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Metabolic profile of various neural cells: A basis for interpreting 1H NMR brain spectra in vivo
DS
9thMeeting oftheESN
17 pmsboLoQfcAcEMANCEOF THEPRO-ANDANTIOXlDATIVE REACTANTBiq/VfT.C
/ VlT.E IN BFtAlNCELLB
18
NBTABOLIC PROIILB OF VARIOUS CELLS: A BASIS FOR BIIW’Brn BRAIN
M. Uhr,F. Prel and H. Reiber NeumchemlfxhesLabor,UnlverskyG&Hng8n.Germ8ny The klnetlcs of ot-towpheml (vttamln E) and ascoMte (vlkmln C) oxkMon wlth braincell membraneprepaattaw (1.5 mg prctetn/mlas88y,eMogenousiron=lnrnoI and VR.EmO.1nmd, exogwm~ asubate=50-5000 nmol.at pH 7.2) w the fokwing m to be of physiologk&r&mnc8ln~:1.PnxxkMkefunction d Fe(H),2. prom&Mm fundbn of VttC by mcydlngFe(lll)to Fe@), 3. an@x&&~ functbn of Vlt.C at concentmtbns> 700 nmd/nli, 4. alukM&e fun&in of Vlt.E In the pmsence of Vlt.C,5. M.C rqctlng of Vlt_E.In addltbn the antioxkknt ftJMbtldVk.EkfolJndtobe competWvewlthVlt.C,butonly of endogenousIron abov8theequkrkntconcMmM and~tw8ntyfotdmore8ffectkthanVlt.C. Thelnvitro~ofheshlypreperedpigbraindigodond~ahovmdthatttw ~concentmtlonof Vlt.C > 1000 M/ml w88 amnt, but became prodecre8w of Vlt.C concenoxldsnt~lyJI~eubsequent tmncntoonmol/ml8twu8m8l8ndkrtemaim8mbr8neside. In contmt vital o&&mdnxytes incubatedw&h external 5&wnol/ml Wt. C mmalrwdst8bte agelnstIlpidpemxkktlon. TNs~cfcen~wIUbedkcussed~ rapecttothelnvlvoextr8&lular~ascorfHte B (> 200 nmol/ml) and the hlgh lntmceJlul8r This sensltlvebalance antMdantaso&8te B In gllnlcelfsmlghtbe disturbedIn MottxwParkinson. 19
V~AH)OXYTOUNMlUCEDELEVATCNSOF lw FREE 0+?+ lN SvMpATHEflC QAMLlA M. Verh8ge.W.H. &pen and P.N.E. de Graan Ruddf M8gnu8 lnet., Univ. Utrecht, The Netherknds The actlone of vlr(lopre8dn 8nd Oxytodnw8fe studled with dynamic diglt8l hnaglng of Intracellular free &?+ ([Qk) hi fwe-2 kided eymp8thetk g81@i8 and Mvldual cek from thee81gengtla. V-In (O.l-1pM) evokes a small [Call-elevation (20-30% overb888flev8f8)Inthelnt8ctg8nglbn.Dkeod8non ofthecekfmmtheg8n@onytefd8eever8lmorphologfo8Wyand lmm-ICatty dl8ttnguteh8ble oeflS kr arl(ue. V-n ewkes [C8~-~V8tkMl8 ln 8 v of th8 df88od8ted cell8 with ED50 between 10 8nd 100nM. The twpcnelv8 ceft8 are small splndle-shaped bl- or trlpolsr’ cells, immunoneg8threfor tfw8e f6rme of net&flament and do not respond to carb8chol or 8erotonln (up to lO@tM). The [C8h-ekV8tlon8 c8n 8till be evoked after buffering ICab to ~lctcmM, but not 8fter 1pM thapslgllrgfn, 8n IMbltor Of C8*+-tr8n8pMt In lntefll8l etwee. oXytIX&l 81#, evoke8 [C8Jt+fev8tion8 Inthe88me~ofcefk,butwith~lith~ hbhwED~o.Thedfucbdmtsqwmh are no8-8Mllve.The eifecte of v88opre88in c8n be bkcked with 8 SD8C& vj-type 88trgonla(O.l-1pM) butnotwith qedffc V2-type 8I'td oxytodnantagonists.
SPECTRA
t#BWRAL
IN VIVO.
J. Urenjak, S.R. Williams, D.G. Gadian, and M. Noble Department of Biophysics, The Royal College of Surgeons, London, U.K. Changes in 'H NMR spectra of the human brain are difficult to interpret because NMR does not discriminate the various neural cell populations. The purpose of this study was to examine the 'H NMR profile of the following cultured cells: cerebellar granule neurons, cortical type-l astrocytes, oligoastrocyte (O-2A) dendrocyte-type-2 progenitor cells, oligodendrocytes and meningeal cells. Amino acids and Ndata were acetyl-aspartate (NAA) verified by HPLC analysis. The data suggest that each individual cell type has a characteristic metabolic pattern that can be distinguished hy 'H NWR. A large amount of NAA was found in neurons, thus confirming its potential as a neuronal marker, but also in O-2A progenitor cells, an oligodendrocyte precursor cell. Analogies exist between the spectra of these cultured cells and those of corresponding brain tumours. 20 NRUR0CHRMICAL AND IMmNOcYmHRNICAL IDENTIFICATION OF GABA IN THE DORSAL ROOT GANGLIA (DRG) .
RAT
J. Weil-Pugazza *, A. Bertrand *, E. Phillipe* and G. Audet*. * INSERMU.161, Paris, France; c* Centre de Recherche en Neurobiologie, Quebec,Canada. Among the various amino acids detected in the DRG, gamma-aminobutyric acid (GABA)has been detected sensory neurons
in in
2 the
subpopulations chick DRG.
of
In the present study we report the neurochemical and inmunocytochemical detectionof GABA in the rat DRG. In the cervicalDRG the level of GABA, measuredby HPLC-ED,was 0.216 f 0.055 ppol/mg protein. This low level is corroborated by the iaxnunocytochuical identification of GABA in the DRG. Indeed only a very small numberof largeA neurons and occasional satellite cells were imnunostained.These observationssuggest that GABA may act as inhibitorytransmitter releasedby peripheralafferentfibers. It could also be possible that GABA act as neurotrophic factor with peripheral tissues. (Supportedby INSRRW,Franceand Freidreich AtaxiaAssociationof Canada)
9thMeeting oftheESN
17 pmsboLoQfcAcEMANCEOF THEPRO-ANDANTIOXlDATIVE REACTANTBiq/VfT.C
/ VlT.E IN BFtAlNCELLB
18
NBTABOLIC PROIILB OF VARIOUS CELLS: A BASIS FOR BIIW’Brn BRAIN
M. Uhr,F. Prel and H. Reiber NeumchemlfxhesLabor,UnlverskyG&Hng8n.Germ8ny The klnetlcs of ot-towpheml (vttamln E) and ascoMte (vlkmln C) oxkMon wlth braincell membraneprepaattaw (1.5 mg prctetn/mlas88y,eMogenousiron=lnrnoI and VR.EmO.1nmd, exogwm~ asubate=50-5000 nmol.at pH 7.2) w the fokwing m to be of physiologk&r&mnc8ln~:1.PnxxkMkefunction d Fe(H),2. prom&Mm fundbn of VttC by mcydlngFe(lll)to Fe@), 3. an@x&&~ functbn of Vlt.C at concentmtbns> 700 nmd/nli, 4. alukM&e fun&in of Vlt.E In the pmsence of Vlt.C,5. M.C rqctlng of Vlt_E.In addltbn the antioxkknt ftJMbtldVk.EkfolJndtobe competWvewlthVlt.C,butonly of endogenousIron abov8theequkrkntconcMmM and~tw8ntyfotdmore8ffectkthanVlt.C. Thelnvitro~ofheshlypreperedpigbraindigodond~ahovmdthatttw ~concentmtlonof Vlt.C > 1000 M/ml w88 amnt, but became prodecre8w of Vlt.C concenoxldsnt~lyJI~eubsequent tmncntoonmol/ml8twu8m8l8ndkrtemaim8mbr8neside. In contmt vital o&&mdnxytes incubatedw&h external 5&wnol/ml Wt. C mmalrwdst8bte agelnstIlpidpemxkktlon. TNs~cfcen~wIUbedkcussed~ rapecttothelnvlvoextr8&lular~ascorfHte B (> 200 nmol/ml) and the hlgh lntmceJlul8r This sensltlvebalance antMdantaso&8te B In gllnlcelfsmlghtbe disturbedIn MottxwParkinson. 19
V~AH)OXYTOUNMlUCEDELEVATCNSOF lw FREE 0+?+ lN SvMpATHEflC QAMLlA M. Verh8ge.W.H. &pen and P.N.E. de Graan Ruddf M8gnu8 lnet., Univ. Utrecht, The Netherknds The actlone of vlr(lopre8dn 8nd Oxytodnw8fe studled with dynamic diglt8l hnaglng of Intracellular free &?+ ([Qk) hi fwe-2 kided eymp8thetk g81@i8 and Mvldual cek from thee81gengtla. V-In (O.l-1pM) evokes a small [Call-elevation (20-30% overb888flev8f8)Inthelnt8ctg8nglbn.Dkeod8non ofthecekfmmtheg8n@onytefd8eever8lmorphologfo8Wyand lmm-ICatty dl8ttnguteh8ble oeflS kr arl(ue. V-n ewkes [C8~-~V8tkMl8 ln 8 v of th8 df88od8ted cell8 with ED50 between 10 8nd 100nM. The twpcnelv8 ceft8 are small splndle-shaped bl- or trlpolsr’ cells, immunoneg8threfor tfw8e f6rme of net&flament and do not respond to carb8chol or 8erotonln (up to lO@tM). The [C8h-ekV8tlon8 c8n 8till be evoked after buffering ICab to ~lctcmM, but not 8fter 1pM thapslgllrgfn, 8n IMbltor Of C8*+-tr8n8pMt In lntefll8l etwee. oXytIX&l 81#, evoke8 [C8Jt+fev8tion8 Inthe88me~ofcefk,butwith~lith~ hbhwED~o.Thedfucbdmtsqwmh are no8-8Mllve.The eifecte of v88opre88in c8n be bkcked with 8 SD8C& vj-type 88trgonla(O.l-1pM) butnotwith qedffc V2-type 8I'td oxytodnantagonists.
SPECTRA
t#BWRAL
IN VIVO.
J. Urenjak, S.R. Williams, D.G. Gadian, and M. Noble Department of Biophysics, The Royal College of Surgeons, London, U.K. Changes in 'H NMR spectra of the human brain are difficult to interpret because NMR does not discriminate the various neural cell populations. The purpose of this study was to examine the 'H NMR profile of the following cultured cells: cerebellar granule neurons, cortical type-l astrocytes, oligoastrocyte (O-2A) dendrocyte-type-2 progenitor cells, oligodendrocytes and meningeal cells. Amino acids and Ndata were acetyl-aspartate (NAA) verified by HPLC analysis. The data suggest that each individual cell type has a characteristic metabolic pattern that can be distinguished hy 'H NWR. A large amount of NAA was found in neurons, thus confirming its potential as a neuronal marker, but also in O-2A progenitor cells, an oligodendrocyte precursor cell. Analogies exist between the spectra of these cultured cells and those of corresponding brain tumours. 20 NRUR0CHRMICAL AND IMmNOcYmHRNICAL IDENTIFICATION OF GABA IN THE DORSAL ROOT GANGLIA (DRG) .
RAT
J. Weil-Pugazza *, A. Bertrand *, E. Phillipe* and G. Audet*. * INSERMU.161, Paris, France; c* Centre de Recherche en Neurobiologie, Quebec,Canada. Among the various amino acids detected in the DRG, gamma-aminobutyric acid (GABA)has been detected sensory neurons
in in
2 the
subpopulations chick DRG.
of
In the present study we report the neurochemical and inmunocytochemical detectionof GABA in the rat DRG. In the cervicalDRG the level of GABA, measuredby HPLC-ED,was 0.216 f 0.055 ppol/mg protein. This low level is corroborated by the iaxnunocytochuical identification of GABA in the DRG. Indeed only a very small numberof largeA neurons and occasional satellite cells were imnunostained.These observationssuggest that GABA may act as inhibitorytransmitter releasedby peripheralafferentfibers. It could also be possible that GABA act as neurotrophic factor with peripheral tissues. (Supportedby INSRRW,Franceand Freidreich AtaxiaAssociationof Canada)