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Regulation of phospholipase D activity by muscarinic receptors
B.5
9thMeeting
17
of the ESN
18
REQULATIOB
CHEXCAL COMPONkNTS OF CBEBRAL WHITE MATTER IN ALZHEIMER DISEASE M. Wender, J. Szc4rech,J. Dorseerska Department of Neurolo Medioal School, PoaX'Poland The studies were performed on autopey material of patients who died at the age above 70 years. The material was divided into the subgroup of brains with vascular changes only and the subgroup with senile atrophy of Alzheimer type /SDAT/. It was found the general decrease in myelin yield ae well as some minor ohanges in myelin lipid pattern.Changee in myelin proteins included a mild decreaee in Wolfgram protein con tent with reciprooal increase in large basic protein oontent.The degeneration of reasela is the decissive factor in the pathomechanism of myelin leaion.Using electron paramagnetic resonance analysis,was fo d di!Yand mi shed concentration of C!u+ "f ions and free radicals,concomiFe+ tant with increased concentration of more marked in multiions aggregates, cases of SDAT, reeulting from some slow down of metabolic processes. 19 PARTIAL PURIFICATION OF THE HUMAN TRANSPORTER AND PLATELET 5-HT CROSSREACTIVITY WITH ANTIBODIES TO THE PAT BRAIN GABA TRANSPORTER. H.M, * Mabjeesh, Worrall, D.M., Tarrant, N.J., Williams, D.C. and *Kanner, B.I. Department of Biochemistry, Trinity College, *Department Of Dublin 2, Ireland, and Biochemistry, Hadaszah Medical School, The Hebrew University, Jerusalem, Israel. The sodium dependent 5-HT transporter was partially purified from human platelets using Mono-Q anion exchange chromatography followed by affinity chromatography on aminomethyl-citalopram agarose. The protein fractions were subjected to SDS-PAGE and immonoblotting using an affinity purified polyclonal antibody to the rat brain GABA transporter. Crozzreactivity was found in all fractions with a protein of molecular weight 72 kD. This finding suggests that the S-HT transporter is substantially homologous to the GABA transporter and perhaps also to the similar noradrenaline transporter. Such homology has subsequently been confirmed with the elucidation of the cDN.4 sequence of the cloned 5-HT transporter from rat brain and rat basophilic leukemia cells.
ACTIVITY
BY
OF PBOSPBOLIP&SB l4USCARIlUC R%!BPTORS.
D
L. Gustavsson, M. Boyano-Adanaz and C. Lundqvist. Department of Psychiatry andNeurochemistry, University of Lund, Lund, Sweden. During recent years phospholipaae D has emerged as an important component of signal transduction cascades. The enzyme catalyses formation of phoophatidic acid and when ethanol is present production of phosphatidylethanol. Receptor-mediated phospholipase D activity was studied in the human neuroblastoma cell line SH-SY5Y after prelabelling with radioactive fatty acids. Phoephatidic acid, phosphatidylethanol and 1,2-diacylglycerol production was analysed. Carbachol induced a rapid and transient increase in phosphatidylethanol as well as phosphatidic acid via stimulation of muacarinic receptors. The products formed addition reached a maximum one minute after of the agonist. The increase in phosphatidic acid was substantially larger than the increase in phosphatidylethanol formation. Furthermore, the phosphatidic acid increase was not significantly inhibited by ethanol. This indicates involvement of other mechanisms in addition to phoapholipase D stimulation for the accumulation of phosphatidic acid after carbachol stimulation of SH-SYSY cells. 20
diUC&LLCAL CHAi&CTkRISTICtiOh ifLUTkLiICACID SYY'J!E& k'NZYLlES IN SYMAPTOSOL&j 02,VISUAL AlXLYYLR DURING DOG XAhLY POSTNATAL ONTOGLjlESfiS. T.FI;I.AGAXV A.l.Baraev Institute of Physiology kcad.of Sciences,Azerbaijan,Baku The first days after puppiers were born the glutamatdecarboq~lase, phosphate activated glutaminase,glutamine synthetase, GABA-transaminase, I-glutamyltransferase, alanine-and aspartate aminotransferase, with takes part in the metabolism of neuromediators-glutamate and GABA, were observed mainly in front doublement and external elbow body in comparasion with visual cortex (field Ii'). In the 12-16 postnatal days of enzymes is on maximum in synaptosomes of visual cortex, but till the 3 month of age activity is growing in sinaptosomes of all structures With is stipulated by formation of glutamate and GABA-ergic synaps in visual analyser of dogs brain. In the period from 3 to 6 month of age the activity of above mentioned enzymes in synaptosomes of structures of dogs brain in unchsnged or reducing.
9thMeeting
17
of the ESN
18
REQULATIOB
CHEXCAL COMPONkNTS OF CBEBRAL WHITE MATTER IN ALZHEIMER DISEASE M. Wender, J. Szc4rech,J. Dorseerska Department of Neurolo Medioal School, PoaX'Poland The studies were performed on autopey material of patients who died at the age above 70 years. The material was divided into the subgroup of brains with vascular changes only and the subgroup with senile atrophy of Alzheimer type /SDAT/. It was found the general decrease in myelin yield ae well as some minor ohanges in myelin lipid pattern.Changee in myelin proteins included a mild decreaee in Wolfgram protein con tent with reciprooal increase in large basic protein oontent.The degeneration of reasela is the decissive factor in the pathomechanism of myelin leaion.Using electron paramagnetic resonance analysis,was fo d di!Yand mi shed concentration of C!u+ "f ions and free radicals,concomiFe+ tant with increased concentration of more marked in multiions aggregates, cases of SDAT, reeulting from some slow down of metabolic processes. 19 PARTIAL PURIFICATION OF THE HUMAN TRANSPORTER AND PLATELET 5-HT CROSSREACTIVITY WITH ANTIBODIES TO THE PAT BRAIN GABA TRANSPORTER. H.M, * Mabjeesh, Worrall, D.M., Tarrant, N.J., Williams, D.C. and *Kanner, B.I. Department of Biochemistry, Trinity College, *Department Of Dublin 2, Ireland, and Biochemistry, Hadaszah Medical School, The Hebrew University, Jerusalem, Israel. The sodium dependent 5-HT transporter was partially purified from human platelets using Mono-Q anion exchange chromatography followed by affinity chromatography on aminomethyl-citalopram agarose. The protein fractions were subjected to SDS-PAGE and immonoblotting using an affinity purified polyclonal antibody to the rat brain GABA transporter. Crozzreactivity was found in all fractions with a protein of molecular weight 72 kD. This finding suggests that the S-HT transporter is substantially homologous to the GABA transporter and perhaps also to the similar noradrenaline transporter. Such homology has subsequently been confirmed with the elucidation of the cDN.4 sequence of the cloned 5-HT transporter from rat brain and rat basophilic leukemia cells.
ACTIVITY
BY
OF PBOSPBOLIP&SB l4USCARIlUC R%!BPTORS.
D
L. Gustavsson, M. Boyano-Adanaz and C. Lundqvist. Department of Psychiatry andNeurochemistry, University of Lund, Lund, Sweden. During recent years phospholipaae D has emerged as an important component of signal transduction cascades. The enzyme catalyses formation of phoophatidic acid and when ethanol is present production of phosphatidylethanol. Receptor-mediated phospholipase D activity was studied in the human neuroblastoma cell line SH-SY5Y after prelabelling with radioactive fatty acids. Phoephatidic acid, phosphatidylethanol and 1,2-diacylglycerol production was analysed. Carbachol induced a rapid and transient increase in phosphatidylethanol as well as phosphatidic acid via stimulation of muacarinic receptors. The products formed addition reached a maximum one minute after of the agonist. The increase in phosphatidic acid was substantially larger than the increase in phosphatidylethanol formation. Furthermore, the phosphatidic acid increase was not significantly inhibited by ethanol. This indicates involvement of other mechanisms in addition to phoapholipase D stimulation for the accumulation of phosphatidic acid after carbachol stimulation of SH-SYSY cells. 20
diUC&LLCAL CHAi&CTkRISTICtiOh ifLUTkLiICACID SYY'J!E& k'NZYLlES IN SYMAPTOSOL&j 02,VISUAL AlXLYYLR DURING DOG XAhLY POSTNATAL ONTOGLjlESfiS. T.FI;I.AGAXV A.l.Baraev Institute of Physiology kcad.of Sciences,Azerbaijan,Baku The first days after puppiers were born the glutamatdecarboq~lase, phosphate activated glutaminase,glutamine synthetase, GABA-transaminase, I-glutamyltransferase, alanine-and aspartate aminotransferase, with takes part in the metabolism of neuromediators-glutamate and GABA, were observed mainly in front doublement and external elbow body in comparasion with visual cortex (field Ii'). In the 12-16 postnatal days of enzymes is on maximum in synaptosomes of visual cortex, but till the 3 month of age activity is growing in sinaptosomes of all structures With is stipulated by formation of glutamate and GABA-ergic synaps in visual analyser of dogs brain. In the period from 3 to 6 month of age the activity of above mentioned enzymes in synaptosomes of structures of dogs brain in unchsnged or reducing.