laminae III-IV par l’intermédiaire d’un réseau glial. Des résultats préliminaires semblent indiquer l’implication d’une communication au sein d’un réseau glial car la 08,793-V->-3/'( K<2( 7>-K</( >/5( 0838<V( .-3012-33/>5'( 7>-K</( 9J8>/;/31( >_/66/1( 4/( >8( 0>-32423/( 5<,( >8( 1,835;2552-3( 5D38:12K</( 23E2721,20/( 4835( >/5( >8;238/( XXX-IV. Nous sommes en train de mettre en place un poste d’imagerie du calcium sur tranches de moelle épinière qui devrait nous permettre de déterminer si des vagues de calcium intracellulaire se propagent des laminae profondes vers les laminae III-IV au sein du réseau glial suite à l’application locale d’agonistes de la NA dans les laminae profondes. Le cas échéant, nous vérifierons si cette propagation dans le réseau glial est bloquée par la carbénoxolone. Dans ces expériences, la propagation d’information dans les réseaux 4/( 3/<,-3/5( 5/,8( 7>-K<9/( /3( 8.-<1831( 4/( >8( \\|( 8<( ;2lieu extracellulaire afin de 3_-75/,=/,( K</( >/5( 0-;;<320812-35( 4835( >/( ,95/8<( J>28>$( `/5( 851,-0D1/5( :-<,,-31( n1,/( visualisés après une incubation des tranches avec de la sulforhodamine Ok8621j(/1(8>$'( "GGHN, <3(;8,K</<,(6><-,/50/31(K<2(5/(62V/(5<,(>/5(851,-0D1/5(=2=8315$(M>1/,3812=/;/31( on pourrait étudier les changements de calcium intracellulaire sélectivement dans les 851,-0D1/5(par l’intermédiaire d’une sonde calcium encodée géné12K</;/31(Ol08;:#N(/1( exprimée sous le promoteur de la GFAP OuE2J/1-;2(/1(8>$'("G!#N.
("Identification des messagers impliqués dans la
communication glie-neurone
Nos résultats suggèrent que le glutamate et/ou l’ATP pourraient servir de J>2-1,835;/11/<,5(4835(3-1,/(5D51I;/(:-<,(855<,/,(>8(0-;;<320812-3(J>2/-neurone suite à la stimulation par les agonistes de la NA. L’utilisation de cocultures de neurones/glie 5:238>/5(O;25/5(8<(:-231(/1(<12>259/5(/3(,-<123/(4835(3-1,/(>87-,81-2,/N(3-<5(:/,;/11,8( d’étudier dans des conditions d’approche pharmacologique idéales la libération d’un gliotransmetteur par la glie et son action sur les neurones =-25235(O9>9;/315(491/01/<,5(
!G* du gliotransmetteur libéré)$(S-<5(:,9=-D-35(9J8>/;/31(>_<12>25812-3(4/(0/>><>/5(oBk("L#( 1,8356/019/5( 8=/0( 4/5( ,90/:1/<,5( +"|"( 4/( >_M\+( qui nous servirons d’éléments de détection de l’ATP O53266/,(0/>>N(4/(>_M\+$(
)"Stimulation optogénétique des fibres
noradrénergiques
Une infection du LC avec un virus permettant l’expression de la 0E833/>,E-4-:523/( "( 5-<5( >/( 0-31,r>/( 4<( :,-;-1/<,( 4/( >8( 4-:8;23/( &-ED4,-VD>85/( permettrait une activation optique des fibres noradrénergiques sur des préparations in
vitro de moelle épinière de rat. De telles expériences devraient idéalement reproduire
>/5(,95<>1815(-71/3<5(8=/0(>_8::>20812-3(4/(3-,1,2:1D>23/(O7>-K<831(4/(>8(,/08:1<,/(4/(>8( NA) ou l’application exogène d’agonistes de la NA. L’approche optogénétique nous permettrait également de mieux caractériser la transmission noradrénergique en enregistrant et en caractérisant pharmacologiquement les variations de potentiel de ;/;7,83/( O++uB'( ++SI) induits par la stimulation optogénétique des fibres noradrénergiques dans des neurones ou des cellules gliales de la CDME enregistrées en configuration cellule entière de la technique du patch-clamp.
!GC
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