(Ch inese Journa l of Neuroana tom y)

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1 Sep (3) (Ch inese Journa l of Neuroana tom y) (, ),, CONSTRUCTING A PARTIALLY L ESIONED PARK INSON D ISEASE MOD EL OF RAT BY TRANSECTING THE M ED IAL FOREBRA IN BUNDL E W ITH A W IRE KN IFE L iang X ibin, L uo Y ong, L u J un, L iu X iany u, Z hang W angm ing, L i F eng qiao, W ang X iaom in, H an J isheng (N euro science Research Institute, Peking U niversity, Beijing ) Abstract T he m edial fo rebrain bundle of rats w as transected w ith a w ire knife to set up a model of ch ronic degeneration of dopam inergic neurons. The abno rm al ro tation of rats, the extent of the substantia nigra neurons dam age and the amount of dopam ine in striatum w ere investigated by behavio ral test, ho rseradish peroxidase retrograde tracing, tyro sine hydroxylase im 2 m unohistochem istry and high perfo rm ance liquid chrom atography at different tim e after operation. The results indicated that the partial lesioned model of Parkinson s disease can be successfully established by transecting the m edial fo rebrain bundle of rats w ith a w ire knife. (F igures 5, 6 on p late 35) Key words w ire knife, m edial fo rebrain bundle, partial lesion, parkinson disease model, rat (PD ),, 62OH 2 (DA ), DA DA (m edial fo reb rain bundle,, M FB ) PD DA,, PD DA DA, PD PD, (N o. G ), (N o ) (N o )

2 M FB DA PD 2, 3 d) DA, 4. 2 (Am phetam ine, Am p ) PD 24, 48 h, PD, PD 4. 3 HR P M FB HR P, 1. W istar ( ), g, %,, 2., ( ),, 50 Λm, TM B (Sigm a), HR P ( ), 4. 4 ( TM B 2HC l, Sigm a ), : DA, DO PA C, HVA, 52H IAA (Sigm a) H PL C 3 :A 2 (0. 15 m o lgl,, 15 Λm, PBS ( ph 4. 3) : g, m o lgl, ph 7. 4), 5 m in 0. 3% H 2O 2, g, dh 2O 500 m l; B 37, 15 m in PBS, 5 m in 3 10% ED TA 2 N a (0. 1 m o lgl ) : ED TA N a g, dh 2O 100 m l; C ), 4, 48 h PBS, 5 m in 3 (120 mm o lgl ) : g, dh 2O, m l 3. 5 : , : 1. 2 mm, 2. 5 mm, 5. 0 mm 30% HR P 4 5 Λl, m in, 15 m in, 3 d,, 4%, 2 h, 20% M FB 4,, 37, 15 m in, ( ,, 37, 30 m in PBS, 5 m in 3 HR P, 37, 30 m in PBS, 5 m in % DAB - 3% H 2O 2 2 Scou tern ( Kopf m in M ayo r 2 m in, ) ( ),, ( ) ECD 26A 6 TH, 4. TH 4. 1, [ 1, 2 ] Scou tern 4. 5 M FB 6% (350 m ggkg, i. p. ) U ngerstedt [ 3 ], Am p (5 m ggkg),, 3% H 2O 2, 45 m in, 2 5 m in,, : 3. 8 mm 2. 4 ( ) (,, ) 45 m in ( g Scou tern 8. 0 mm, m in), mm, mm,, 3,, (10 m gg100 g, h,

3 , cro soft Excel 97,, (x θ S xθ) Pow erpo in t - 80 P rism ( H PL C2ECD (ANOVA s) N ewm an Keu ls po st2hoc ) - 80, 1. 5 m l Eppendo rf 200 Λl A, 10 s (1H z, 1 s, : 201 P < s, 4 5 ), 1 h, 4 1. HR P rgm in 20 m in 160 Λl, HR P ( DA B, ), DA 1 h, rgm in 20 m in, M FB DA DA F ig. 5, [4 ] 1. 2 m lgm in,l 2ECD 26A, M FB V, 1 na gmv, 50 mv, 10 2, 2. F ig. 6 M FB 4, DA,, 6 TH,,, ( TH, ),,M FB 4 60% DA 5. (F ig. 1) M FB M i2 DA F ig. 1 T he percentage of TH po sitive neurons in the SN c and V TA on the lesio ined side versus unlesioned side. 3. ( ) Am p gm in,, M FB 2,, DA gm in M FB 3 gm in,

202 17 3 2001 9 (F ig. 2) 2, 1 (1g10) ; 6 1 (1g8) ; 8 2 M FB DA ( 2g11) ; 10 4 (4g10) 2 g 2 gm in, m in 2 gm in 84. 3% DA, 2 F ig. 2 Ro tation sco res after M FB transection 3, P < 0. 05; 3 3, P < 0.

4 (F ig. 2) 2, 1 (1g10) ; 6 1 (1g8) ; 8 2 M FB DA ( 2g11) ; 10 4 (4g10) 2 g 2 gm in, m in 2 gm in 84. 3% DA, 2 F ig. 2 Ro tation sco res after M FB transection 3, P < 0. 05; 3 3, P < in comparison w ith the naive. 4. H PL C2ECD DA, 2, F ig. 3 H PL C2ECD, DA, DA, 40% 60% (F ig. 4), 30, M FB, m in, 0. 5 DA, 16 ng DA, 8 ng 16 ng, H PL C PD DA F ig. 3 Ch rom atogram of monoam ines and their m etabo lism s.

5 : 203 F ig. 4 Dopam ine contents in rat striatum after M FB transection 3, P < 0. 05; 3 3, P < 0. 01; 3 3 3, P < in comparison w ith the unlesioned side. PD DA, 30% 40%,, DA DA PD DA 90%, [ 5 ] DA DA 1988 Jan son 2, M FB [ 12 ] (partial hem itran2, section) DA DA, 15 d 52% DA M FB,, PD, DA (A pom o rph ine, A po ) DA Am p [ 3, 6, 7 ], L apchak [ 14 ] : 1. DA,,, DA, DA DA,, DA, M FB 18 A po d 19 d 44% 50% DA 2. DA Am p DA,, DA 2 [ 7210, ] PD DA,,, DA, 62OHDA PD, M FB PD : PD (1) 62OHDA 10 m in DA, 3 d 95% DA [ 11 (2) ] 62OH 2 DA (3),, Knu sel [ 13 ] M FB DA 40% 50% ( ) 5, 6 35

6 [ 1 ] T seng JL, Baetge EE, Zurn AD, et al. GDN F reduces drug2in2 duced ro tational behavio r after m edial fo rebrain bundle tran2 section by a m echanism no t invo lving striatal dopam ine. J N euro sci, 1997, 17: [ 2 ] B recknell JE, D unnett SB, Faw cett JW. A quantitative study of cell death in the substantia nigra fo llow ing a m echanical le2 sion of the m edial fo rebrain bundle. N euro science, 1995, 64: [ 3 ] U ngerstedt U, A rbuthnett GW. Q uantitative reco rding of ro ta2 tional behavio r in rats after 62hydroxydopam ine lesions of the nigro striatal dopam ine system. B rain Res, 1970, 24: [ 4 ] Felice LJ, Felice JD, Kissinger PT. D eterm ination of cate2 cho lam ines in rat brain parts by reverse2phase ion2pair liquid ch rom atography, J N eurochem, 1978, 31: [ 5 ] Fahn S, L ibsch L R, Catler RW. M onoam ines in the hum an neo striatum: topographic distribution in no rm al and in Parkinson s disease and their ro le in akinesis, rigidity, cho lera and tremo r. J N euro l Sci, 1971, 14: [ 6 ] U ngerstedt U. Striatal dopam ine release after amphetam ine o r nerve degeneration revealed by ro tational behavio r. A cta phys2 io l Scand Supp l, 1971, 367: [ 7 ] U ngerstedt U. Po stsynap tic supersensitivity after 62hydroxy2 dopam ine induced degeneration of the nigro striatal dopam ine system. A cta Physio l Scand, 1971, 367 (supp l) : [ 8 ] C reese I, Burt DR, Snyder SH. Dopam ine recep to r binding en2 hancem ent accompanies lesion2induced behavio ral supersensi2 tivity. Science, 1977, 197: [ 9 ] Schultz W, U ngerstedt U. Sriatal cell supersensitivity to apo2 mo rphine in dopam ine2lesioned rats co rrelated to behavio ṙ N europharm aco l, 1978, 17: [ 10 ] H udson JL, V an Ho rne CG, Strom berg I, et al. Co rrelation of apomo rphine2 and amphetam ine2induced turning w ith nigro s2 triatal dopam ine content in unilateral 62hydroxydopam ine le2 sioned rats. B rain Res, 1993, 626: [ 11 ] Schm idt RH, B jo β rk lund A, Stenevi U, et al. Intracerebral grafting of neuronal cell suspension. III, A ctivity of intrastrial nigral suspension imp lants assessed by m easurem ents of dopam ine synthesis and m etabo lism, A cta Physio l Scand, 1983, 522 (supp l) : [ 12 ] Janson AM, Fuxe K, A gnatil F, et al. Ch ronic nico tine treat2 m ent counteracts the disappearance of tyro sine hydroxylase2 imm uno reactive nerve cell bodies, dendrites and term inals in the m eso striatal dopam ine system of the m ale rat after partial hem itransection. B rain Res, 1988, 455: [ 13 ] Knusel B, Beck KD, W inslow JW, et al. B rain2derived neu2 ro troph ic facto r adm inistration p ro tects basal fo rebrain cho lin2 ergic but no t nigral dopam inergic neurons from degenerative changes after axo tom y in the adult rat brain. J N euro sci, 1992, 12: [ 14 ] L apchak PA, Beck KD, A raujo DM, et al. Ch ronic intranigral adm inistration of brain2derived neuro trop ic facto r p roduces striatal dopam inergic hypofunction in unlesioned adult rats and fails to atttenuate the decline of striatal dopam inergic function fo llow ing m edial fo rebrain bundle transection. N euro science, 1993, 53: Explana tion of F igures F ig. 5 HRP retrograde labeling of the dopam inergic neurons in no rm al(upper) and M FB L esioned (m iddle) rats. A, no rm al rat w ith no2hrp injected B, HRP injected in no rm al rat C, U nlesioned side of the M FB transected rat w ith no2 HRP injected D, HRP injected into the lesioned side of them FB lesioned rat E, m agnification of B F, m agni2 fication of D Scale bar= 100 Λm in A, B, C, D and 20 Λm in E, F F ig. 6 TH imm unoh istochem ical staining show ing the dopam inergic neurons in the V TA (ventral tegm ental area) and SN c (sub2 stantia nigra compacta). A, naive group B, M FB lesioned group

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