosophila have emerged as a model system to study mammalian neurodegenerative diseases. In the present study we have generated Drosophila transgenic for ovine PrP (prion protein) to begin to establish an invertebrate model of ovine prion disease. We generated Drosophila transgenic for polymorphic variants of ovine PrP by PhiC31 site-specific germ-line transformation under expression control by the bi-partite GAL4/UAS (upstream activating sequence) system. Site-specific transgene insertion in the fly genome allowed us to test the hypothesis that single amino acid codon changes in ovine PrP modulate prion protein levels and the phenotype of the fly when expressed in the Drosophila nervous system. The Arg154 ovine PrP variants showed higher levels of PrP expression in neuronal cell bodies and insoluble PrP conformer than did His154 variants. High levels of ovine PrP expression in Drosophila were associated with phenotypic effects, including reduced locomotor activity and decreased survival. Significantly, the present study highlights a critical role for helix-1 in the formation of distinct conformers of ovine PrP, since expression of His154 variants were associated with decreased survival in the absence of high levels of PrP accumulation. Collectively, the present study shows that variants of the ovine PrP are associated with different spontaneous detrimental effects in ovine PrP transgenic Drosophila.

scholarly journals Detection of prion protein, the infection agent of bovine spongiform encephalopathy, by using aptamers

2019 ◽  
Vol 2019 (6) ◽  
pp. 5-8
Author(s):  
Алексей Антипин ◽  
Aleksey Antipin ◽  
Григорий Надточей ◽  
Grigoriy Nadtochey
Keyword(s):  
Prion Protein ◽  
Prion Proteins ◽  
Specific Interaction ◽  
Infectious Agent ◽  
Spongiform Encephalopathy ◽  
Dna Aptamers ◽  
Tissue Samples ◽  
Yeast Prions ◽  
The Brain ◽  
Brain Tissues

In the paper results of the specific interaction of selected DNA aptamers with prion protein from CNS tissues from DSE infected cows are described. The test detects amyloid epitops. The purpose of investigation. To study the possibility of detection of prion protein (BSE infectious agent in tissues CNS infected cows), using DNA aptamers. The DNA aptamers had been selected to yeast prions. Materials and methods. CNS tissues samples from a DSE experimentally infected cow were used. The infection was confirmed by histological analyses. The prion protein of BSE was isolated by ultracentrifugation in sucrose gradient and analysed, using immunoblotting and electron microscopy. Results. DSE infection in an experimentally infected cow was confirmed. The brain tissue samples were prepared. Specific effective binding of selected DNA aptamers with prions from DSE cow brain tissues was revealed. Conclusion. The possibility of detection of prion proteins in brain tissues from a DSE cow, using DNA aptamers, specifically binded with amyloid epitops was shown.

scholarly journals Enteric neuron density correlates with clinical features of severe gut dysmotility

2019 ◽  
Vol 317 (6) ◽  
pp. G793-G801 ◽  
Author(s):  
Elisa Boschetti ◽  
Carolina Malagelada ◽  
Anna Accarino ◽  
Juan R. Malagelada ◽  
Rosanna F. Cogliandro ◽  
...  
Keyword(s):  
Quantitative Analysis ◽  
Clinical Manifestations ◽  
Neuron Specific Enolase ◽  
Neuronal Cell ◽  
Enteric Neurons ◽  
Paraffin Sections ◽  
Tissue Samples ◽  
Neuron Density ◽  
Neuronal Cell Bodies ◽  
Gi Symptoms

Gastrointestinal (GI) symptoms can originate from severe dysmotility due to enteric neuropathies. Current methods used to demonstrate enteric neuropathies are based mainly on classic qualitative histopathological/immunohistochemical evaluation. This study was designed to identify an objective morphometric method for paraffin-embedded tissue samples to quantify the interganglionic distance between neighboring myenteric ganglia immunoreactive for neuron-specific enolase, as well as the number of myenteric and submucosal neuronal cell bodies/ganglion in jejunal specimens of patients with severe GI dysmotility. Jejunal full-thickness biopsies were collected from 32 patients (22 females; 16–77 yr) with well-characterized severe dysmotility and 8 controls (4 females; 47–73 yr). A symptom questionnaire was filled before surgery. Mann-Whitney U test, Kruskal-Wallis coupled with Dunn’s posttest and nonparametric linear regression tests were used for analyzing morphometric data and clinical correlations, respectively. Compared with controls, patients with severe dysmotility exhibited a significant increase in myenteric interganglionic distance ( P = 0.0005) along with a decrease in the number of myenteric ( P < 0.00001) and submucosal ( P < 0.0004) neurons. A 50% reduction in the number of submucosal and myenteric neurons correlated with an increased interganglionic distance and severity of dysmotility. Our study proposes a relatively simple tool that can be applied for quantitative evaluation of paraffin sections from patients with severe dysmotility. The finding of an increased interganglionic distance may aid diagnosis and limit the direct quantitative analysis of neurons per ganglion in patients with an interganglionic distance within the control range. NEW & NOTEWORTHY Enteric neuropathies are challenging conditions characterized by a severe impairment of gut physiology, including motility. An accurate, unambiguous assessment of enteric neurons provided by quantitative analysis of routine paraffin sections may help to define neuropathy-related gut dysmotility. We showed that patients with severe gut dysmotility exhibited an increased interganglionic distance associated with a decreased number of myenteric and submucosal neurons, which correlated with symptoms and clinical manifestations of deranged intestinal motility.

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