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Amyloid Peptide (amyloid + peptide)
Selected AbstractsSelection of D -Amino-Acid Peptides That Bind to Alzheimer's Disease Amyloid Peptide A,1,42 by Mirror Image Phage DisplayCHEMBIOCHEM, Issue 8 2003Katja Wiesehan Dr. Abstract A mirror image phage display approach was used to identify novel and highly specific ligands for Alzheimer's disease amyloid peptide A,(1,42). A randomized 12-mer peptide library presented on M13 phages was screened for peptides with binding affinity for the mirror image of A,(1,42). After four rounds of selection and amplification the peptides were enriched with a dominating consensus sequence. The mirror image of the most representative peptide (D -pep) was shown to bind A,(1,42) with a dissociation constant in the submicromolar range. Furthermore, in brain tissue sections derived from patients that suffered from Alzheimer's disease, amyloid plaques and leptomeningeal vessels containing A, amyloid were stained specifically with a fluorescence-labeled derivative of D -pep. Fibrillar deposits derived from other amyloidosis were not labeled by D -pep. Possible applications of this novel and highly specific A, ligand in diagnosis and therapy of Alzheimer's disease are discussed. [source] Thiophilic interaction chromatography of Alzheimer's , -amyloid peptidesCHEMICAL BIOLOGY & DRUG DESIGN, Issue 2005S. Parry Abstract:, Alzheimer's disease is characterized by a progressive formation of senile plaques in the brain, the major constituent of which is , -amyloid (A,) peptide, a proteolytic product of the transmembrane , -amyloid precursor protein (APP). Prior to the measurement of levels of the A, peptide for diagnostic purposes, this peptide must be isolated from the myriad of proteins resident in the human serum. Thiophilic interaction chromatography is an effective method for the isolation of proteins and peptides containing clusters of aromatic residues such as tryptophan, phenylalanine and tyrosine. The purpose of the present study was to develop a protocol for binding and recovery of A, peptides (1,38), (1,40) and (1,42) to T-gels by varying T-gel type and elution conditions such as the salt concentration and type of eluent. We established the minimal salt concentration necessary for the binding of the A,(1,40) peptide to the 3S-gel; binding at that concentration was subsequently compared with that of model proteins, lysozyme and , -chymotrypsin and this methodology was extended to 2S-gels and PyS. , -Amyloid peptide (1,40) showed a remarkably strong affinity for all three types of T-gels in comparison to lysozyme and , -chymotrypsin and was found to bind best to 2S-gel. [source] CSF biomarker profile and diagnostic value in vascular dementiaEUROPEAN JOURNAL OF NEUROLOGY, Issue 2 2009G. P. Paraskevas Background and purpose:, The differential diagnosis between vascular dementia (VD) and Alzheimer's disease (AD) or mixed dementia (MD) is not always easy in clinical practice. The purpose of the present study was to evaluate the cerebrospinal fluid (CSF) biomarkers tau protein in its total (,T) or hyperphosphorylated at threonin-181(,P-181) form and beta amyloid peptide 1,42 (A,42) alone and their combinations to investigate their diagnostic value in the discrimination between VD and AD or MD. Methods:, The above CSF biomarkers were determined in duplicate and blind to the clinical diagnosis by double sandwich, enzyme-linked immunosorbent assay (ELISA) commercial kits (Innogenetics, Gent, Belgium) in 92 AD patients, 23 VD patients, 17 patients with MD and 68 controls. Results:, Alzheimer's disease and MD showed increased levels of ,T, ,P and reduced levels of A,42 as compared with the controls. The best discrimination between VD and AD or MD was achieved by the combination of all three biomarkers, correctly classifying ,85% of patients, either in the form of a discriminant function or in the form of the ,T × ,P-181/A,42 formula. Conclusions:, Cerebrospinal fluid biomarkers may be a useful adjunct for the discrimination between AD/ MD and VD in every day clinical practice. [source] Lipid-induced conformational transition of the amyloid core fragment A,(28,35) and its A30G and A30I mutantsFEBS JOURNAL, Issue 10 2008Sureshbabu Nagarajan The interaction of the ,-amyloid peptide (A,) with neuronal membranes could play a key role in the pathogenesis of Alzheimer's disease. Recent studies have focused on the interactions of A, oligomers to explain the neuronal toxicity accompanying Alzheimer's disease. In our study, we have investigated the role of lipid interactions with soluble A,(28,35) (wild-type) and its mutants A30G and A30I in their aggregation and conformational preferences. CD and Trp fluorescence spectroscopic studies indicated that, immediately on dissolution, these peptides adopted a random coil structure. Upon addition of negatively charged 1,2-dipalmitoyl- syn -glycero-3-phospho- rac -(glycerol) sodium salt (PG) lipid, the wild-type and A30I mutant underwent reorganization into a predominant ,-sheet structure. However, no conformational changes were observed in the A30G mutant on interaction with PG. In contrast, the presence of zwitterionic 1,2-dipalmitoyl- syn -glycero-3-phosphatidylcholine (PC) lipid had no effect on the conformation of these three peptides. These observations were also confirmed with atomic force microscopy and the thioflavin-T assay. In the presence of PG vesicles, both the wild-type and A30I mutant formed fibrillar structures within 2 days of incubation in NaCl/Pi, but not in their absence. Again, no oligomerization was observed with PC vesicles. The Trp studies also revealed that both ends of the three peptides are not buried deep in the vesicle membrane. Furthermore, fluorescence spectroscopy using the environment-sensitive probe 1,6-diphenyl-1,3,5-hexatriene showed an increase in the membrane fluidity upon exposure of the vesicles to the peptides. The latter effect may result from the lipid head group interactions with the peptides. Fluorescence resonance energy transfer experiments revealed that these peptides undergo a random coil-to-sheet conversion in solution on aging and that this process is accelerated by negatively charged lipid vesicles. These results indicate that aggregation depends on hydrophobicity and propensity to form ,-sheets of the amyloid peptide, and thus offer new insights into the mechanism of amyloid neurodegenerative disease. [source] Beta-amyloid peptide stimulates endozepine release in cultured rat astrocytes through activation of N -formyl peptide receptorsGLIA, Issue 13 2008Tursonjan Tokay Abstract Astroglial cells synthesize and release endozepines, a family of neuropeptides derived from diazepam-binding inhibitor (DBI). The authors have recently shown that ,-amyloid peptide (A,) stimulates DBI gene expression and endozepine release. The purpose of this study was to determine the mechanism of action of A, in cultured rat astrocytes. A,25,35 and the N -formyl peptide receptor (FPR) agonist N -formyl-Met-Leu-Phe (fMLF) increased the secretion of endozepines in a dose-dependent manner with EC50 value of ,2 ,M. The stimulatory effects of A,25,35 and the FPR agonists fMLF and N -formyl-Met-Met-Met (fMMM) on endozepine release were abrogated by the FPR antagonist N - t -Boc-Phe-Leu-Phe-Leu-Phe. In contrast, A,25,35 increased DBI mRNA expression through a FPR-independent mechanism. A,25,35 induced a transient stimulation of cAMP formation and a sustained activation of polyphosphoinositide turnover. The stimulatory effect of A,25,35 on endozepine release was blocked by the adenylyl cyclase inhibitor somatostatin, the protein kinase A (PKA) inhibitor H89, the phospholipase C inhibitor U73122, the protein kinase C (PKC) inhibitor chelerythrine and the ATP binding cassette transporter blocker glyburide. Taken together, these data demonstrate for the first time that A,25,35 stimulates endozepine release from rat astrocytes through a FPR receptor positively coupled to PKA and PKC. © 2008 Wiley-Liss, Inc. [source] Calcium/calmodulin-dependent protein kinase type IV is a target gene of the Wnt/,-catenin signaling pathway,JOURNAL OF CELLULAR PHYSIOLOGY, Issue 3 2009Macarena S. Arrázola Calcium/calmodulin-dependent protein kinase IV (CaMKIV) plays a key role in the regulation of calcium-dependent gene expression. The expression of CaMKIV and the activation of CREB regulated genes are involved in memory and neuronal survival. We report here that: (a) a bioinformatic analysis of 15,476 promoters of the human genome predicted several Wnt target genes, being CaMKIV a very interesting candidate; (b) CaMKIV promoter contains TCF/LEF transcription motifs similar to those present in Wnt target genes; (c) biochemical studies indicate that lithium and the canonical ligand Wnt-3a induce CaMKIV mRNA and protein expression levels in rat hippocampal neurons as well as CaMKIV promoter activity; (d) treatment of hippocampal neurons with Wnt-3a increases the binding of ,-catenin to the CaMKIV promoter: (e) In vivo activation of the Wnt signaling improve spatial memory impairment and restores the expression of CaMKIV in a mice double transgenic model for Alzheimer's disease which shows decreased levels of the kinase. We conclude that CaMKIV is regulated by the Wnt signaling pathway and that its expression could play a role in the neuroprotective function of the Wnt signaling against the Alzheimer's amyloid peptide. J. Cell. Physiol. 221: 658,667, 2009. © 2009 Wiley-Liss, Inc. [source] Imbalance of plasma membrane ion leak and pump relationship as a new aetiological basis of certain disease statesJOURNAL OF INTERNAL MEDICINE, Issue 6 2003G. Ronquist Abstract. The basis for life is the ability of the cell to maintain ion gradients across biological membranes. Such gradients are created by specific membrane-bound ion pumps [adenosine triphosphatases (ATPases)]. According to physicochemical rules passive forces equilibrate (dissipate) ion gradients. The cholesterol/phospholipid ratio of the membrane and the degree of saturation of phospholipid fatty acids are important factors for membrane molecular order and herewith a determinant of the degree of non-specific membrane leakiness. Other operative principles, i.e. specific ion channels can be opened and closed according to mechanisms that are specific to the cell. Certain compounds called ionophores can be integrated in the plasma membrane and permit specific inorganic ions to pass. Irrespective of which mechanism ions leak across the plasma membrane the homeostasis may be kept by increasing ion pumping (ATPase activity) in an attempt to restore the physiological ion gradient. The energy source for this work seems to be glycolytically derived ATP formation. Thus an increase in ion pumping is reflected by increased ATP hydrolysis and rate of glycolysis. This can be measured as an accumulation of breakdown products of ATP and end-products of anaerobic glycolysis (lactate). In certain disease entities, the balance between ATP formation and ion pumping may be disordered resulting in a decrease in inter alia (i.a.) cellular energy charge, and an increase in lactate formation and catabolites of adenylates. Cardiac syndrome X is proposed to be due to an excessive leakage of potassium ions, leading to electrocardiographic (ECG) changes, abnormal Tl-scintigraphy of the heart and anginal pain (induced by adenosine). Cocksackie B3 infections, a common agent in myocarditis might also induce an ionophore-like effect. Moreover, Alzheimer's disease is characterized by the formation of extracellular amyloid deposits in the brain of patients. Perturbation of cellular membranes by the amyloid peptide during the development of Alzheimer's disease is one of several mechanisms proposed to account for the toxicity of this peptide on neuronal membranes. We have studied the effects of the peptide and fragments thereof on 45Ca2+ -uptake in human erythrocytes and the energetic consequences. Treatment of erythrocytes with the ,1,40 peptide, results in qualitatively similar nucleotide pattern and decrease of energy charge as the treatment with Ca2+ -ionophore A23187. Finally, in recent studies we have revealed and published in this journal that a rare condition, Tarui's disease or glycogenosis type VII, primarily associated with a defect M-subunit of phosphofructokinase, demonstrates as a cophenomenon an increased leak of Ca2+ into erythrocytes. [source] Cell cycle-driven neuronal apoptosis specifically linked to amyloid peptide A,1,42 exposure is not exacerbated in a mouse model of presenilin-1 familial Alzheimer's diseaseJOURNAL OF NEUROCHEMISTRY, Issue 2 2008Bilal Malik Abstract We have shown previously that ,-catenin and cyclin D1 are up-regulated in cortical neurons from homozygous mice carrying the familial Alzheimer's disease (FAD) presenilin-1 M146V mutation in a knock-in model (PS1 KIM146V mice), leading to cell cycle-associated apoptosis. Here, we have aimed to determine (i) whether this phenotype is present in heterozygous PS1 KIM146V mice, which reflects more accurately the PS1 FAD condition in humans and (ii) whether A,1,42, which is invariably present in the PS1 FAD brain and is thought to affect neuronal cell cycle kinetics, may contribute to the abnormal cell cycle/cell death phenotype seen in PS1 KIM146V mice. We demonstrate that cell cycle-linked apoptosis occurs in heterozygous PS1 KIM146V post-mitotic neurons. In addition, there is a significant A,-associated increase in cell cycle and cell death that is not further modified by the PS1 KIM146V mutation. Our results are consistent with a cell cycle-associated neurodegeneration model in the PS1 FAD brain in which the loss of PS1-dependent ,-catenin regulatory function is sufficient to commit susceptible neurons to an abortive cell cycle, and may act synergistically with the A, cytotoxic challenge present in the PS1 FAD brain to expand the neuronal population susceptible to cell cycle-driven apoptosis. [source] Conventional protein kinase C isoforms mediate neuroprotection induced by phorbol ester and estrogenJOURNAL OF NEUROCHEMISTRY, Issue 1 2006Myriam Cordey Abstract Rapid signal transduction pathways play a prominent role in mediating neuroprotective actions of estrogen in the CNS. We have previously shown that estrogen-induced neuroprotection of primary cerebrocortical neurons from ,-amyloid peptide (A,) toxicity depends on activation of protein kinase C (PKC). PKC activation with phorbol-12-myristate-13-acetate (PMA) also provides neuroprotection in this paradigm. Because the PKC family includes several isoforms that have opposing roles in regulating cell survival, we sought to identify which PKC isoforms contribute to neuroprotection induced by PMA and estrogen. We detected protein expression of multiple PKC isoforms in primary neuron cultures, including conventional (,, ,I, ,II), novel (,, ,, ,) and atypical (,, ,/,) PKC. Using a panel of isoform-specific peptide inhibitors and activators, we find that novel and atypical PKC isoforms do not participate in the mechanism of either PMA or estrogen neuroprotection. In contrast, a selective peptide activator of conventional PKC isoforms provides dose-dependent neuroprotection against A, toxicity. In addition, peptide inhibitors of conventional, ,I, or ,II PKC isoforms significantly reduce protection afforded by PMA or 17,-estradiol. Taken together, these data provide evidence that conventional PKC isoforms mediate phorbol ester and estrogen neuroprotection of cultured neurons challenged by A, toxicity. [source] 22R -Hydroxycholesterol protects neuronal cells from ,-amyloid-induced cytotoxicity by binding to ,-amyloid peptideJOURNAL OF NEUROCHEMISTRY, Issue 5 2002Zhi-Xing Yao Abstract 22R -hydroxycholesterol, a steroid intermediate in the pathway of pregnenolone formation from cholesterol, was found at lower levels in Alzheimer's disease (AD) hippocampus and frontal cortex tissue specimens compared to age-matched controls. ,-Amyloid (A,) peptide has been shown to be neurotoxic and its presence in brain has been linked to AD pathology. 22R -hydroxycholesterol was found to protect, in a dose-dependent manner, against A,-induced rat sympathetic nerve pheochromocytoma (PC12) and differentiated human Ntera2/D1 teratocarcinoma (NT2N) neuron cell death. Other steroids tested were either inactive or acted on rodent neurons only. The effect of 22R -hydroxycholesterol was found to be stereospecific because its enantiomer 22S -hydroxycholesterol failed to protect the neurons from A,-induced cell death. Moreover, the effect of 22R -hydroxycholesterol was specific for A,-induced cell death because it did not protect against glutamate-induced neurotoxicity. The neuroprotective effect of 22R -hydroxycholesterol was seen when using A,1,42 but not the A,25,35 peptide. To investigate the mechanism of action of 22R -hydroxycholesterol we examined the direct binding of this steroid to A, using a novel cholesterol-protein binding blot assay. Using this method the direct specific binding, under native conditions, of 22R -hydroxycholesterol to A,1,42 and A,17,40, but not A,25,35, was observed. These data suggest that 22R -hydroxycholesterol binds to A, and the formed 22R -hydroxycholesterol/A, complex is not toxic to rodent and human neurons. We propose that 22R -hydroxycholesterol offers a new means of neuroprotection against A, toxicity by inactivating the peptide. [source] The small heat shock protein Hsp27 protects cortical neurons against the toxic effects of ,-amyloid peptideJOURNAL OF NEUROSCIENCE RESEARCH, Issue 14 2009Michael King Abstract Neurofibrillary tangles and amyloid plaques are considered to be hallmarks of Alzheimer's disease (AD), and the toxic effects of amyloid-, peptide (A,) lead to activation of stress-related signaling and neuronal loss. The small heat shock protein Hsp27 is reported to be increased in AD brains and to accumulate in plaques, but whether this represents a potentially protective response to stress or is part of the disease process is not known. We hypothesized that increased expression of Hsp27 in neurons can promote neuronal survival and stabilize the cytoskeleton in the face of A, exposure. By using neonatal rat cortical neurons, we investigated the potential role of Hsp27 in neuronal cultures in the presence or absence of A,. We initially tested whether a heat stress (HS) would be sufficient to induce endogenous Hsp27 expression. HS not only did not result in neuronal Hsp27 up-regulation but made the cells more vulnerable to A, exposure. We then used cDNA transfection to overexpress EGFP-Hsp27 (or the empty vector) in cultures and then assessed neuronal survival and growth. Transfected neurons appeared healthy and had robust neuritic outgrowth. A, treatment induced significant cell death by 48,72 hr in nontransfected and empty-vector-expressing cultures. In contrast, cultures expressing Hsp27 did not display significant apoptosis. Our results show that Hsp27-expressing neurons were selectively protected against the deleterious effects of A, treatment; neuronal degeneration was prevented, and A,-induced alterations in mitochondrial size were attenuated. We also demonstrate that Hsp27 expression can enhance neurite growth in cortical neurons compared with control vector-transfected cells. Overall, our study provides new evidence that Hsp27 can provide a protective influence in primary cortical neurons in the face of toxic concentrations of amyloid. © 2009 Wiley-Liss, Inc. [source] Lack of neprilysin suffices to generate murine amyloid-like deposits in the brain and behavioral deficit in vivoJOURNAL OF NEUROSCIENCE RESEARCH, Issue 8 2006Rime Madani Abstract Accumulation of the ,-amyloid peptide (A,) in the brain is a major pathological hallmark of Alzheimer's disease (AD), leading to synaptic dysfunction, neuronal death, and memory impairment. The levels of neprilysin, a major A,-degrading enzyme, are decreased in AD brains and during aging. Because neprilysin cleaves A, in vivo, its down-regulation may contribute to the pathophysiology of AD. The aim of this study was to assess the consequences of neprilysin deficiency on accumulation of murine A, in brains and associated pathologies in vivo by investigating neprilysin-deficient mice on biochemical, morphological, and behavioral levels. Aged neprilysin-deficient mice expressed physiological amyloid precursor protein (APP) levels and exhibited elevated brain A, concentrations and amyloid-like deposits in addition to signs of neuronal degeneration in their brains. Behaviorally, neprilysin-deficient mice acquired a significantly weaker conditioned taste aversion that extinguished faster than the aversion of age-matched controls. Our data establish that, under physiological APP expression levels, neprilysin deficiency is associated with increased A, accumulation in the brain and leads to deposition of amyloid-like structures in vivo as well as with signs of AD-like pathology and with behavioral deficits. © 2006 Wiley-Liss, Inc. [source] Antibodies against ,-amyloid reduce a, oligomers, glycogen synthase kinase-3, activation and , phosphorylation in vivo and in vitroJOURNAL OF NEUROSCIENCE RESEARCH, Issue 3 2006Qiu-Lan Ma Abstract Although active and passive immunization against the ,-amyloid peptide (A,) of amyloid plaque-bearing transgenic mice markedly reduces amyloid plaque deposition and improves cognition, the mechanisms of neuroprotection and impact on toxic oligomer species are not understood. We demonstrate that compared to control IgG2b, passive immunization with intracerebroventricular (icv) anti-A, (1,15) antibody into the AD HuAPPsw (Tg2576) transgenic mouse model reduced specific oligomeric forms of A,, including the dodecamers that correlate with cognitive decline. Interestingly, the reduction of soluble A, oligomers, but not insoluble A,, significantly correlated with reduced , phosphorylation by glycogen synthase kinase-3, (GSK-3,), a major , kinase implicated previously in mediating A, toxicity. A conformationally-directed antibody against amyloid oligomers (larger than tetramer) also reduced A, oligomer-induced activation of GSK3, and protected human neuronal SH-SY5Y cells from A, oligomer-induced neurotoxicity, supporting a role for A, oligomers in human , kinase activation. These data suggest that antibodies that are highly specific for toxic oligomer subspecies may reduce toxicity via reduction of GSK-3,, which could be an important strategy for Alzheimer's disease (AD) therapeutics. © 2005 Wiley-Liss, Inc. [source] Caspase cleavage of exon 9 deleted presenilin-1 is an early event in apoptosis induced by calcium ionophore A 23187 in SH-SY5Y neuroblastoma cellsJOURNAL OF NEUROSCIENCE RESEARCH, Issue 1 2001Bogdan O. Popescu Abstract Presenilins (PSs) are mutated in a majority of familial Alzheimer disease (FAD) cases. Mutated PSs may cause FAD by a number of pro-apoptotic mechanisms, or by regulating ,-secretase activity, a protease involved in ,-amyloid precursor protein processing to the neurotoxic ,-amyloid peptide. Besides their normal endoproteolytic processing, PSs are substrates for caspases, being cleaved to alternative N-terminal and C-terminal fragments. So far little is known about the role of PSs cleavage in the apoptotic machinery. Here, we used SH-SY5Y neuroblastoma cells stably transfected with wild-type or exon 9 deleted presenilin 1 (PS1) in a time-course study after the exposure to the calcium ionophore A23187. During and after exposure to A 23187, intracellular calcium levels were higher in exon 9 deleted PS1 cells as compared with non-transfected and wild-type PS1 transfected cells. Cell death and the enrichment of apoptotic cells after A23187 exposure were increased by overexpression of exon 9 deleted PS1 as compared with the control cell lines. Wild-type PS1 cells were compared with exon 9 deleted PS1 cells and the temporal relationship between PS1 and other caspase substrates cleavages was analyzed. Exon 9 deleted PS1 cells exhibited a higher caspase-3 activation and a greater cleavage of PS1 and poly(ADP-ribose) polymerase (PARP) compared with wild-type PS1 cells. Exon 9 deleted PS1 cleavage occurred earlier than other caspase substrate cleavages (i.e., PARP and gelsolin), simultaneous with minimum detectable caspase-3 activation. Therefore, alternative cleavage of PS1 may play an important role for the regulation of the proteolytic cascade activated during apoptosis. J. Neurosci. Res. 66:122,134, 2001. © 2001 Wiley-Liss, Inc. [source] Synthesis of A,[1-42] and its derivatives with improved efficiencyJOURNAL OF PEPTIDE SCIENCE, Issue 2 2007Márta Zarándi Abstract It has been proved that the principal component of senile plaques is aggregates of ,-amyloid peptide (A,) in cases of one of the most common forms of age-related neurodegenerative disorders, Alzheimer's disease (AD). Although the synthetic methods for the synthesis of A, peptides have been developed since their first syntheses, A,[1-42] is still problematic to prepare. The highly hydrophobic composition of A,[1-42] results in aggregation between resin-bound peptide chains or intrachain aggregation which leads to a decrease in the rates of deprotection and repetitive incomplete coupling reactions during 9-flurenylmethoxycarbonyl (Fmoc) synthesis. In order to avoid aggregation and/or disrupt internal aggregation during stepwise Fmoc solid phase synthesis and to improve the quality of crude products, several attempts have been made. Since highly pure A, peptides in large quantities are used in biological experiments, we wanted to develop a method for a rational synthesis of human A,[1-42] with high purity and adequate yield. This paper reports a convenient methodology with a novel solvent system for the synthesis of A,[1-42], its N -terminally truncated derivatives A,[4-42] and A,[5-42], and A,[1-42] labeled with 7-amino-4-methyl-3-coumarinylacetic acid (AMCA) at the N -terminus using Fmoc strategy. The use of 10% anisole in Dimethylformamide/Dichloromethane (DMF/DCM) can substantially improve the purity and yield of crude A,[1-42] and has been shown to be an optimal coupling condition for the synthesis of A,[1-42]. Anisole is a cheap and simple aid in the synthesis of ,difficult sequences' where other solvents are less successful in the prevention of aggregation during the synthesis. Copyright © 2006 European Peptide Society and John Wiley & Sons, Ltd. [source] Cerebrospinal fluid brain-derived proteins in the diagnosis of Alzheimer's disease and Creutzfeldt,Jakob diseaseNEUROPATHOLOGY & APPLIED NEUROBIOLOGY, Issue 6 2002A. J. E. GreenArticle first published online: 24 NOV 200 The differential diagnosis of dementia can be difficult in the early stages of disease, and with the emergence of new therapeutic agents for Alzheimer's disease (AD) there is an increasing need for reliable and accurate diagnostic tests. The concept of brain-specific proteins was first proposed in the 1960s and, since that time, methods have developed to measure these proteins in the cerebrospinal fluid (CSF). The concentration of individual brain-specific proteins can be altered in disease, and these changes are thought to reflect the underlying pathology. CSF tau protein and amyloid peptide A,42 concentrations are altered in AD and have been proposed as early diagnostic tests for this disease. The data from a number of studies suggest that these proteins may be of value, but are less specific than previously thought and further studies with neuropathological confirmation are required before these tests can be introduced into clinical practice. The detection of 14-3-3 in the CSF is an accurate test for sporadic Creutzfeldt,Jakob disease (CJD) and this accuracy has lead the World Health Organization to revise the clinical criteria for probable sporadic CJD to include a positive CSF 14-3-3. However, CSF 14-3-3 is less useful in the diagnosis of variant CJD, where studies are underway investigating the value of other CSF proteins. [source] Mutagenic analysis of the nucleation propensity of oxidized Alzheimer's ,-amyloid peptidePROTEIN SCIENCE, Issue 8 2005Tony Christopeit Abstract The formation of polypeptide aggregates represents a nucleated polymerization reaction in which an initial nucleation event (lag phase) is followed by the extension of newly formed nuclei into larger aggregates, including fibrils (growth phase). The efficiencies of these reactions relate to the lag time (lag phase) and to the rate of aggregation (growth phase), which can be determined from experimental aggregation curves. Here we present a mutagenic analysis in which we replace valine 18 of the Alzheimer's A, (1,40) peptide with 17 different amino acids and determine its effect on the lag time, and therefore, on the propensity of nucleation. Comparison with various physico-chemical properties shows that nucleation is affected in a predictable manner depending on the ,-sheet propensity and hydrophobicity of residue 18. In addition, we observe a direct proportionality between the lag time and the rate of aggregation. These data imply that the two reactions, nucleation and polymerization, are governed by very similar physicochemical principles and that they involve the formation of the same types of noncovalent interactions. [source] Cathepsin protease activity modulates amyloid load in extracerebral amyloidosisTHE JOURNAL OF PATHOLOGY, Issue 4 2006C Röcken Abstract In cerebral amyloidoses, such as Alzheimer's disease, proteolytic processing of the precursor protein is a fundamental mechanism of the disease, since it generates the amyloid protein. However, the putative significance of proteases in extracerebral amyloidoses is less well defined. In this study, we investigated the biological significance of cathepsin (Cath) B, CathK, and CathL in the pathology and pathogenesis of extracerebral amyloidoses by using the murine model of reactive or secondary AA amyloidosis with three different cathepsin-deficient mouse strains. Extracerebral AA amyloid was induced by injecting amyloid-enhancing factor and silver nitrate into CathB,/,, CathK,/,, and CathL,/, mice. Wild-type mice served as a control. CathK,/, mice deposited over 90% more amyloid and CathL,/, mice 60% less amyloid than the control (p < 0.0001). The amyloid load in CathB,/, mice did not differ from that in wild-type mice. In vitro degradation experiments with recombinant human and murine serum amyloid A (SAA) 1.1 and CathK and CathL showed that CathL generates a large number of differently sized SAA cleavage products. One of these fragments spans the heparin/heparan sulphate binding site and the neutral cholesterol ester hydrolase activating region of SAA. CathK showed only endoproteolytic activity and did not generate any AA amyloid-like peptides. This study provides unequivocal evidence that proteases modulate amyloid load in extracerebral amyloidosis. CathL was identified as an amyloid-promoting and CathK as an amyloid-retarding cysteine protease. CathB may only modulate the primary structure of the amyloid peptide without affecting amyloid load. Copyright © 2006 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd. [source] Simvastatin therapy prevents brain trauma-induced increases in ,-amyloid peptide levels,ANNALS OF NEUROLOGY, Issue 3 2009Eric E. Abrahamson PhD Elevations in ,-amyloid peptide (A,) levels after traumatic brain injury (TBI) may confer risk for developing Alzheimer's disease in head trauma patients. We investigated the effects of simvastatin, a 3-hydroxy-3-methylglutaryl-CoA reductase inhibitor, on hippocampal A, burden in a clinically relevant head injury/intervention model using mice expressing human A,. Simvastatin therapy blunted TBI-induced increases in A,, reduced hippocampal tissue damage and microglial activation, and improved behavioral outcome. The ability of statins to reduce post-injury A, load and ameliorate pathological sequelae of brain injury makes them potentially effective in reducing the risk of developing Alzheimer's disease in TBI patients. Ann Neurol 2009;66:407,414 [source] Key Factors in Alzheimer's Disease: ,-amyloid Precursor Protein Processing, Metabolism and Intraneuronal TransportBRAIN PATHOLOGY, Issue 1 2001Thomas A. Bayer During the last years it has become evident that the ,-amyloid (A,) component of senile plaques may be the key molecule in the pathology of Alzheimer's disease (AD). The source and place of the neurotoxic action of A,, however, is still a matter of controversy. The precursor of the ,-amyloid peptide is the predominantly neuronal ,-amyloid precursor protein. We, and others, hypothesize that intraneuronal misregulation of APP leads to an accumulation of A, peptides in intracellular compartments. This accumulation impairs APP trafficking, which starts a cascade of pathological changes and causes the pyramidal neurons to degenerate. Enhanced A, secretion as a function of stressed neurons and remnants of degenerated neurons provide seeds for extracellular A, aggregates, which induce secondary degenerative events involving neighboring cells such as neurons, astroglia and macrophages/microglia. [source] Promotion of axonal maturation and prevention of memory loss in mice by extracts of Astragalus mongholicusBRITISH JOURNAL OF PHARMACOLOGY, Issue 5 2006C Tohda Background and purpose: Neurons with atrophic neurites may remain alive and therefore may have the potential to regenerate even when neuronal death has occurred in some parts of the brain. This study aimed to explore effects of drugs that can facilitate the regeneration of neurites and the reconstruction of synapses even in severely damaged neurons. Experimental approach: We investigated the effects of extracts of Astragalus mongholicus on the cognitive defect in mice caused by injection with the amyloid peptide A,(25-35). We also examined the effect of the extract on the regeneration of neurites and the reconstruction of synapses in cultured neurons damaged by A,(25-35). Key results: A. mongholicus extract (1 g kg,1 day,1 for 15 days, p.o.) reversed A,(25-35)-induced memory loss and prevented the loss of axons and synapses in the cerebral cortex and hippocampus in mice. Treatment with A,(25-35) (10 ,M) induced axonal atrophy and synaptic loss in cultured rat cortical neurons. Subsequent treatment with A. mongholicus extract (100 ,g/ml) resulted in significant axonal regeneration, reconstruction of neuronal synapses, and prevention of A,(25-35)-induced neuronal death. Similar extracts of A. membranaceus had no effect on axonal atrophy, synaptic loss, or neuronal death. The major known components of the extracts (astragalosides I, II, and IV) reduced neurodegeneration, but the activity of the extracts did not correlate with their content of these three astragalosides. Conclusion and implications: A. mongholicus is an important candidate for the treatment of memory disorders and the main active constituents may not be the known astragalosides. British Journal of Pharmacology (2006) 149, 532,541. doi:10.1038/sj.bjp.0706865 [source] Selection of D -Amino-Acid Peptides That Bind to Alzheimer's Disease Amyloid Peptide A,1,42 by Mirror Image Phage DisplayCHEMBIOCHEM, Issue 8 2003Katja Wiesehan Dr. Abstract A mirror image phage display approach was used to identify novel and highly specific ligands for Alzheimer's disease amyloid peptide A,(1,42). A randomized 12-mer peptide library presented on M13 phages was screened for peptides with binding affinity for the mirror image of A,(1,42). After four rounds of selection and amplification the peptides were enriched with a dominating consensus sequence. The mirror image of the most representative peptide (D -pep) was shown to bind A,(1,42) with a dissociation constant in the submicromolar range. Furthermore, in brain tissue sections derived from patients that suffered from Alzheimer's disease, amyloid plaques and leptomeningeal vessels containing A, amyloid were stained specifically with a fluorescence-labeled derivative of D -pep. Fibrillar deposits derived from other amyloidosis were not labeled by D -pep. Possible applications of this novel and highly specific A, ligand in diagnosis and therapy of Alzheimer's disease are discussed. [source] Tacrine,Melatonin Hybrids as Multifunctional Agents for Alzheimer's Disease, with Cholinergic, Antioxidant, and Neuroprotective PropertiesCHEMMEDCHEM, Issue 5 2009María Isabel Fernández-Bachiller Dr. Abstract Tacrine,melatonin hybrids are potential multifunctional drugs for Alzheimer's disease that may simultaneously palliate intellectual deficits and protect the brain against both ,-amyloid peptide and oxidative stress. Molecular modeling studies show that they target both the catalytic active site (CAS) and the peripheral anionic site (PAS) of AChE. They are nontoxic and may be able to penetrate the CNS, according to in,vitro PAMPA-BBB assays. Tacrine,melatonin hybrids were designed and synthesized as new multifunctional drug candidates for Alzheimer's disease. These compounds may simultaneously palliate intellectual deficits and protect the brain against both ,-amyloid (A,) peptide and oxidative stress. They show improved cholinergic and antioxidant properties, and are more potent and selective inhibitors of human acetylcholinesterase (hAChE) than tacrine. They also capture free radicals better than melatonin. Molecular modeling studies show that these hybrids target both the catalytic active site (CAS) and the peripheral anionic site (PAS) of AChE. At sub-micromolar concentrations they efficiently displace the binding of propidium iodide from the PAS and could thus inhibit A, peptide aggregation promoted by AChE. Moreover, they also inhibit A, self-aggregation and display neuroprotective properties in a human neuroblastoma line against cell death induced by various toxic insults, such as A,25,35, H2O2, and rotenone. Finally, they exhibit low toxicity and may be able to penetrate the central nervous system according to an in,vitro parallel artificial membrane permeability assay for the blood,brain barrier (PAMPA-BBB). [source] Differential physiologic responses of ,7 nicotinic acetylcholine receptors to ,-amyloid1,40 and ,-amyloid1,42DEVELOPMENTAL NEUROBIOLOGY, Issue 1 2003Daniel H.S. Lee Abstract The ,-amyloid peptides (A,), A,1,40 and A,1,42, have been implicated in Alzheimer's disease (AD) pathology. Although A,1,42 is generally considered to be the pathological peptide in AD, both A,1,40 and A,1,42 have been used in a variety of experimental models without discrimination. Here we show that monomeric or oligomeric forms of the two A, peptides, when interact with the neuronal cation channel, ,7 nicotinic acetylcholine receptors (,7nAChR), would result in distinct physiologic responses as measured by acetylcholine release and calcium influx experiments. While A,1,42 effectively attenuated these ,7nAChR-dependent physiology to an extent that was apparently irreversible, A,1,40 showed a lower inhibitory activity that could be restored upon washings with physiologic buffers or treatment with ,7nAChR antagonists. Our data suggest a clear pharmacological distinction between A,1,40 and A,1,42. © 2003 Wiley Periodicals, Inc. J Neurobiol 55: 25,30, 2003 [source] Human brain carboxypeptidase B, which cleaves ,-amyloid peptides in vitro, is expressed in the endoplasmic reticulum of neuronsEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 9 2001Akira Matsumoto Abstract Intracellular localization of novel human brain carboxypeptidase B (HBCPB) was investigated in human hippocampus, using immunohistochemistry by confocal laser microscopy and biochemical purification of the homogenate by density gradient ultracentrifugation. The former revealed that the majority of HBCPB was expressed in the endoplasmic reticulum, in which the HBCPB-specific C14-module immunoreactivity was colocalized with GRP78 immunoreactivity, a stress 70 heat shock protein specifically expressed in the endoplasmic reticulum. The latter showed that anti-C14-module immunoreactivity and prepro-HBCPB immunoreactivity were both enriched in the microsome fraction, especially in that of the endoplasmic reticulum-density fraction of normal human hippocampal homogenates from various sources. However, HBCPB prepared from human hippocampus showed exopeptidase activity for synthetic ,-amyloid 1,42 peptide, in which A, X-42 C-terminus immunoreactivity was decreased in a fashion dose-dependent of the amount of the protease added. These findings indicate that HBCPB, which is expressed in the endoplasmic reticulum of a group of neuronal perikarya, may play an important physiological role in degradation of ,-amyloid 1,42, which is specifically generated in the endoplasmic reticulum of human and rodent neurons and is also regarded as the most pathogenic and aggregatable species among all ,-amyloid peptides. [source] Transplanted astrocytes internalize deposited ,-amyloid peptides in a transgenic mouse model of Alzheimer's diseaseGLIA, Issue 2 2008Rea Pihlaja Abstract Alzheimer's disease (AD) is one of the most devastating neurodegenerative disorders. The neuropathological hallmarks include extracellular senile plaques consisting of deposited ,-amyloid (A,) peptides and intraneuronal neurofibrillary tangles. Neuroinflammation and activation of astrocytes are also well-established features of AD neuropathology; however, the relationships between astrocytes and A, deposition remain unclear. Previous studies have shown that adult mouse astrocytes internalize and degrade A, deposits in brain sections prepared from human amyloid precursor protein (APP) transgenic mice. In the present study, we demonstrate that cultured adult, but not neonatal mouse astrocytes, respond morphologically and degrade A, deposits present in human AD brain. We also transplanted astrocytes isolated from enhanced green fluorescent protein expressing adult and neonatal mice into the hippocampi of human A, plaque-bearing transgenic APPSwe+PS1dE9 (APdE9) mice and their wild-type littermates and followed the migration and localization of these astrocytes by confocal microscopy upto 7 days after transplantation. Posttransplantation the astrocytes localized as aggregates or thin strings of many cells within the hippocampi of APdE9 and wild-type mice and showed limited migration from the injection site. Interestingly, most of the transplanted astrocytes were found near A, deposits in the hippocampi of APdE9 mice. In contrast to findings in ex vivo degradation assay, confocal microscopy revealed that both adult and neonatal transplanted astrocytes internalized human A, immunoreactive material in vivo. These results support the role of astrocytes as active A, clearing cells in the CNS that may have important implications for future development of therapeutic strategies for AD. © 2007 Wiley-Liss, Inc. [source] Effects of the monomeric, oligomeric, and fibrillar A,42 peptides on the proliferation and differentiation of adult neural stem cells from subventricular zoneJOURNAL OF NEUROCHEMISTRY, Issue 2 2007Chaejeong Heo Abstract The incidence of amyloid plaques, composed mainly of ,-amyloid peptides (A,), does not correlate well with the severity of neurodegeneration in patients with Alzheimer's disease (AD). The effects of A,42 on neurons or neural stem cells (NSCs) in terms of the aggregated form remain controversial. We prepared three forms of oligomeric, fibrillar, and monomeric A,42 peptides and investigated their effects on the proliferation and neural differentiation of adult NSCs, according to the degree of aggregation or concentration. A low micromolar concentration (1 ,mol/L) of oligomeric A,42 increased the proliferation of adult NSCs remarkably in a neurosphere assay. It also enhanced the neuronal differentiation of adult NSCs and their ability to migrate. These results provide us with valuable information regarding the effects of A,42 on NSCs in the brains of patients with AD. [source] Advanced glycation endproducts and pro-inflammatory cytokines in transgenic Tg2576 mice with amyloid plaque pathologyJOURNAL OF NEUROCHEMISTRY, Issue 2 2003Gerald Münch Abstract Increased expression and altered processing of the amyloid precursor protein (APP) and generation of ,-amyloid peptides is important in the pathogenesis of amyloid plaques in Alzheimer's disease (AD). Transgenic Tg2576 mice overexpressing the Swedish mutation of human APP exhibit ,-amyloid deposition in the neocortex and limbic areas, accompanied by gliosis and dystrophic neurites. However, murine plaques appear to be less cross-linked and the mice show a lower degree of inflammation and neurodegeneration than AD patients. ,Advanced glycation endproducts (AGEs)', formed by reaction of proteins with reactive sugars or dicarbonyl compounds, are able to cross-link proteins and to activate glial cells, and are thus contributing to plaque stability and plaque-induced inflammation in AD. In this study, we analyze the tissue distribution of AGEs and the pro-inflammatory cytokines IL-1, and TNF-, in 24-month-old Tg2576 mice, and compare the AGE distribution in these mice with a younger age group (13 months old) and a typical Alzheimer's disease patient. Around 70% of the amyloid plaque cores in the 24-month-old mice are devoid of AGEs, which might explain their solubility in physiological buffers. Plaque associated glia, which express IL-1, and TNF-,, contain a significant amount of AGEs, suggesting that plaques, i.e. A, as its major component, can induce intracellular AGE formation and the expression of the cytokines on its own. In the 13-month-old transgenic mice, AGEs staining can neither be detected in plaques nor in glial cells. In contrast, AGEs are present in high amounts in both plaques and glia in the human AD patient. The data obtained in this show interesting differences between the transgenic mouse model and AD patients, which should be considered using the transgenic approach to test therapeutical strategies to eliminate plaques or to attenuate the inflammatory response in AD. [source] Presenilin 1 is involved in the maturation of ,-site amyloid precursor protein-cleaving enzyme 1 (BACE1)JOURNAL OF NEUROSCIENCE RESEARCH, Issue 1 2007Akira Kuzuya Abstract One of the pathologic hallmarks of Alzheimer's disease is the excessive deposition of ,-amyloid peptides (A,) in senile plaques. A, is generated when ,-amyloid precursor protein (APP) is cleaved sequentially by ,-secretase, identified as ,-site APP-cleaving enzyme 1 (BACE1), and ,-secretase, a putative enzymatic complex containing presenilin 1 (PS1). However, functional interaction between PS1 and BACE1 has never been known. In addition to this classical role in the generation of A, peptides, it has also been proposed that PS1 affects the intracellular trafficking and maturation of selected membrane proteins. We show that the levels of exogenous and endogenous mature BACE1 expressed in presenilin-deficient mouse embryonic fibroblasts (PS,/,MEFs) were reduced significantly compared to those in wild-type MEFs. Moreover, the levels of mature BACE1 were increased in human neuroblastoma cell line, SH-SY5Y, stably expressing wild-type PS1, compared to native cells. Conversely, the maturation of BACE1 was compromised under the stable expression of dominant,negative mutant PS1 overexpression. Immunoprecipitation assay showed that PS1 preferably interacts with proBACE1 rather than mature BACE1, indicating that PS1 can be directly involved in the maturation process of BACE1. Further, endogenous PS1 was immunoprecipitated with endogenous BACE1 in SH-SY5Y cells and mouse brain tissue. We conclude that PS1 is directly involved in the maturation of BACE1, thus possibly functioning as a regulator of both ,- and ,-secretase in A, generation. © 2006 Wiley-Liss, Inc. [source] Pyruvate protection against ,-amyloid-induced neuronal death: Role of mitochondrial redox stateJOURNAL OF NEUROSCIENCE RESEARCH, Issue 2 2003Gema Alvarez Abstract The mechanism by which ,-amyloid protein (A,) causes degeneration in cultured neurons is not completely understood, but several lines of evidence suggest that A,-mediated neuronal death is associated with an enhanced production of reactive oxygen species (ROS) and oxidative damage. In the present study, we address whether supplementation of glucose-containing culture media with energy substrates, pyruvate plus malate (P/M), protects rat primary neurons from A,-induced degeneration and death. We found that P/M addition attenuated cell death evoked by ,-amyloid peptides (A,25,35 and A,1,40) after 24 hr treatment and that this effect was blocked by ,-ciano-3-hydroxycinnamate (CIN), suggesting that it requires mitochondrial pyruvate uptake. P/M supply to control and A,-treated neuronal cultures increases cellular reducing power, as indicated by the ability to reduce the dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The early increases in ROS levels, measured by dichlorofluorescein (DCF) fluorescence, and caspase-3 activity that follow exposure to A, were notably reduced in the presence of P/M. These results place activation of caspase-3 most likely downstream of oxidative damage to the mitochondria and indicate that mitochondrial NAD(P) redox status plays a central role in the neuroprotective effect of pyruvate. Inhibition of respiratory chain complexes and mitochondrial uncoupling did not block the early increase in ROS levels, suggesting that A, could initiate oxidative stress by activating a source of ROS that is not accesible to the antioxidant defenses fueled by mitochondrial substrates. © 2003 Wiley-Liss, Inc. [source] | ||||||||||||||||