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J J Bernstein

Publications and source records attributed to J J Bernstein.

At least 19 recordsLinked to original sources
PubMed1999

Vascularization of human glioma spheroids implanted into rat cortex is conferred by two distinct mechanisms.

Aim of this study was to develop and characterize an applicable in vivo model to investigate angiogenesis of human gliomas. An established glioblastoma spheroid model was used to investigate the neovascularization of a standardized avascular solid tumor mass. Spheroids of two human glioma cell lines were labeled with an in vivo fluorescent dye. Single spheroids were implanted into the cortex of athymic rats. After 1, 3, 7, 14, and 21 days, brain sections containing the spheroid were immunostained for endothelial cells or vascular endothelial growth factor (VEGF). The dye-stained glioma spheroid and the endothelial cells were visualized by confocal microscopy. Two distinct mechanisms of tumor vascularization could be observed. (1) "Classical" angiogenesis with new vessels sprouting from existing host vessels into the spheroid was seen. (2) Individual endothelial cells were found to migrate towards and into the center of the spheroid where they coalesced to form new vessels. This process occurred as early as 24 hr after spheroid implantation. Spheroid vascularization was accompanied by an increase of VEGF expression, which peaked 7 days after implantation and returned to normal patterns by 14-21 days. Besides the "classical" angiogenesis by angiogenic blood vessels, the recruitment of individual endothelial cells seems to be an additional mechanism in early glioma vascularization. Our model proves to be a reliable, reproducible system to study in vivo angiogenesis of human gliomas.

Animals↗
PubMed1999

Cell-extracellular matrix interaction in glioma invasion.

Astrocytic tumours of the central nervous system express cell adhesion receptors of the integrin superfamily, CD44 and adhesion receptors of the immunoglobulin superfamily. Glioma cells utilize these receptors to adhere to and migrate along components of the extracellular matrix (ECM), which is uniquely distributed and regulated within the brain and the spinal cord. For penetration into healthy brain tissue a number of proteases are expressed, which degrade proteins of the extracellular matrix. Thus, glioma cell invasion into the adjacent brain tissue is dependent on the interaction of glioma cells with the extracellular matrix and the subsequent destruction of matrix barriers. There is a critical balance between expression of various adhesion receptors and proteases. The tight regulation of critical levels of proteases and receptors expressed by glioma cells or other cells is necessary for the "physiological" behaviour of glioma cells. Shifts in the balance of protein expression determine glioma cell behaviour in their micro-environment and can initiate or influence the complex process of glioma cell invasion. The complex receptor-ECM interaction in glioma cell invasion is discussed focussing upon the role of integrin receptors and matrix-metalloproteinases. Influencing these molecules or their regulation may lead to novel therapeutic approaches in the treatment of malignant glioma.

Animals↗
PubMed1998

ECM-mediated glioma cell invasion.

Cell adhesion receptors of the integrin superfamily, CD44, and adhesion receptors of the immunoglobulin superfamily are expressed by high-grade astrocytic tumors of the central nervous system. These receptors are critical for the invasion of these tumors in the nervous system. Glioma cells utilize these receptors to adhere to and migrate along the components of the extracellular matrix, which is uniquely distributed and regulated within the brain and the spinal cord. For this reason, glioma cell invasion into the adjacent brain tissue is dependent on the interaction of glioma cells with the extracellular matrix. The receptor-ECM component interaction is discussed, focusing on the role of cell adhesion molecules of the integrin family and CD44 in glioma cell adhesion and invasion.

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PubMed1997

Human A beta-amyloid and amyloid precursor protein accumulates in rat brain cells after cultured human leptomeningeal fibroblast implants.

Cultured human leptomeningeal fibroblasts grafted into rat frontal cortex were localized to the implant pocket and to adjacent host leptomeninges. Immunohistochemical studies using a panel of human-specific and domain-specific APP antibodies revealed that all grafted cells expressed both APP and A beta in situ. Remarkably, these antibodies also labeled rat pial and ependymal cells as well as reactive astrocytes adjacent to vessels. In addition, apical projections and cell bodies of many cortical pyramidal neurons contained human-specific APP immunoreactive material. Groups of subcortical neurons, particularly those of the amygdala, hippocampal formation and suprachiasmatic nuclei, were similarly labeled. The presence of human APP in host brains was confirmed by immunoblotting. Birefringent Congo Red staining was observed in the cortical neuropil and in leptomeningeal vessels. These data indicate that grafted leptomeningeal fibroblasts hyperexpress APP and A beta which can diffuse into parenchyma and be taken up by specific rat cells.

Amyloid beta-Peptides↗
PubMed1997

Implantation of cultured human leptomeningeal cells into rat brain.

Since previous studies have shown that cells cultured from human leptomeninges can express neuronal and glial antigens under appropriate culture conditions [DeGiorgio L. A. et al. (1994) J. Neurol. Sci. 124, 141 148; Bernstein J. J. et al. (1996) Int. J. Derl Neurosci. 14(5), 681 687], we have studied the developmental characteristics of these cells further by grafting them into young adult rat brains. Cells were labeled in culture with Fast Blue and were identified unequivocally by hybridization with nick-translated human DNA. Intensely Fast Blue positive human leptomeningeal cells were concentrated in the implant pocket and adjacent rat leptomeninges al one and two weeks postimplant. Human and rat leptomeningeal cells were similar morphologically and were equally immunopositive for vimentin and fibronectin. Implanted human cells did not express the neuronal and glial proteins they had in vitro. Cells which hybridized with human DNA corresponded to the intensely Fast Blue positive cells. Small groups of human DNA hybridizing cells were also observed in the choroid plexus. Less intensely Fast Blue positive neurons and glia were found in the brain but these hybridized with rat DNA. A minority of human leptomeningeal cells implanted into rat brain are subsequently found in host leptomeninges where they demonstrate properties characteristic of leptomeningeal fibroblasts. Small numbers of implanted cells can survive for two weeks.

Adult↗
PubMed1996

Changes in dynamin and actin mRNA expression in the dorsal column-medial lemniscal system following dorsal column lesion.

Transection of the third cervical hindlimb dorsal column nerve fibers in the spinal cord leads to a partial deafferentation atrophy of the neurons of the ascending dorsal column-medial lemniscal neural network (DC-ML) up to the cortex. We now examine the alteration of the steady-state level mRNA coding for the synaptic vesicle protein, dynamin I and the cytoskeletal protein beta-actin as early indicators of direct and trans-synaptic changes in the relay nuclei of the DC-ML. Rats were sacrificed at 6, 24, 72, and 240 hr after C3 hindlimb dorsal column or sham lesion. By 24 hr, there are changes in the steady-state levels of mRNA coding for both dynamin I and beta-actin in regions of the brain containing the first (nucleus gracilis of medulla) and third synaptic relays (cortex). Beta-actin mRNA is increased at both 6 and 24 hr in the nucleus gracilis. The changes in dynamin I mRNA in the nucleus gracilis are early and biphasic, elevated at 6 hr but decreased compared to sham by 24 hr. In both regions, the initial fluctuations of dynamin I and beta-actin mRNA levels are transient. By 72 hr, the levels are no different from those of sham-lesioned animals. In the somatomotor cortex, there is an additional increase in beta-actin mRNA levels at 240 hr. The increased steady-state levels of dynamin and actin mRNA following a hindlimb dorsal column lesion suggest that increased synaptic vesicle recycling and actin cytoskeleton rearrangement are some of the early responses to deafferentation made by the neurons of the DC-ML synaptic relays.

Actins↗
PubMed1996

Human leptomeningeal-derived cells express GFAP and HLADR when grafted into rat spinal cord.

The following series of experiments explores the post-xenografting differentiation of a naturally occurring, non-neuronal cell cultured from the leptomeninges of an 84-year-old woman. In culture, flat process-bearing human cells from the leptomeninges were positive for GFAP and 200 kDa neurofilament protein (negative for 68, 160 kDa neurofilament protein). The C3 spinal cord was exposed in 30 adult athymic rats. The hindlimb dorsal columns were transected at C3 and the nerve fibers aspirated to form a pocket, into which 10(6) fast blue-labeled, human leptomeningeal-derived cells were placed. The C3 spinal cord was studied immunohistochemically over 60 days. Three days later the dorsal horn contained fast blue-GFAP-positive astrocyte-like cells that were negative for neurofilament protein. By 7 days, large, process-bearing, fast blue-GFAP-positive (neurofilament protein-negative), astrocyte-like cells joined the native astrocytes of the pia-glia membrane and were in the gray matter of the spinal cord. Some of these astrocyte-like cells were also positive for the human specific histocompatibility complex, HLADR. These data extend the age, species and tissue of origin for pluripotential cells for CNS transplantation.

Adult↗
PubMed1996

Migrating fetal astrocytes do not intravasate since they are excluded from blood vessels by vital basement membrane.

Astrocytes appear to be excluded from blood vessels. In order to test this observation, pial blood vessels were seeded upon cultures of purified E14 fetal astrocytes. One to seven days later co-cultures were immunohistochemically stained for laminin and fibronectin. Confocal microscopy revealed that laminin-positive cultured astrocytes migrated from the coverslip to and on to the surface of the blood vessels. The fetal astrocytes migrated along the basement membrane but did not enter the blood vessel. Viable (live) basement membrane is a barrier to astrocyte intravasation and is a pathway for astrocyte migration.

Animals↗
PubMed1996

Absence of neuronal and glial proteins in human and rat leptomeninges in situ.

Human leptomeningeal (arachnoid and pia mater) cells in culture have been demonstrated in replicated studies to express typical neuronal proteins such as neurofilament protein and neuron-specific enolase. In addition, they can express glial fibrillary acidic protein. The present study examines the possibility that neuronal and glial proteins might be present in rat and human leptomeningeal cells in situ. The neuronal proteins 160 kDa and 200 kDa neurofilaments, neuron-specific enolase and microtubule-associated protein 2 were, however, not immunolocalized in either the pia mater or arachnoid. Glial fibrillary acidic protein and galactocerebroside were also not detected, while fibronectin and vimentin immunoreactivities were robust in all layers of the leptomeninges. Together with the previously reported expression of some neuronal and astroglial markers in cultured human leptomeninges, these observations suggest that culture alters the properties of leptomeningeal cells.

Animals↗
PubMed1995

Association of nerve growth factor mRNA levels with MK-801-induced explosive behaviors in mice.

MK-801, a noncompetitive antagonist of the N-methyl-D-aspartate (NMDA) receptor, stimulated an outbred strain of NIH Swiss mice to display discrete episodes of explosive jumping behavior, designated as "popping." The rapid onset of the MK-801-induced "popping" seems to follow the rapid distribution of the drug to the frontal cortex, the area that contains high levels of NMDA receptors. We examined the effect of this drug on the levels of mRNA coding for nerve growth factor (NGF) in the frontal cortex in relation to the exhibited "popping" episodes. Mice treated with 1 mg/kg MK-801 could be split into two groups based on the total number of "popping" episodes in a 30 min post-injection period. These groups also differed in the steady-state levels of frontal cortex NGF mRNA. Animals that exhibited low numbers of "popping" had levels of NGF mRNA significantly higher than saline treated controls or mice that exhibited high numbers of "popping." Mice treated with 10 mg/kg MK-801 had a high frequency of "popping" that was impossible to separate into episodes. In addition, these mice had levels of frontal cortex NGF mRNA that were significantly lower than either group of mice treated with 1 mg/kg MK-801. These data indicated that there was an increased level of NGF mRNA under conditions where MK-801 induced a low level of "popping" behavior. However, when "popping" intensified, NGF mRNA levels were decreased, suggesting a possible behavioral antagonism of the NGF response.

Animals↗
PubMed1995

Changes in c-fos expression in the dorsal column-medial lemniscal system following dorsal column lesions.

Transection of the hindlimb dorsal column fibers leads to a partial deafferentation of the neurons of the nucleus gracilis, the first relay of the ascending dorsal column-medial lemniscal (DC-ML) neural network. In response to the deafferentation, a synaptic renewal cycle is initiated and the neurons of the nucleus gracilis atrophy. The present study examines the molecular changes that occur in synaptic relays of the ascending DC-ML following a hindlimb dorsal column transection. Rats were sacrificed 0.5, 1, 24, or 72 hours postlesion. Steady-state levels of mRNA coding for c-fos were significantly elevated only at 24 hours postlesion in caudal dorsal medulla, which contains the nucleus gracilis. The increased c-fos is neuronal in origin since there is an increased level of c-fos immunoreactivity in both the cluster neurons and the interneurons of the nucleus gracilis. In the third relay of the DC-ML system, the somatomotor cortex, levels of c-fos mRNA were significantly decreased 72 hours postlesion. These data indicate that lesions of the hindlimb dorsal column fibers have transneuronal effects on gene expression that extend to at least the third synaptic relay in the DC-ML system.

Animals↗
PubMed1995

Glioblastoma cells do not intravasate into blood vessels.

Glioblastoma is very rarely found outside the central nervous system. The ability of rat C6 glioblastoma cells to intravasate into central nervous system and pial blood vessels is tested using a rat homografting model and two in vitro models. In vivo, scanning electron microscopy demonstrates that upon grafting C6 cells into implantation pockets in rat cortex, blood vessels can be spared in large digestion cysts formed in host brain parenchyma. Immunocytochemistry of the grafted rat cortex reveals that the glioblastoma cells are upon the blood vessel basement membrane, surrounded by the extracellular matrix material, fibronectin. The endothelial cells of the blood vessel are inside the laminin and fibronectin, and there were areas of endothelial cell hyperplasia. C6 cells are not observed inside blood vessels. In vitro, C6 cell cultures seeded with blood vessels from fresh rat pia exhibit the same relationship of the C6 glioblastoma cells to the blood vessel as those in the other models. The C6 cells migrate upon the pial blood vessel basement membrane but do not intravasate into the blood vessel. To ascertain whether structure and components of the blood vessel basement membrane are important factors in glioblastoma cell exclusion from blood vessels, C6 cells are seeded upon artificial basement membrane hydrated gel wafers. C6 cells migrate into the artificial basement membrane gel wafer by 1 day after seeding. These data indicate that glioblastoma cells are confined to the central nervous system by an inability to pass through vital basement membrane.

Animals↗
PubMed1994

Migration of fresh human malignant astrocytoma cells into hydrated gel wafers in vitro.

Individual astrocytoma cells expressing a cytoplasmic form of p185c-neu migrated along basement membrane lined surfaces after xenografing fresh low or high grade human malignant astrocytomas into host rat brain. We now study the migratory capacity of fresh human malignant astrocytoma cells seeded on hydrated gel wafers composed of artificial basement membrane or collagen I, a normal and lesion-related CNS extracellular matrix component. Approximately 10(7) mechanically disrupted cells (with small clumps) of 3 fresh low grade and 6 fresh high grade astrocytomas were seeded on the surface of artificial basement membrane and collagen I wafers (11 x 16 mm). The wafers were then prepared for scanning electron microscopy and immunohistochemistry at 1, 3, 5, and 7 days after seeding. Regardless of tumor grade, a morphologically similar class of cells was observed to migrate through collagen I gels in 24 hours and 0.5-1.5 mm into artificial basement membrane gels in 7 days. Immunohistochemistry revealed that the migrated cells from low and high grade astrocytomas were positive for glial fibrillary acidic protein (GFAP)) and expressed cytoplasmic human-specific p185c-neu. These data indicate that fresh human malignant astrocytoma cells that contain GFAP and express cytoplasmic p185c-neu have a high degree of migratory capacity and could be the cell in the tumor involved in intraparenchymal metastasis and poor patient survival in high grade astrocytomas of the human brain.

Astrocytoma↗
PubMed1994

Canine distemper epizootic in lions, tigers, and leopards in North America.

Canine distemper virus (CDV) infection occurred in captive leopards (Panthera pardus), tigers (Panthera tigris), lions (Panthera leo), and a jaguar (Panthera onca) in 1991 and 1992. An epizootic affected all 4 types of cats at the Wildlife Waystation, San Fernando, California, with 17 mortalities. CDV-infected raccoons were thought to be the source of infection in these cats. Two black leopards died at the Naibi Zoo, Coal Valley, Illinois, and 2 tigers died at the Shambala Preserve, Acton, California. Initial clinical signs were anorexia with gastrointestinal and/or respiratory disease followed by seizures. Canine distemper virus was isolated from 3 leopards, 3 tigers, and 3 lions that died or were euthanized when moribund. Monoclonal antibody testing identified the virus isolates as CDV. Gross and histopathologic findings were similar to those found in canids with distemper with a few exceptions. There were fewer lesions in the brain, and there was a pronounced type 2 cell proliferation in the lung, with inclusion bodies and CDV antigen demonstrated by immunohistology. Neutralizing antibody to CDV was found in high titers in serum from most animals but was absent or was found only in low titers in some cats that succumbed after CDV infection. There was a marked difference in neutralizing antibody titers when tests were done with different strains of CDV.

Animals↗
PubMed1993

Migrated fetal astrocytes modulate nerve growth factor expression in host nucleus gracilis of the medulla after grafting in third cervical hindlimb dorsal columns of the spinal cord.

Nerve growth factor (NGF) immunoreactivity in the nucleus gracilis of the medulla was quantitated for 90 days after aspiration of the C3 spinal hindlimb dorsal columns of 36 adult rats. Half the lesioned animals were a lesion-only group. The remaining lesioned animals received an immediate graft of two 1.0-mm pieces of 14 day gestation fetal rat cervical spinal cord (prelabeled with Phaseolus vulgaris leucoagglutinin) into the aspiration pocket (graft group). There were 3 normal controls. Groups of animals were analyzed at 7, 14, 21, 30, 60, and 90 days. At 90 days, NGF immunoreactivity was significantly elevated in the nucleus gracilis of lesion-only animals. This increase in NGF immunoreactivity was augmented in glial end-feet surrounding neurons and was also observed in the cytoplasm of astrocytes and some neurons. Previous experiments have shown that the cluster neurons of the nucleus gracilis undergo atrophy at this time with a concomitant decrease in hindlimb placement. NGF immunoreactivity (90 days) in grafted animals, however, was significantly less than in lesion-only animals (P < 0.05) but remained significantly elevated above control animals (P < 0.05). Unlike in lesion-only animals, there were no NGF positive neurons in the nucleus gracilis of grafted animals. Previous experiments have shown that astrocytes from fetal spinal cord grafts migrate to the nucleus gracilis, maintain cluster neuron cell size, and improve hindlimb placement at 90 days. The present data indicate that modulation of detrimental increases in NGF appeared to be a mechanism by which migrated fetal astrocytes can be used as a system for cell therapy.

Animals↗
PubMed1993

Migration of human malignant astrocytoma cells in the mammalian brain: Scherer revisited.

Fresh suspensions of human glioblastoma multiforme were preincubated in the plant lectin Phaseolus vulgaris leucoagglutinin (PHAL) and implanted into cortical pockets in adult rat brain. Brains were investigated periodically over 30 postoperative days and the migration of the human glioblastoma cells was traced with anti-PHAL immunofluorescence or the overexpression of human specific p185c-neu a specific marker of a class of human malignant astrocytoma cells. The principal pathway of migration of the implanted human cells in the rat brain was ventrally through cortical gray matter and into the corpus callosum, with rapid lateral distribution in this and other parallel and intersecting white matter fascicles. Human glioblastoma cells also migrated on basement membrane lined blood vessels, pia-glia membrane and spaces of Virchow-Robin, as well as the subependymal space of the ventricles. These paths of migration of human glioblastoma cells in the rat brain are consistent with the pathways of spread of glioblastoma in the human brain as described by Scherer over 50 years ago, indicating that multifocal malignant astrocytomas have common migratory pathways in mature mammalian brain.

Animals↗