1Bogomoletz Institute of Physiology, Kiev, Ukrainenana@biph.kiev.uaDr. NANA VOITENKODr. NANA VOITENKOAACIMPAACIMP KIEV - 2011KIEV - 2011Molecular Mechanisms of Pain part II2Part B!"pinal #$P#%s!me&iate& synapti' transmission &uring inflammation. !$ole'ular me'hanism of (lu%2 internalization from synapses.Part )!Involvement of spinal e*trasynapti' #$P#%s in the maintaining of persistent pain.!+raffi'king of )a2,!permeable #$P#%s &uring inflammatory pain. Part - ! Protein kinase )a as a mole'ular target for perspe'tive treatment of persistent pain. ! #ntisense oligonu'leoti&es . a ne/ strategy of pain treatmentPart # !)lassifi'ation of re'eptors to glutamate.!"tru'ture an& mole'ular organization of #$P# re'eptors.0GLUTAMATE RECEPTORSIonotropic GluRs Metabotropic GluRsNMDANR1NR3ANR2A-DAMPAGluR 1-4KainateGluR 5,6, KA 1,2Group I!Glu1!Glu5Group II!Glu2!Glu3Group III!Glu4!Glu6!Glu"!Glu

-rug 1e/s Perspe't 2220, 13456 710Meia!in" effec!s inclue#! $embrane &epolarization! )a2, an& 1a, influ*! K, efflu*! 8ree ra&i'al formationMoula!or$ effec!s on#! i(lu%! K, an& )a2, 'hannels ! 1eurotransmitter release9#$P# re'eptorsisasubtypeofionotropi'transmembranere'eptorfor glutamate that me&iates fast synapti' transmission in the 'entralnervoussystem.Itsnameis&erive&fromits abilitytobea'tivate&bytheartifi'ialglutamateanalog #$P#.+here'eptor/as&is'overe&by+age:onore an&'olleaguesatthe"'hoolofPharma'yin )openhagen,an&publishe&in1;52intheJournalof Neurochemistry. #$P#%s are foun& in many parts of the brainan&arethemost'ommonlyfoun&re'eptorinthe nervous system. #$P#%saretraffi'ke&fromthe&en&riteintothe synapsean&in'orporate&throughsomeseriesof signaling'as'a&esin'reasingeffi'a'yofsynapti' transmission. 7$ole'ular 2 mG an& 92 mG hol&ing potentials.##$2#$4#$6#$"11$21$4# 2 4 6 " 1# 12 14 16%aline &'AP()* 433Nor!ali+e,-P%&a!plitu,es)i!e .!in/##$2#$4#$6#$"11$21$4# 2 4 6 " 1# 12 14 16%aline &'AP()* 433Nor!ali+e,-P%&a!plitu,es)i!e .!in/-1$#-#$"-#$6-#$4-#$2#$##$2#$4-"# -6# -4# -2# # 2# 4#%aline&'ANor!ali+e,currentMe!brane potential .!0/PPG1KI 52466 %1M 2#"11# !s5# pA%aline &'A1# !s5# pA# B)#$P#%!me&iate& Hvoke& HP")s at "ynapses bet/een Primary #fferents an& "( 1eurons)opyright L222; "o'iety for 1euros'ien'ePark, M.!"., Goitenko, 1. et al. M. 1euros'i. 222;N2;0223!1;1>1$-# %e'eptors an& PK) 'ouple to #$P#% 'omple* PI)K1 'oimmunopre'ipitates /ith (lu%2 an& PK) (A), /hile (lu%2 ! /ith PI)K1 an& PK) (). P"-!;7 'oimmunopre'ipitates /ith 1%2B an& stargazin (C), /hile stargazin ! /ith P"-!;7 an& (lu%2 (D).E& 1%1 47 nm gol&, arro/hea&s6 an& (lu%2 417 nm gol&6 'olo'alize at -: synapses. "'ale bar, 122 nm.)opyright L222; "o'iety for 1euros'ien'ePark, M.!"., Goitenko, 1. et al. M. 1euros'i. 222;N2;0223!1;15Propose& $e'hanism for (lu%2 Internalization from "ynapses1;)on'lusion Persistent peripheral inflammation in&u'es (lu%2 internalization from synapses via 1$-# %e'eptor!triggere& PK) a'tivation in -: neurons22Part ) Involvement of spinal e*trasynapti' #$P#%s in the maintaining of persistent pain.+raffi'king of )a2,!permeable #$P#%s &uring inflammatory pain. 21)ombine& Hle'trophysiology an& )al'ium Imaging in Substantia Gelatinosa 1euron+ransmitte& light image of Substantia Gelatinosa in transverse -: sli'e 4left6N "(neuron loa&e& /ith 222 m$ of fura!2 4right6. 22#$P#%s!me&iate& )urrent an& Q)a2,Ri +ransients in "oma = -en&rites of "( 1eurons %G neurons 2ere i,enti3ie, accor,in4 to t(eir pattern o3 action potential 3irin4A, %i!ultaneous recor,in4 o3 AMPA-in,uce, current .lo2er ro2/ an, 5&a267i transients .upper ro2/ in so!a an, ,en,rites o3 %G neurons$ B-C, AMPARs anta4onist, N89* an, G1KI, abolis(e, current an, 5&a267i transients$D, %tatistics o3 current a!plitu,es .le3t/ an, 5&a267i transients .ri4(t/ in ,i33erent 4roups o3 %G neurons$Goitenko group, unpublishe& &ataKopa'h et. al. Pain, 2211. 20Inflammation Potentiates #$P#%s!me&iate& )urrent an& Q)a2,Ri +ransients in S+oni'T but not in S+ransientT "( 1euronsA& #$P#!in&u'e& 'urrent 4lo/er ro/6 an& Q)a2,Ri transients 4upper ro/6 in Stoni'T neurons 29h after saline or )8#. '-C& # s'atter &ot plot of 'urrents in "( neurons.D& "tatisti's of 'urrent amplitu&es 4left6 an& Q)a2,Ri transients 4right6 in "( neurons. E& # relationship of the Q)a2,Ri transient amplitu&es an& a normalize& value of integrate& 'urrent in the timeframe of #$P# appli'ation.-HKopa'h et. al. Pain, 2211. 29Inflammation!in&u'e& In'rease of the 1umber of H*trasynapti' )a2,!permeable #$P#%s A. :e3t, AMPA-in,uce, currents a3ter 5 !in pre-application o3 I-M at 24 ( post-&'A or saline$ Ri4(t, I-M 2as applie, ,urin4 stea,;-state current letonic? neurons at 1 , post-saline an, &'A$ Note, &'A-in,uce, in2ar, recti3ication 2as co!pletel; retonic? neurons 3ro! 1 , saline- an, &'A-treate, rats$ Kopa'h et. al. Pain, 2211. 27+he #ltere& Cevel of (lu%1 an& (lu%2 in the Plasma $embrane 8ra'tion = )ytosol after )8#Park et al., $ol. Pain, 2225 Kopa'h et. al. Pain, 2211. 23Peripheral Inflammation In&u'es (lu#1 $embrane Insertion at H*trasynapti' "ites of "( 1euronsA$ %ur3ace e=pression o3 GluA1 in DA neuron 1 , post-&'A or saline$ )op, Representati