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Apoptosis and messenger RNA in motor trigeminal neurons in ALS
9thMectingoftheBSN
5
RXRRES8ION OF NQF AND BDNF IN AN AllHlAL YODlcL Or CSRONIC NXURONAL F-IN ~IZONIRTRBBRSAL M.K.Brikkner,
s.Pagliusi (*) and Th.Arendt Paul Flechsig Inst. Brain Dept. Neurochem., Research, University Leipzig, Germany and (*) Glaxo Inst. Molec. Biol. S.A., Geneva, Switzerland Neuronal injury and cell death in the basal nucleus both in Alzheimer's disease and after chronic intake of ethanol in human and rat appears to be associated with a plastic adaptive rasponse resulting in a remodelling of the dendritic organization. This process which is associated with both degeneration of dendrites and sprouting of new dendritic elements is probably mediated by trophic signals derived from afferent and efferent neuronal connections and from neuroglia. In the precompared the sent study, we have therefore pattern of neuronal degeneration and reorganization in the basal forebrain of rat brain induced by chronic treatment with ethanol with the pattern of the expression of nerve growth factor (NGF) and brain derived growth factor (BDNF) obtained in the basal nucleus and its target regions such as neocortex and hippocampus by means of mRNA northern blot technique and in situ hybridisation.
B2
6 MEASURING INOSITOL PHOSPHATES AND CDP-DIACYLGLYCEROL REVEALS DIFFERENT MECHANISMS FOR RECEPTOR ACTIVATION OF PHOSPHOLIPASE C E. Claro1r2, John N. Fai;', F. Picatiste' Departments of Biochemistry, University of Tennessee, Memphis, USA~, and Universidad Aut&oma de Barcelona, Spain2 Carbachol (CCH), noradrenaline (NA) and ionomycin (ION) Stimulated accumulation of inositol phosphates (InsP's) and CDP-diacylglycerol (CDP-DAG) were studied in rat corti_ cal slices. Relative to maximally effective concentrations of CCH (ImM), the InsP's responses due to NA (3OuN) and ION (3uM) were 231% and 3256, respectively, whereas the effi cacies of NA and ION towards the CDP-DAG response were 63% and 29% of that of CCH. Both InsP's and CDP-DAG are produced upon activation of phospholipase C (PLC), but the latter is formed after diacylglycerol in a rea'ction which is inhibited by calcium. Also, in contrast to CCH, NA is unable to stimulate PLC in cortical membranes in the presence of GTP(S) . These results suggest that, unlike muscarinic receptors, which are directly coupled to PLC through a G protein, alpha, adrenergic receptors stimulate PLC indirectly most probably by raising intracellular Ca2+ concentration.
8
7 ~OP~sIS
MOTOR
AI00 )(IcSSRNGER RNA IN TRIGE==NRWRONS INALS.
11.A.8~rtIMnn', S. Ton&c*' and D.Y. Sun'. ‘Dapaarmm+ of Pathology, University of Wisconrin,
Madison, WI, 53706 'RockefellerUniversity, New York, 10021, USA
N.Y.
In amyotrophic lateral sclerosis (AL81 the nature of the first involvement of the neurons is not known. We examined the motor neuron0 of the triganinal nerve which usually show late involrcmrmt. Tbera *a8 nuole8r condexmation of ChrcmBtin due to increase in heterochrometin and lose of euchroartin, o-s that am consistent with a special degenerative process known as apoptosie. Only 40 pg RNA could be extracted from the8e neurons in contrast to 150 pg from controla. In situ hybridization of lllRNAto the midsize neurofilmmnt protein was evaluated by autor8diograpby Olig0IMCleOtida
with an antisense (54 mr), labeled
S-35 on the 3' end.
with
QUANTIFYIUG OF TAD-PROWS BY XB&HS OF AN ENZYME LINKBD ImVW0S0R~ltlW ASSAY
(ELISA)
K.Krohn, M.K.BriXkner and Th.Arendt Dept. Neurochem., Paul Flechsig Inst. Research, University Leipzig, Germany
Brain
The characteristic of cytoskeletal changes in Alzheimer Disease (AD) is attributed at least in part to changes in microtubule associated proteins (MAP's). For tau, a multible group of MAP's within a range of low molecular weight (45-70kDa). alterations in the state of phosphorylation might be an important step during pathogenesis. In addition, it has been shown that abnormal phosphorylated tauproteins are major constituents of paired helical filaments (PHF) implying that alterations in the amount of tau-proteins would provide a valuable marker for pathological changes. We purified tau-proteins from neuronal tissue of normal and AD brains to introduce an enzyme linked immunosorbens assay for tau proteins using monoclonal antibodies against a PHF preparation.
5
RXRRES8ION OF NQF AND BDNF IN AN AllHlAL YODlcL Or CSRONIC NXURONAL F-IN ~IZONIRTRBBRSAL M.K.Brikkner,
s.Pagliusi (*) and Th.Arendt Paul Flechsig Inst. Brain Dept. Neurochem., Research, University Leipzig, Germany and (*) Glaxo Inst. Molec. Biol. S.A., Geneva, Switzerland Neuronal injury and cell death in the basal nucleus both in Alzheimer's disease and after chronic intake of ethanol in human and rat appears to be associated with a plastic adaptive rasponse resulting in a remodelling of the dendritic organization. This process which is associated with both degeneration of dendrites and sprouting of new dendritic elements is probably mediated by trophic signals derived from afferent and efferent neuronal connections and from neuroglia. In the precompared the sent study, we have therefore pattern of neuronal degeneration and reorganization in the basal forebrain of rat brain induced by chronic treatment with ethanol with the pattern of the expression of nerve growth factor (NGF) and brain derived growth factor (BDNF) obtained in the basal nucleus and its target regions such as neocortex and hippocampus by means of mRNA northern blot technique and in situ hybridisation.
B2
6 MEASURING INOSITOL PHOSPHATES AND CDP-DIACYLGLYCEROL REVEALS DIFFERENT MECHANISMS FOR RECEPTOR ACTIVATION OF PHOSPHOLIPASE C E. Claro1r2, John N. Fai;', F. Picatiste' Departments of Biochemistry, University of Tennessee, Memphis, USA~, and Universidad Aut&oma de Barcelona, Spain2 Carbachol (CCH), noradrenaline (NA) and ionomycin (ION) Stimulated accumulation of inositol phosphates (InsP's) and CDP-diacylglycerol (CDP-DAG) were studied in rat corti_ cal slices. Relative to maximally effective concentrations of CCH (ImM), the InsP's responses due to NA (3OuN) and ION (3uM) were 231% and 3256, respectively, whereas the effi cacies of NA and ION towards the CDP-DAG response were 63% and 29% of that of CCH. Both InsP's and CDP-DAG are produced upon activation of phospholipase C (PLC), but the latter is formed after diacylglycerol in a rea'ction which is inhibited by calcium. Also, in contrast to CCH, NA is unable to stimulate PLC in cortical membranes in the presence of GTP(S) . These results suggest that, unlike muscarinic receptors, which are directly coupled to PLC through a G protein, alpha, adrenergic receptors stimulate PLC indirectly most probably by raising intracellular Ca2+ concentration.
8
7 ~OP~sIS
MOTOR
AI00 )(IcSSRNGER RNA IN TRIGE==NRWRONS INALS.
11.A.8~rtIMnn', S. Ton&c*' and D.Y. Sun'. ‘Dapaarmm+ of Pathology, University of Wisconrin,
Madison, WI, 53706 'RockefellerUniversity, New York, 10021, USA
N.Y.
In amyotrophic lateral sclerosis (AL81 the nature of the first involvement of the neurons is not known. We examined the motor neuron0 of the triganinal nerve which usually show late involrcmrmt. Tbera *a8 nuole8r condexmation of ChrcmBtin due to increase in heterochrometin and lose of euchroartin, o-s that am consistent with a special degenerative process known as apoptosie. Only 40 pg RNA could be extracted from the8e neurons in contrast to 150 pg from controla. In situ hybridization of lllRNAto the midsize neurofilmmnt protein was evaluated by autor8diograpby Olig0IMCleOtida
with an antisense (54 mr), labeled
S-35 on the 3' end.
with
QUANTIFYIUG OF TAD-PROWS BY XB&HS OF AN ENZYME LINKBD ImVW0S0R~ltlW ASSAY
(ELISA)
K.Krohn, M.K.BriXkner and Th.Arendt Dept. Neurochem., Paul Flechsig Inst. Research, University Leipzig, Germany
Brain
The characteristic of cytoskeletal changes in Alzheimer Disease (AD) is attributed at least in part to changes in microtubule associated proteins (MAP's). For tau, a multible group of MAP's within a range of low molecular weight (45-70kDa). alterations in the state of phosphorylation might be an important step during pathogenesis. In addition, it has been shown that abnormal phosphorylated tauproteins are major constituents of paired helical filaments (PHF) implying that alterations in the amount of tau-proteins would provide a valuable marker for pathological changes. We purified tau-proteins from neuronal tissue of normal and AD brains to introduce an enzyme linked immunosorbens assay for tau proteins using monoclonal antibodies against a PHF preparation.