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Biomedical subjects

S W Barger

Publications and source records attributed to S W Barger.

At least 37 records · Page 2Linked to original sources
PubMed1996

Participation of gene expression in the protection against amyloid beta-peptide toxicity by the beta-amyloid precursor protein.

The amyloid beta-peptide (A beta) is a toxic derivative of the beta-amyloid precursor protein. Alternative processing of this precursor also yields large soluble forms (APPSs) which are secreted from many cell types. These APPSs have neuritogenic and neuroprotective activities; indeed, APPSs can protect primary neurons from the toxicity of A beta itself. To begin to explore the regulation of gene expression by APPS, we have focused on the NF-kappa B transcription factor family. NF-kappa B is induced by conditions of stress, including cellular oxidation. We report that NF-kappa B can also be induced by APPS. Furthermore, we effected direct activation of NF-kappa B through disinhibition using antisense oligonucleotide technology. This means of activating NF-kappa B resulted in protection of neuroblastoma cells from the toxicity of a calcium ionophore and protection of primary hippocampal neurons from the toxicity of A beta. Together, these data suggest that NF-kappa B may exist as a common agent inducing a neuroprotective pattern of gene expression in response to either trophic cytokines or stress itself.

Amyloid beta-Peptides↗
PubMed1996

Activation of K+ channels and suppression of neuronal activity by secreted beta-amyloid-precursor protein.

The Alzheimer's beta-amyloid precursor protein (beta-APP) is widely expressed in neural cells, and in neurons secreted forms of beta-APP (sAPPs) are released from membrane-spanning holo-beta APP in an activity-dependent manner. Secreted APPs can modulate neurite outgrowth, synaptogenesis, synaptic plasticity and cell survival; a signal transduction mechanism of sAPPs may involve modulation of intracellular calcium levels ([Ca2+]i). Here we use whole-cell perforated patch and single-channel patch-clamp analysis of hippocampal neurons to demonstrate that sAPPs suppress action potentials and hyperpolarize neurons by activating high-conductance, charybdotoxin-sensitive K+ channels. Activation of K+ channels by sAPPs was mimicked by a cyclic GMP analogue and sodium nitroprusside and blocked by an antagonist of cGMP-dependent kinase and a phosphatase inhibitor, suggesting that the effect is mediated by cGMP and protein dephosphorylation. Calcium imaging studies indicate that activation of K+ channels mediates the ability of sAPPs to decrease [Ca2+]i. Modulation of neuronal excitability may be a major mechanism by which beta-APP regulates developmental and synaptic plasticity in the nervous system.

Action Potentials↗
PubMed1996

Induction of neuroprotective kappa B-dependent transcription by secreted forms of the Alzheimer's beta-amyloid precursor.

A significant fraction of the beta-amyloid precursor protein is proteolytically processed to yield large secreted forms (sAPP). These proteins have pleiotropic effects which potentially involve control of gene expression. We have investigated the influence of sAPP on the class of transcription factors which bind kappa B enhancer sequences. Transcription dependent on a kappa B element was enhanced by sAPP in several cell lines, as measured by expression of a transfected chloramphenicol acetyltransferase reporter gene. Secreted APP also induced an increase in kappa B DNA-binding activity in hippocampal neurons treated with sAPP. Both effects were mimicked by an analog of cyclic GMP and inhibited by an antagonist of cyclic GMP-dependent protein kinase. Such activation of kappa B-dependent transcription was correlated in two ways with the ability of sAPP to protect neuronal cells against calcium-mediated damage: (1) tumor necrosis factor beta also protected against calcium-mediated insults and induced kappa B-dependent transcription; (2) antisense oligonucleotide-mediated reduction of an endogenous inhibitor of NF-kappa B activated kappa B-binding activity and attenuated calcium-mediated toxicity in both a neuronal cell line and in primary neurons. These findings suggest that a kappa B-binding transcription factor can act as a coordinator of neuroprotective gene expression in response to cytokines.

Amyloid beta-Protein Precursor↗
PubMed1996

Linoleic acid activates nuclear transcription factor-kappa B (NF-kappa B) and induces NF-kappa B-dependent transcription in cultured endothelial cells.

High dietary intakes of unsaturated fats may be atherogenic by disrupting normal functions of the vascular endothelium, due in part to the ability of linoleic acid (18:2n-6) to contribute to an increase in cellular oxidative stress and related injurious events. Exposing endothelial cells to 90 micromol linoleic acid/L for 6 h resulted in a significant increase in lipid hydroperoxides that coincided wih an increase in intracellular calcium concentrations. Treatment with this fatty acid caused an initial decrease in glutathione concentrations, which was followed by an increase at later time points. Most importantly, a significant activation of the oxidative stress-sensitive nuclear transcription factor-kappa B (NF-kappa B) was achieved after a 6-h exposure to 18:2n-6, which is the time point at which maximal depletion of cellular glutathione was observed. The fatty acid-mediated NF-kappa B activation was accompanied by induction of NF-kappa B-dependent transcription, as measured by chloramphenicol acetyltransferase (CAT) assay of an NF-kappa B-responsive promoter construct. Pretreatment of endothelial cells with vitamin E and N-acetyl cysteine inhibited the fatty acid-induced activation of NF-kappa B and formation of lipid hydroperoxides. These data suggest that oxidative stress-induced cellular changes are critical early events in fatty acid-mediated endothelial cell dysfunction.

Animals↗
PubMed1996

Linoleic acid and TNF-alpha cross-amplify oxidative injury and dysfunction of endothelial cells.

Factors implicated in the development of atherosclerosis include metabolic alterations of the endothelium induced by certain lipids and inflammatory cytokines. To study the hypothesis that the combined presence of unsaturated fatty acids and inflammatory cytokines may cross-amplify their individual injurious effects, cultured endothelial cells were treated with 90 mu M of linoleic acid (18:2 n-6) and/or 20 ng/ml (100 U/ml) of tumor necrosis factor-alpha (TNF) for up to 24 h. Disturbances in endothelial cell metabolism were determined by measuring cellular oxidative stress, oxidative stress-inducible nuclear factor-kappa B (NF-kappa B) and NF-kappa B-related transcription, intracellular calcium levels, and endothelial barrier function reflected by transendothelial albumin movement. Both 18:2 and TNF increased cellular oxidation, intracellular calcium, and endothelial barrier permeability. These changes were cross-amplified in cells treated both with 18:2 and TNF, compared with 18:2 or TNF alone. In contrast, a combined exposure to 18:2 and TNF did not potentiate effects mediated by 18:2 or TNF alone on NF-kappa B activation or NF-kappa B-related transcription. Pretreatment with 25 mu M vitamin E attenuated 18:2 and/or TNF-mediated endothelial cell dysfunction. These results suggest that certain unsaturated fatty acids can potentiate TNF-mediated endothelial cell dysfunction and that oxidative stress may be partially responsible for these metabolic events. These findings have implications for understanding lipid-mediated inflammatory responses in atherosclerosis.

Animals↗
PubMed1995

The secreted form of the Alzheimer's beta-amyloid precursor protein stimulates a membrane-associated guanylate cyclase.

We previously demonstrated that secreted forms of the Alzheimer's beta-amyloid precursor protein (sAPP) elevate cyclic GMP (cGMP) in primary neuronal cultures and that this effect is responsible for the modulation of neuronal calcium homoeostasis by sAPP. We have investigated further the mechanism by which sAPP elevates cGMP. Inhibition of the formation of nitric oxide or carbon monoxide did not affect the ability of sAPP to lower rapidly intraneuronal calcium levels or elevate cGMP, suggesting that sAPP does not activate a soluble (cytosolic) guanylate cyclase. A dose-dependent stimulation of cGMP formation by sAPP was observed in brain membrane preparations. The stimulation was also dependent on the presence of ATP. These data suggest that sAPP activates a membrane-associated guanylate cyclase, perhaps similar to those present in the receptors for the natriuretic peptides and sperm motility factors.

Adenosine Triphosphate↗
PubMed1995

Tumor necrosis factors alpha and beta protect neurons against amyloid beta-peptide toxicity: evidence for involvement of a kappa B-binding factor and attenuation of peroxide and Ca2+ accumulation.

In Alzheimer disease (AD) the amyloid beta-peptide (A beta) accumulates in plaques in the brain. A beta can be neurotoxic by a mechanism involving induction of reactive oxygen species (ROS) and elevation of intracellular free calcium levels ([Ca2+]i). In light of evidence for an inflammatory response in the brain in AD and reports of increased levels of tumor necrosis factor (TNF) in AD brain we tested the hypothesis that TNFs affect neuronal vulnerability to A beta. A beta-(25-35) and A beta-(1-40) induced neuronal degeneration in a concentration- and time-dependent manner. Pretreatment of cultures for 24 hr with TNF-beta or TNF-alpha resulted in significant attenuation of A beta-induced neuronal degeneration. Accumulation of peroxides induced in neurons by A beta was significantly attenuated in TNF-pretreated cultures, and TNFs protected neurons against iron toxicity, suggesting that TNFs induce antioxidant pathways. The [Ca2+]i response to glutamate (quantified by fura-2 imaging) was markedly potentiated in neurons exposed to A beta, and this action of A beta was suppressed in cultures pretreated with TNFs. Electrophoretic mobility-shift assays demonstrated an induction of a kappa beta-binding activity in hippocampal cells exposed to TNFs. Exposure of cultures to I kappa B (MAD3) antisense oligonucleotides, a manipulation designed to induce NF-kappa B, mimicked the protection by TNFs. These data suggest that TNFs protect hippocampal neurons against A beta toxicity by suppressing accumulation of ROS and Ca2+ and that kappa B-dependent transcription is sufficient to mediate these effects. A modulatory role for TNF in the neurodegenerative process in AD is proposed.

Amyloid beta-Peptides↗
PubMed1995

S100 beta protects hippocampal neurons from damage induced by glucose deprivation.

S100 beta is a calcium-binding protein elevated in Down's syndrome and Alzheimer's disease. Previous studies have demonstrated that S100 beta is trophic for several neuronal populations. We tested the influence of S100 beta on hippocampal neurons. The initial response included a rapid increase in [Ca2+]i similar to that elicited by S100 beta in other populations. S100 beta also substantially decreased cell death and loss of mitochondrial function resulting from glucose deprivation. Therefore, S100 beta exerts a neuroprotective influence on CNS neurons, suggesting that its elevation in neurological disorders may be a compensatory response.

Animals↗
PubMed1995

Exposure to polychlorinated biphenyls causes endothelial cell dysfunction.

Environmental chemicals, such as polychlorinated biphenyls (PCBs), may be atherogenic by disrupting normal functions of the vascular endothelium. To investigate this hypothesis, porcine pulmonary artery-derived endothelial cells were exposed to 3,3',4,4'-tetrachlorobiphenyl (PCB 77), 2,3,4,4',5-pentachlorobiphenyl (PCB 114), or 2,2',4,4',5,5'-hexachlorobiphenyl (PCB 153) for up to 24 hours. These PCBs were selected for their varying binding avidities with the aryl hydrocarbon (Ah) receptor and differences in their induction of cytochrome P450. PCB 77 and PCB 114 significantly disrupted, in a dose-dependent manner, endothelial barrier function by allowing an increase in albumin transfer across endothelial monolayers. These PCBs also contributed markedly to cellular oxidative stress, as measured by 2,7-dichlorofluorescin (DCF) fluorescence and lipid hydroperoxides, and caused a significant increase in intracellular calcium ([Ca2+]i) levels. Enhanced oxidative stress and [Ca2+]i in PCB 77- and PCB 114-treated cells were accompanied by increased activity and content of cytochrome P450 1A and by a decrease in the vitamin E content in the culture medium. In contrast to the effects of PCB 77 and PCB 114, cell exposure to PCB 153 had no effect on cellular oxidation, [Ca2+]i, or endothelial barrier function. These results suggest that certain PCBs may play a role in the development of atherosclerosis by causing endothelial cell dysfunction and a decrease in the barrier function of the vascular endothelium. It is possible that interaction of PCBs with the Ah receptor and activation of the cytochrome P450 1A subfamily are involved in this pathology.

Animals↗
PubMed1995

Role of glutathione redox cycle in TNF-alpha-mediated endothelial cell dysfunction.

Modulation of the glutathione redox cycle may influence tumor necrosis factor-alpha (TNF)-mediated disturbances of endothelial integrity. To test this hypothesis, normal endothelial cells or cells with either increased or decreased glutathione levels were exposed to 100 ng (500 U) TNF/ml. Increased glutathione levels were achieved by exposure to 0.2 mM N-acetyl-L-cysteine (NAC) and decreased glutathione levels by exposure to 25 microM buthionine sulfoximine (BSO). Several components of the glutathione redox cycle as well as markers of endothelial integrity, such as cytoplasmic free calcium and transendothelial albumin transfer, were measured in the treated cells. Exposure to TNF for 3 and 6 h decreased total glutathione levels, which was followed by an increase at later time points. Moreover, treatment with TNF resulted in an increase in the ratio of oxidized to reduced glutathione, intracellular free calcium, albumin transfer across endothelial monolayers and lipid hydroperoxides. However, an increase in lipid hydroperoxides was seen only when endothelial cell cultures were supplemented with iron. BSO treatment increased susceptibility of endothelial cells to TNF-mediated metabolic disturbances. On the other hand, NAC partially protected against TNF-induced injury to endothelial monolayers. Our results demonstrate the important role of the glutathione redox cycle in TNF-mediated disturbances of the vascular endothelium and indicate that modulation of glutathione levels may potentiate the injurious effects of this inflammatory cytokine.

Acetylcysteine↗
PubMed1995

Role of cyclic GMP in the regulation of neuronal calcium and survival by secreted forms of beta-amyloid precursor.

The Alzheimer's disease (AD) beta-amyloid precursor proteins (beta APPs) are large membrane-spanning proteins that give rise to the beta A4 peptide deposited in AD amyloid plaques. beta APPs can also yield soluble forms (APPss) that are potently neuroprotective against glucose deprivation and glutamate toxicity, perhaps through their ability to lower the intraneuronal calcium concentration ([Ca2+]i). We have investigated the mechanism through which APPss exert these effects on cultured hippocampal neurons. The ability of APPss to lower rapidly [Ca2+]i was mimicked by membrane-permeable analogues of cyclic AMP (cAMP) and cyclic GMP (cGMP), as well as agents that elevate endogenous levels of these cyclic nucleotides. However, only cGMP content was increased by APPs treatment, and specific inhibition of cGMP-dependent protein kinase (but not cAMP-dependent kinase) blocked the activity of APPss. A membrane-permeable analogue of cGMP (8-bromo-cGMP) also mimicked the ability of APPss to attenuate the elevation of [Ca2+]i by glutamate, apparently through inhibition of NMDA receptor activity. In addition, 8-bromo-cGMP afforded protection against glucose deprivation and glutamate toxicity, and the protection by APPss against glucose deprivation was blocked by an inhibitor of cGMP-dependent kinase. Together, these data suggest that APPss mediate their [Ca2+]i-lowering and excitoprotective effects on target neurons through increases in cGMP levels.

Alzheimer Disease↗
PubMed1994

Staurosporine, K-252a, and K-252b stabilize calcium homeostasis and promote survival of CNS neurons in the absence of glucose.

Staurosporine, K-252a, and the 9-carboxylic related compound K-252b are low-molecular-weight alkaloids from microbial origin that at high concentrations are kinase inhibitors and can antagonize the effects of neuronal growth factors. Paradoxically, we have found that very low concentrations of these agents (10 fM-10 nM) prolong the survival of hippocampal, septal, and cortical neurons deprived of glucose. These agents did not prevent the depletion of ATP caused by glucose deprivation. The large elevation of intracellular calcium levels that normally mediates glucose deprivation-induced damage was attenuated by staurosporine, K-252a, and K-252b. Western blot analysis using antiphosphotyrosine antibody showed that staurosporine and the K-252 compounds (10-100 pM) stimulated tyrosine phosphorylation of several different proteins. The tyrosine kinase inhibitor genistein significantly reduced the protective effect of staurosporine and the K-252 compounds, indicating that tyrosine phosphorylation was required for neuroprotection by these compounds. Taken together, the data demonstrate that low concentrations of staurosporine and the K-252 compounds can stabilize calcium homeostasis, possibly by a mechanism involving activation of receptor tyrosine kinase transduction pathways.

Alkaloids↗
PubMed1993

beta-Amyloid precursor protein mismetabolism and loss of calcium homeostasis in Alzheimer's disease.

The suspected involvement of the beta-amyloid precursor protein (beta APP) in the etiology of Alzheimer's disease (AD) has been strengthened by recent genetic evidence, but pursuit of the mechanisms involved will initially require basic cell biology approaches. Several studies have concentrated on toxic activities of beta-amyloid peptide (beta AP) itself, illuminating its contributions to excitotoxicity and calcium-mediated degeneration in general. We now know that generation of beta AP from beta APP also compromises the production of an important set of trophic factors: the secreted forms of beta APP (APPS), which may act--ironically--by conferring protection from calcium-mediated insults. Therefore, conditions which contribute to the formation of beta AP (possibly including ischemia) not only produce an agent which exacerbates calcium-mediated cell death, but also reduce the levels of one of the few factors able to rescue calcium homeostasis. The implications of these postulates and their relationship to the process of aging are discussed.

Aging↗
PubMed1993

beta-Amyloid precursor protein metabolites and loss of neuronal Ca2+ homeostasis in Alzheimer's disease.

Recent findings link altered processing of beta-amyloid precursor protein (beta APP) to disruption of neuronal Ca2+ homeostasis and an excitotoxic mechanism of cell death in Alzheimer's disease. A major pathway of beta APP metabolism results in the release of secreted forms of beta APP, APPss. These secreted forms are released in response to electrical activity and can modulate neuronal responses to glutamate, suggesting roles in developmental and synaptic plasticity. beta APP is upregulated in response to neural injury and APPss can protect neurons against excitotoxic or ischemic insults by stabilizing the intracellular Ca2+ concentration [Ca2+]i. An alternative beta APP processing pathway liberates intact beta-amyloid peptide, which can form aggregates that disrupt Ca2+ homeostasis and render neurons vulnerable to metabolic or excitotoxic insults. Genetic abnormalities (e.g. certain beta APP mutations or Down syndrome) and age-related changes in brain metabolism (e.g. reduced energy availability or increased oxidative stress) may favor accumulation of [Ca2+]i-destabilizing beta-amyloid peptide and diminish the release of [Ca2+]i-stabilizing, neuroprotective APPss.

Alzheimer Disease↗
PubMed1992

Disulfide-linked S100 beta dimers and signal transduction.

S100 beta is a calcium-binding protein with neurotrophic and mitogenic activities, both of which may be mediated by the protein's ability to stimulate an increase in intracellular free calcium ([Ca2+]i). These extracellular trophic activities of S100 beta require a disulfide-linked, dimeric form of the protein. In this chapter, we present a minireview on the current state of knowledge concerning extracellular functions of S100 beta, with emphasis on the potential relevance of these activities to neuropathological disorders. We also report a simplified procedure for preparation of pharmacological amounts of biologically active S100 beta dimers, based on the finding that formation of disulfide-linked S100 beta dimers can be stimulated by the presence of calcium or lipid.

Animals↗