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R I Carp

Publications and source records attributed to R I Carp.

At least 73 records · Page 4Linked to original sources

Incubation periods and histopathological changes in mice injected stereotaxically in different brain areas with the 87V scrapie strain.

After stereotaxic injection into five different brain areas (cortex, caudate nucleus, substantia nigra, thalamus and cerebellum) of IM mice with the 87V scrapie strain, the cerebellum had the shortest incubation period. The vacuolation pattern was similar regardless of the area injected with extensive vacuolation in the thalamus, mesencephalon and hypothalamus. The pattern of amyloid plaques differed markedly depending on the area injected. In particular, no plaques were seen anywhere in the brain after injection into intact cerebellum, whereas injection into the four cerebral areas yielded plaques in the forebrain but not in the cerebellum. The incubation period after injection into bisected cerebella was much longer than after injection into intact cerebella. Mice injected on one side of bisected cerebellum had amyloid plaques in the forebrain but not in the cerebellum. There is a discussion of the finding that, although no plaques and virtually no vacuolation were seen in the cerebellum, the shortest incubation period occurred after injection into intact cerebellum.

Amyloid↗

Pancreatic lesions and hypoglycemia-hyperinsulinemia in scrapie-injected hamsters.

Hamsters injected intracerebrally with scrapie strains 139H or 22CH or with normal hamster brain were assessed for body weight periodically throughout the incubation period. Animals injected with the scrapie strains became obese before the appearance of the motor changes that are indicative of the start of clinical disease. During the latter part of the incubation period and during clinical disease, animals were hypoglycemic and showed marked hyperinsulinemia. At autopsy, there was marked hyperplasia and hypertrophy of the cells of the islets of Langerhans. Thyroid, adrenal glands, liver, and kidney also were enlarged. The data suggest a severe endocrinopathy and point to new areas of pathologic and clinical changes that can be assigned to unconventional slow infections.

Animals↗

Vacuolization, incubation period and survival time analyses in three mouse genotypes injected stereotactically in three brain regions with the 22L scrapie strain.

In previous studies we showed that C57BL mice injected stereotactically in the cerebellum with the 22L scrapie strain had a significantly shorter incubation period than those injected with the same agent in other brain regions. In mice injected in the cerebellum, vacuolization was limited to the cerebellum, medulla and mesencephalon, whereas injection into forebrain regions resulted in vacuolization in all brain regions. The studies suggested that the cerebellum had a selective vulnerability for 22L. In this study we examined the interaction between host genotype and selective vulnerability of specific brain regions. The mouse gene that has the most profound effect on pathogenesis, particularly incubation period, is termed Sinc (scrapie incubation). Groups of mice with three genotypes of Sinc (s7s7, p7p7 and their F1 cross, s7p7) were injected with 22L into the cerebral cortex, thalamus or cerebellum. Analysis of incubation periods showed that, regardless of the host genotype, the cerebellum injection group had a significantly shorter incubation period than groups injected in other regions. After cerebellum injection vacuolization was limited to the cerebellum, medulla and mesencephalon in all three host genotypes. The location of vacuoles within the cerebellum differed depending upon the host strain. Vacuolization developed almost exclusively in grey matter in s7s7 mice, mainly in white matter in p7p7 mice, and in both grey and white matter in F1 mice. These results demonstrate that the selective vulnerability of the cerebellum to induction of clinical disease by 22L does not depend on host genotype, but host genotype does affect lesion distribution within the cerebellum.

Animals↗

Pathogenesis and pathology of scrapie after stereotactic injection of strain 22L in intact and bisected cerebella.

The mechanisms involved in the spread of scrapie within the brain remain unclear. To examine this issue the 22L scrapie strain was injected in one side of the cerebellum of mice in which the cerebellum had been bisected prior to injection. Another group of animals received the same injection into intact cerebella, i.e. without prior bisection. We found that bisection of the cerebella delayed the spread of scrapie agent from the injection site to the contralateral side of the cerebellum and that the occurrence of vacuolization was not as extensive and was markedly delayed in the uninjected side compared to its occurrence after injection in the intact cerebellum. Replication of agent in an area preceded the development of vacuolization in that area by several weeks. There was marked loss of Purkinje cells on the injected side of bisected cerebella, with no loss seen on the uninjected side. The incubation period of scrapie disease in mice injected after cerebellar bisection was significantly longer than after the injection of intact cerebella. The results in this study suggest that the scrapie agent spreads along intact nerve cell tracts, probably by axonal transport.

Animals↗

Further characterization of scrapie replication in PC12 cells.

The rat pheochromocytoma cell line, PC12, undergoes neuron-like morphological, biochemical and electrophysiological differentiation, in the presence of low concentrations of nerve growth factor (NGF). NGF-treated PC12 cells have been shown previously to support 139A scrapie agent replication. In the present report we extended these findings and analysed the cellular conditions necessary for agent replication. Following the infection of differentiated PC12 cells, scrapie replicated to relatively high titres as determined by an incubation period assay. The removal of NGF, which causes the gradual dedifferentiation of PC12 cells, resulted in the inability of scrapie to replicate. The scrapie infectivity detected in PC12 cultures is cell-associated and not released into the medium. Cells in infected cultures did not show any change in morphology when compared to cells in mock-infected cultures. Titration studies of scrapie infectivity in PC12 cells have indicated that up to 4 LD50 units per cell can be obtained although a yield of 1 LD50 per cell was more common. Using an approximate m.o.i. of 1, only differentiated PC12 cells supported 139A scrapie agent replication when compared to two other differentiated, neuronal cell types, indicating that PC12 cells are more susceptible to agent replication. These studies support further the suitability of using differentiated PC12 cells as an in vitro model to study scrapie agent replication.

Adrenal Gland Neoplasms↗

Scrapie-induced alterations in glucose tolerance in mice.

Certain scrapie strains cause obesity in several strains of mice. The potential association between obesity and altered glucose tolerance was assessed by monitoring body weight and glucose tolerance throughout the incubation period in scrapie strain-mouse strain combinations that do and do not produce obesity. Virtually all obese mice showed reduced glucose tolerance as shown by significantly higher blood glucose levels 2 h after a glucose overload. Mice injected with a scrapie strain that did not cause obesity showed normal tolerance. The scrapie infectivity titre of the pancreas of obese mice clinically affected with scrapie was very low. Adrenalectomy prevented both the increase in weight and aberrant glucose tolerance but had no other effect on the course of the disease. Following increasing dilution of the inoculum, the increase in body weight and the development of aberrant glucose tolerance reached an end-point that was similar to that of scrapie infectivity. The system described provides an inducible model of obesity with altered glucose tolerance.

Adrenalectomy↗

Classic genetics of scrapie.

Many years ago, observations of natural scrapie revealed differences in clinical manifestations and in the areas of the brain that showed the most intense histopathological changes. In experimental work with scrapie in mice two fundamental points were established in early studies: (1) A mouse gene, termed Sinc for scrapie incubation, affects the length of the incubation period. (2) In a single strain of mice, incubation periods of different "strains" of scrapie can differ by as much as 3-4 fold, e.g., the incubation period of the ME7 scrapie strain in C57BL mice is approximately 122 days, whereas the incubation period for the 22A strain is 360 days. From this "classic" beginning our knowledge about the parameters influenced by genetic characteristics of both host and agent have expanded. In a number of instances, initial examination of an aspect of host-agent interaction has subsequently revealed surprising influences of informational molecules from both host and agent. A partial list of the parameters under genetic control follows: (1) The extent of vacuolation in various regions of the brain; (2) The quantity of amyloid plaques in the brain; (3) The occurrence of increased weight in the preclinical phase of disease; (4) The development of aberrant glucose tolerance; (5) The area of the brain which yields the shortest incubation period after stereotaxic injection; and (6) The physical-chemical and immunological characteristics of scrapie associated fibrils. In several well documented instances differences between scrapie strains were evident after repeated passages of the strains in the same host strain.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Influence of stereotaxically injected scrapie on neurotransmitter systems of mouse cerebellum.

The 22L strain of scrapie was injected stereotaxically into the cerebellum of C57BL/6J mice to determine its effect on several cerebellar neurotransmitter systems during the early clinical stages of the disease. In this model vacuolar lesions are restricted to the cerebellum with no evidence of vacuolization in other brain regions. Although vacuolar lesions develop throughout all cell layers of the cerebellum, they are most severe in the granule cell layer. Modest but significant (P less than 0.01) reductions in cerebellar weight, glutamate decarboxylase activity, and in the affinity of the N6-[adenine-2,8-3H]cyclohexyladenosine binding sites, were observed in scrapie affected mice. The densities of the high- and low-affinity adenosine receptors were unaffected. Adenosine receptors in the cerebellum are highly localized to the axon terminals of the glutamatergic, GABA receptive granule cells. GABA, benzodiazepine, glutamate, and muscarinic cholinergic receptors were not significantly altered. In addition, the high-affinity uptake of glutamate, and the activity of choline acetyltransferase were not significantly changed. GABA high-affinity uptake was slightly increased. Even though the granule cell layer of the cerebellum had undergone severe vacuolation, only modest neurotransmitter changes were apparent. Although these results suggest a tenuous relationship between scrapie pathology and the integrity of neurotransmitter systems, it is possible that compensatory neurochemical changes in uncompromised neuronal populations may have masked potentially specific neurotransmitter effects.

Animals↗

Multiple sclerosis etiology--an Epstein-Barr virus hypothesis.

The opinion has been expressed frequently in the literature that MS is caused by a combination of an environmental agent and genetic factors, possibly with an autoimmune component. However, the specific environmental agent has not as yet been identified. This paper describes cumulative circumstantial evidence which strongly points to a post-pubertal primary Epstein-Barr virus infection as this unidentified etiological event in the induction of MS disease.

Adolescent↗

Adrenal involvement in scrapie-induced obesity.

In previous studies we found an increase in body weight during the preclinical phase of disease in certain scrapie strain-mouse strain combinations. The effect was augmented by injection into the hypothalamus. In the present study, we found an increase in food consumption (compared to the normal mouse brain injection group) for both the 139A and ME7 scrapie groups, although only the ME7 group showed an increase in body weight. In a scrapie strain-mouse strain combination that showed an increase in body weight, the adrenal gland was the only organ that showed a significant increase in weight. The titer of scrapie in the adrenals was comparatively low. Adrenalectomy prevented the increase in body weight in two strains of mice injected with the ME7 scrapie strain. The results suggest that scrapie-induced obesity depends on an effect of scrapie on the hypothalamic-pituitary-adrenal axis.

Adrenal Glands↗

Ultrastructural observations of spinal cord lesions and blood-brain barrier changes in scrapie-infected mice.

Spinal cord samples from IM or VM mice injected intracerebrally with the 87V scrapie agent were examined ultrastructurally at the clinical stage of disease for changes in blood vessel permeability and for pathological alterations. In several animals, (3 of 16), massive changes were noted in the cervical spinal cords in the subependymal area of the cortical gray matter immediately surrounding the central canal including ependymal cell changes, the presence of amyloid plaque in close association with microglial cells, extensive neuropil vacuolation, the appearance of reactive astrocytes, degenerating neurites and vacuolated neurons. In those regions showing structural damage, localized increased permeability to horseradish peroxidase across the blood-brain barrier was noticed along with the appearance of numerous vesiculo-canalicular profiles in micro-blood vessel endothelial cells with extravasation of the tracer to the neuropil. Some damaged neurons appeared flooded with this tracer. These changes were not observed in either the thoracic or lumbar spinal cord regions. The occurrence of pathological changes in the spinal cords of a small percentage of intracerebrally injected mice was probably due to a high concentration of the scrapie agent which localized in the cervical spinal cord, presumably after entering the spinal fluid via the lateral ventricle at the time of injection.

Amyloid↗

Detection of scrapie-associated fibrils (SAF) and SAF proteins from scrapie-affected sheep.

Scrapie-associated fibrils (SAF) were detected by negative-stain electron microscopy in the brains (by two different isolation procedures) and spleens of sheep naturally and experimentally infected with scrapie. Although the numbers of SAF varied from case to case, the yield of SAF from brains of naturally affected sheep was lower than that from experimentally affected sheep. SAF-specific, protease-resistant proteins (PrPs) were detected by silver staining and western blot analysis in most samples of brain from experimentally affected sheep. PrPs, however, could be detected in only a limited number of natural cases of sheep scrapie because of the lower yields of SAF. PrPs from sheep SAF appear biochemically and antigenically similar to PrPs from other species infected with unconventional agents. This study further establishes the unique association of SAF and PrPs with natural or experimentally induced scrapie in its natural host.

Animals↗

Genetic control of scrapie: incubation period and plaque formation in I mice.

The host component of control of scrapie incubation period in the mouse is manifested largely through the action of the Sinc gene. Only one mouse strain (VM) has been found that is p7p7 (prolonged incubation for ME7 agent) and two other strains have been derived from VM. All other strains, designated s7s7, have a short incubation for ME7. In the present study, the I strain was shown to fulfil the criteria that are characteristic of mouse strains with the p7 allele of Sinc: a comparatively long incubation period for ME7 and a short incubation period for 22A, the incubation period for F1 hybrid mice (s7s7 X p7p7) either fell between the incubation periods for the parental strains (with ME7) or were longer than either parent (with 139A and 22A), amyloid plaques occurred following injection of ME7 and 87V but not after 22A or 139A, lesion profiles for four scrapie strains were similar in I mice and p7p7 mouse strains, and injection of 87V led to disease in less than 300 days. Finally, allelism tests using F1 hybrid mice (I X a p7p7 mouse strain) and progeny of backcrosses between these F1 mice and I mice failed to reveal the segregation of additional major genes affecting scrapie incubation period.

Alleles↗

Incubation periods and survival times for mice injected stereotaxically with three scrapie strains in different brain regions.

Incubation period and survival time were determined in C57BL mice which had been injected stereotaxically with either the 139A, ME7 or 22L strain of scrapie in one of five different brain regions (cerebral cortex, caudate nucleus, thalamus, substantia nigra, cerebellum). The injection of 139A in the caudate nucleus, thalamus, substantia nigra or cerebellum resulted in significantly shorter incubation periods than following cerebral cortex injection. For ME7, mice injected in the thalamus and cerebellum had incubation periods approximately 2 weeks shorter than the cerebral cortex group. For 22L, the incubation periods after substantia nigra or cerebellum injection were significantly shorter than after cortex injection. The cerebellum injection group had a significantly shorter incubation period than the substantia nigra injection group. The survival times for mice injected with a particular scrapie strain were directly related to the incubation periods. The groups with the shortest incubation also had the shortest survival time (e.g. 22L in the cerebellum). On histological examination, 139A and ME7 produced brain lesions in all brain areas regardless of injection site. For the 22L strain, after cerebellum injection vacuolation was limited to the cerebellum, while injection into the cerebral cortex and other forebrain regions resulted in vacuolation in all brain regions examined. Despite the difference in the distribution of vacuolation seen in cerebellum- compared to cortex-injected (22L) mice, infectivity titres were similar in the cortex, cerebellum, cerebellar cortex and medulla plus pons.

Animals↗

Antisera to scrapie-associated fibril protein and prion protein decorate scrapie-associated fibrils.

Scrapie-associated fibrils (SAF) are an infection-specific structure observed in the unconventional-agent diseases. Polyclonal antisera raised to scrapie proteins were used to test the antigenic relationship between purified fibrils and SAF isolated from non-protease-treated synaptosomal-mitochondrial preparations. The experimental design utilized fibrils from scrapie strain 263K-infected hamsters, scrapie strain 139A-infected mice, and scrapie strain ME7-infected mice. Preparations were examined by negative-stain immune electron microscopy and Western blot analysis of the polypeptides. Fibrils and polypeptides from each preparation reacted with a rabbit antiserum raised to each of the following: hamster 263K prion protein (PrP 27-30), hamster 263K SAF protein, and mouse ME7 SAF protein. Immune electron microscopy and Western blot analysis revealed similar antigenic relationships among the three scrapie antisera. Thus, fibrils and polypeptides can be considered to be the same in each preparation. No reactivity of the fibrils was observed with antisera raised to Alzheimer neurofibrillary tangles or a synthetic peptide of cerebrovascular amyloid. Thus, the fibrils observed in purified preparations share structural and antigenic similarities plus biochemically related peptides with SAF present in non-protease-treated preparations.

Animals↗

Mouse polyclonal and monoclonal antibody to scrapie-associated fibril proteins.

Antibody response in mice to scrapie-associated fibril proteins (protease-resistant proteins [PrPs]) was generated to different epitopes depending on the source of antigen. Mice responded differently to PrPs isolated from scrapie-infected animals of homologous (mouse) versus heterologous (hamster) species. An enzyme-linked immunosorbent assay established to monitor this antibody response in mice immunized with PrPs was unable to detect such a response in scrapie-infected mice. A monoclonal antibody (MAb), 263K 3F4, derived from a mouse immunized with hamster 263K PrPs reacted with hamster but not mouse PrPs. MAb 263K 3F4 also recognized normal host protein of 33 to 35 kilodaltons in brain tissue from hamsters and humans but not from bovine, mouse, rat, sheep, or rabbit brains. This is the first demonstration of epitope differences on this host protein in different species. The defining of various epitopes on PrP through the use of MAbs will lead to a better understanding of the relationship of PrPs to their host precursor protein and to the infectious scrapie agent.

Animals↗

Paired helical filaments associated with Alzheimer disease are readily soluble structures.

Considerable controversy exists concerning the origin and composition of Alzheimer neurofibrillary tangles (ANT) and of paired helical filaments (PHF), the abnormal cytoplasmic fibers which ultrastructurally are the major components of ANT. Thus far, the unusual solubility properties of PHF have hindered the analysis of ANT. A new procedure is presented for isolating purified PHF which are soluble in the presence of sodium dodecyl sulfate. The purification protocol involves differential and rate zonal centrifugation, treatment with the detergents sarcosyl and sulfobetain 3-14, and sonication. The isolated PHF from Alzheimer disease/senile dementia of the Alzheimer type (7 cases) and Down's syndrome (one case) have been characterized structurally by negative-stain electron microscopy, biochemically by PAGE, and immunologically by both the ELISA technique and Western blot analysis using a monoclonal antibody prepared against ANT. Distinct polypeptides were shown to be associated with this structure and not seen in preparations from young and age-matched normal brains.

Adult↗