大鼠脊髓后角内神经降压素对一级传入C纤维突触前抑制效应的形态学证据_英文_

大鼠脊髓后角内神经降压素对一级传入C纤维突触前抑制效应的形态学证据_英文_ M O RPHOLO G ICAL EV ID ENCES FO R PRESY NA PT IC INH IB ITO RY EFFECT O F NEURO TENS IN O N PR IM A RY A FFERENT C F IBER IN THE SP INAL DO RSAL HO RN O F THE RA T H e L i, Y ino n Zh an g , M in h a i Zh an g , Sh im in g Y an g, H o n g lian L i (, ,D ep a rtm en t of H is tology a n d Em bry ology T on g j i M ed ica l U n iv e rs ity ), 430030, . . W u h a n PRof C h in a () T h e a im of th e p resen t s tu dy is to rev ea l w h e th e r n eu roten s in N T in th e sp in a l d orsa l h orn of th e . , ra t m ig h t p resy n ap t ica l ly m od u la te th e p r im a ry af f e ren t C f ibe rsW ith f lu orescen ce m ic roscop eit w a s ob2 , 2 , 2? I se rv ed th a tin th e lam in a e of th e sp in a l cord th e d is t r ibu t ion of n eu roten s in l ik e im m u n orea c t iv ity () ( 24 2 N T L I w a s p a rt ia l ly ov e rlap p ed w ith th a t ofth e bin d in g of isolec t in I B f romG rif f on ia s im p l ic if ol ia I )4. 2 B T h e f u r th e r obse rv a t ion w ith conf oca l la se r sca n n in g m ic roscop e sh ow ed th a t a f ew of N T L I p os t iv e 242. te rm in a ls con ta c ted w ith som e I B la be led te rm in a lsI t w a s f ou n d e lec t ron m ic roscop ica l ly in th e sup e rf ic ia l 2lay e rs of th e sp in a l cord of th e ra t t rea ted w ith su ba ra ch n oid inj ec t ion of cap sa ic in th a t N T L I con ta in in g te r2 . ƒm in a ls con ta c ted w ith d eg en e ra ted te rm in a ls w ith a n d or w ith ou t sy n ap t ic sp ec ia l iz a t ion sT h ese resu l ts in d i2 ca te th a t N T m ay p resy n ap t ica l ly in h ibit th e t ra n sm iss ion of p rim a ry af f e ren t C f ibe rs V ia a x oa x on ic sy n ap s2 ƒ. , es a n d or n on sy n ap t ic con ta c tsA d d it ion a l ly a x od en d r it ic sy n ap ses be tw een d eg en e ra ted te rm in a ls a n d 2.N T L I con ta in in g d en d rites a lso ex is t in th e sp in a l d orsa l h orn 24; Key word s N eu ro ten sin; Imm u no h isto ch em ist ry; C ap sa ic in; G r iffo n ia sim p lic ifo lia IB R a t P r im a ry affe ren t C f ib e r; Sp in a l do r sa l ho rn; () , , iso la ted 2 N eu ro ten sin N T a t r idecap ep t ideo f N T co n ta in in g e lem en t s in th e sp in a l do r sa l 1. , ho rn to th e p r im a ry affe ren t C f ib e r s is no t c lea ro r ig in a lly fo rm th e bo v in e h ypo th a lam u s is , A f te r do r sa l rh izo tom yth e ch an ge o f N T b in d in g w ide ly d ist r ib u ted in th e m amm a lian n e rvo u s sy s2 2, 3 4 , 6. 2tem L o ca liza t io n o f N T reac t ive m a te r ia l site in den sity o r d ist r ib u t io n an d th e in te rac t io n o f 7N TL I te rm in a ls w ith degen e ra ted p r im a ry affe ren t 2an d N T b in d in g site w ith in th e sup e rf ic ia l lam i2 bo u to n s h a s no t b een fo u n d in th e ra t sp in a l do r sa l n ae o f th e sp in a l do r sa l ho rn su gge st s th a t N T n eu2 14 15 . , ho rn H ow eve rZh an g an d h is cow o rk e r sde2 ro n e s co n t r ib u te to sen so ry p ro ce ss a t th e leve l o f tec ted th e m RN A o f N T recep to r in th e do r sa l roo t . th e sp in a l co rdIt h a s b een repo r ted th a t an t ino c i2 () gan g lio n D R G n eu ro n e s o f th e ra t b y u sin g in cep t ive effec t s can b e in du ced b y in t ra th eca l adm in2 situ h yb r id iza t io n an d fo u n d th a t N T recep to r m R 2 ist ra t io n o f N T a t th e lum b a r leve l o f th e sp in a l 8 , 10N A w a s exp re ssed in abo u t 30% o f a ll L 4ƒL 5 D R G 2 an d th a t m ic ro io nco rd o f th e ra t an d m ice , . , n eu ro n e sm a in ly in sm a ll o n e sM o reo ve rth ey re su lt s in ex c ita to 2 top ho re t ic app lica t io n o f N T 13 11, 12 h ave a lso fo u n d N TL I axo axo n ic co n tac t s w ith p r i2 an d in h ib ito ry effec t s o n th e n eu ro n e s in ry .m a ry affe ren t s in th e sp in a l do r sa l ho rn th e ca t an d ra t sp in a l do r sa l ho rn. Imm u no e lec t ro n 5, 6 , In th e p re sen t stu dyb a sed o n se lec t ive b in d2 2m ic ro scop ic stu d ie sh ave show ed th a t N T lik e 24 in g o f IB iso lec t in f rom th e seed s o f G r iffo n ia () imm u no reac t ive N TL Ipo sit ive te rm in a ls fo rm 16sim p lic ifo lia to sm a ll p r im a ry sen so ry n eu ro n e s ƒa symm e t r ica l an do r symm e t r ica l syn ap se s w ith an d sp ec if ic n eu ro to x ic ity o f cap sa ic in to u nm ye li2 ƒ. den d r ite s an do r som a sIt is th e refo re su gge sted 17th a t N T m ay po st syn ap t ica lly ac t o n th e n eu ro n e s , n a ted p r im a ry affe ren t s do u b le f luo re scen ce . , o f th e sp in a l do r sa l ho rnT o da teth e re la t io n sh ip h isto ch em ist ry, co n fo ca l la se r scan n in g m ic ro scop y ( ) an d imm u no e lec t ro n m ic ro scop y a re u sed to ex am 2 210 w a s in se r teda po lye th y len eca th e te r P E 2th ro u gh th e c iste rn a m agn a in to sp in a l su b a rach 2 in e if N TL Ico n ta in in g te rm in a ls a re co n n ec ted no id sp ace o f th e ro st ra l edge o f th e lum b a r en 2 w ith th e cen t ra l te rm in a ls o f p r im a ry affe ren t C () 212la rgem en t L L o f each ra t u sin g a m o d if ica t io n .f ib e r s in th e do r sa l ho rn o f th e ra t sp in a l co rd 18, 19 o f th e m e tho d de sc r ib ed p rev io u sly . Su b se2 M a ter ia ls an d M e thod s () 125% , 20 0ƒqu en t lyΛl o f w vcap sa ic in d isso lved 10 , 2 in a ste r ileo sm o t ica lly b a lan ced so lu t io n co n2 D o u b le f luo re scen ce h isto ch em ist ry o f N TL I an d I () 4 50% ƒB b in d in g an d co n fo ca l la se r scan n in g m ic ro scop y ta in in g vvDM SO w a s in t ra th eca lly in jec t2 . , 250, 300ed v ia th e ca th e te rA f te r th e in jec t io n th e ra t w a s Fo u r m a le W ista r ra t s w e igh in g g 3 , 2a llow ed to su rv ive fo r day srean e sth e t ized 0101ƒw e re p e rfu sed t ran sca rd ia lly w ith m o lL so d i2 , deep ly a s de sc r ib ed abo vean d th en p e rfu sed t ran 2 () (2185% 0ƒ;um p ho sp h a teb u ffe red w vsa lin e PB S ) , 713 , 011ƒsca rd ia lly w ith PB Sfo llow ed w ith PB co n ta in in ig pH th en w ith m o lL p ho sp h a te b u ffe r () () ; 7134% ƒ() () 175% 0PB pH co n ta in in g w vp a rafo rm a lde2 4% ƒƒ, vp a rafo rm a lde lydew vp ic r icw () ƒ1% (lyde an d w vp ic r ic ac id u n de r deep an e sth si2 ) 01075% ƒ. ac id an d w v g lu ta ra ldeh ydeA f te r ( ) a . . , b y in t rap e r ito n ea l in jec t io n ipo f so d ium p en2 th a tth e lum b a r en la rgem en t o f th e sp in a l co rd ( )1100. ƒto b a rb ita l m g k g bo dy w e igh t A f te r p e rfu2 w a s imm ed ia te ly rem o ved an d r in sed in PB S fo r , sio n th e lum b e r en la rgem en t o f th e sp in a l co rd , 2h , an d th en cu t in to c ro ss sec t io n s 50, 70 Λm . w a s imm ed ia te ly rem o ved an d p laced in th e sam e th ick o n a v ib ra tom eT h e sec t io n s w e re in cu b a ted ( ) 11 ?3000 2? 44. ,sequ en t ia lly in rab b it an t iN T se rum fo r hSu b sequ en t ly th e sp ec im en f ix a t ive a t () 4? , 40; 210 35ƒ25% 4?a t fo r hΛgm l b io t in y la ted go a t w a s sa tu ra ted w ith su c ro se in PB S a t 2[ ] an t irab b it IgG an t ibo dy a t room tem p e ra tu re fo r o ve rn igh t. T h e t issu e b lo ck s w e re cu t in to c ro ss ( ) () 2; 31 ? 100 30 hA BC V ec to ra t room tem p e ra2 . sec t io n s Λm th ick o n a f reezin g m ic ro tom eT h e 2. tu re fo r hT h e in cu b a t io n m ed ia w e re p rep a red sec t io n s w e re in cu b a ted a t room tem p e ra tu re se2 ( ) 2% ƒ2b y u sin g PB S co n ta in in g vv no rm a l go a t qu en t ia lly w ith a m ix tu re o f rab b it an t iN T an t i2 () . ?3000; 1 1ƒse rumBo u n d p e ro x ida se w a s v isu a lized b y in cu b a2 se rum In c sta ran d Λgm l b io t in y la ted ( )105 216 ( ) 0ƒ7215 ƒt io n w ith m o lL T r isH C l b u ffe r pH 4 2IB S igm a an d Λgm l f luo re sce in iso th io 2 () () () () 0103% 1003% 0ƒƒ22. co n ta in in g w vDA B an d vvcyan a te F IT C lab e lled av id in D V ec to rfo r h , 20 . 15H 2O 2 a t room tem p e ra tu re fo r m inA f te r T h e in cu b a t io n m ed ia w e re p rep a red b y u sin g PB S ((, , ))3% ƒ2100 2% ƒw a sh in g in PB Sth e sec t io n s w e re o sm ica tedde2 co n ta in in g van d vvT r ito n X v 2.h yd ra ted an d th en f la tem b edded in D u rcup an no rm a l go a t se rum T h e sec t io n s w e re o b se rved (( ) . u n de r a Ze iss ep if luo re scen ce m ic ro scop e A x io 2 F lu k a b e tw een tw o sh ee t s o f p la st icU n de r a , ) (d issec t io n m ic ro scop eth e sup e rf ic ia l laye r s o f th e p ho to w ith an app rop r ia te f ilte r fo r F IT C ex c ita2 , ) , 490 ; 515, 565 do r sa l ho rn w e re rem o vedan d th en cu t in to u lt ra2 450t io n nm em issio n nm o r . ( th in sec t io n sT h e se u lt ra th in sec t io n s w e re do u b le 530, 585 ; ?T ex a s R ed ex c ita t io n nm em issio n co u n te r sta in ed w ith u ran y l ace ta te an d lead c it ra te )615 . 2nm Som e do u b lelab e lled sec t io n s w e re th en 27100 an d th en o b se rved u n de r a H it iach i H e lec t ro n ex am in ed u n de r a co n fo ca l la se r scan n in g m ic ro 2 m ic ro scop e. () 410; scop e L SMZe iss u sin g ex c ita t io n la se r 488 543 b eam s o f nm an d nm w ave len g th s re sp ec2 Re sults t ive ly w ith app rop r ia te em issio n f ilte r s fo r F IT C () ()510, 525 ?570 . nm an d T ex a s R ed nm 24 L igh t m ic ro scop ic B d ist r ib u t io n o f N TL I an d I Imm u no e lec t ro n m ic ro scop y o f N T b in d in g ( ) 200, 250Fo u r m a le W ista r ra t s g w e re In th e t ran sve r se sec t io n s o f th e lum b a r sp in a l , , 2?I 2 u sedan d u n de r th e gen e ra l an e sth e sia w ith so d ico rdN TL I w a s co n cen t ra ted in th e lam in ae () 49 ƒ, . . ,um p en to b a rb ita l m gk g bo dy w e igh tip o f th e do r sa l ho rn an d o b se rved a s tw o d ist in c t () () I . : . 5, 6. b an d F igla o n e b an d ac ro ss lam in a an d th ed r ite s F igo r w ith u n lab e lled som a sSom e ( 2 I ƒ 2 ??th irdo f lam in a lam in aeN TL Ilab e lled den d r ite s w e re a lso seen in lam in ae o u te r o n e ) 2 , 2I ?2 in w h ich N TL Ipo sit ive p ro du c t s w e re a sso 2 bo rde r an d th e o th e r o n e ac ro ss th e in n e r o n e ?? , th ird o f lam in a an d th e o u te r h a lf o f lam in a c ia ted w ith th e m ic ro f ilam en t sm ic ro tu b u le s an d ( )) ƒ?. (7. ? lam in ae bo rde rT h e b an d o f N TL I a t th e . m ito cho n d r ia F ig ƒ?I ƒ? ? bo rde r w a s den se r th an th a t a t th e 2 com po n en t sU lt ra st ru c tu ra l re la t io n sh ip o f N TL I , bo rde ran d th e zo n e b e tw een tw o b an d s exp re ssed w ith th e degen e ra ted te rm in a ls . few imm u no reac t iv it ie sN O N TL Iw a s o b se rved in In th e sp in a l do r sa l ho rn o f th e ra t t rea ted . th e do r so la te ra l t rac t o f L issau e rT h e b in d in g re2 , w ith su b a rach no id in jec t io n o f cap sa ic in m o st o f 24 ac t iv ity o f IB w a s a lso d ist r ib u ted in th e sup e rf i2 2N TL Ico n ta in in g com po n en t s w e re co n n ec ted w ith , c ia l laye r s o f th e sp in a l do r sa l ho rn re st r ic ted in , u n lab e lled den d r ite ssom a s o r te rm in a ls b u t no t ()I 2? . 1. th e lam in ae F igb A ltho u gh th e b in d in g . w ith degen e ra ted te rm in a lsIn som e u lt ra th in sec2 ? reac t iv ity w a s som ew h a t m o re in ten se in lam in a 2 , t io n saxo axo n a l co n tac t s ex isted b e tw een N TL I I , 2th an in lam in a no d ist in c t ive b an dlik e la2 co n ta in in g te rm in a ls an d th e degen e ra ted te rm i2 . ,b e llin g p a t te rn w a s seenIn co n t ra st w ith N TL I , , n a lshow eve rm o st o f th em d id no t show syn ap 2 2′4 th e b in d in g o f IB w a s a lso p re sen t in L issau e rs2, t ic sp ec ia liza t io n. O cca sio n a llyN TL Ipo sit ive .t rac t n e rve te rm in a ls, w h ich co n ta in ed m an y syn ap t ic , U n de r co n fo ca l la se r scan n in g m ic ro scop e, ve sic le sm ade d ist in c t axo axo n a l syn ap se s o n th e m o st o f p re sum ed axo n a l te rm in a ls lab e lled w ith (). 8. degen e ra ted te rm in a ls F igIn add it io n , a few , T ex a s R ed o r F IT C show ed sin g le f luo re scen ceo f axo den d r it ic syn ap se s fo rm ed b y th e degen e ra ted . , th e red o r th e g reenH ow eve ra few o f com po 2 2te rm in a ls an d N TL Ilab e lled den d r ite s w e re fo u n d , n en t s w e re de tec ted to ex h ib it ye llow f luo re scen ce; in th e sam e reg io n s o f th e sp in a l co rdth e se axo 2 ( .th e m ix tu re o f red an d g reen f luo re scen ce F ig (.den d r it ic syn ap se s w e re a lw ay s a symm e t r ica l F ig )2. )9. F ea tu re s o f degen e ra ted te rm in a ls D iscuss ion , U n de r e lec t ro n m ic ro scop eth e sp ec if ic de2 gen e ra ted n e rve te rm in a ls w e re seen in th e sup e rf i2 T h e sup e rf ic ia l lam in ae o f th e sp in a l do r sa l c ia l laye r s o f th e do r sa l ho rn o f th e ra t s t rea ted ho rn a re th e m a in reg io n s o f th e cen t ra l te rm in a2 . w ith cap sa ic inT h e se degen e ra ted te rm in a ls w e re t io n o f u nm ye lin a ted affe ren t s o f C f ib e r s de r ived I 2? . f rom sm a ll sen so ry n eu ro n e s o f th e do r sa l roo t m a in ly fo u n d in lam in ae T h ey d isp layed 20 , gan g lio n . T h e re su lt s in th e p re sen t stu dy b y m a rk ed an d even ly in c rea sin g e lec t ro n den sityo b2 , . . scu re st ru c tu re s o f th e o rgan e lle segir regu la r imm u no f luo re scen ce o b se rva t io n show ed th a t . ve sic le sSom e o f th e degen e ra ted te rm in a ls co n st i2 N TL I w a s re st r ic ted ly d ist r ib u ted in th e sup e rf ic ia l laye r s o f th e sp in a l do r sa l ho rn a s tw o d ist in c t tu ted syn ap t ic g lom e ru li w ith seve ra l su r ro u n d in g )(3, 4. 2 2,ƒ. b an d s o f f ib e ran d te rm in a llik e f luo re scen ce den d r ite s an d o r axo n a l te rm in a ls F ig re2 w h ich w a s co in c iden t w ith th e p rev io u s U lt ra st ru c tu ra l lo ca liza t io n o f N TL I 2, 3, 5 . 2, 2 T h e f luo re scen ce do u b lelab e llin g U n de r e lec t ro n m ic ro scop eN TL I w a s p rom ipo r t sex 2 , , n en t ly p re sen t in axo n s an d axo n a l te rm in a ls am in a t io n revea led th a tin th e sam e reg io n sth e th ro u gho u t th e n eu rop il o f th e sup e rf ic ia l do r sa l d ist r ib u t io n o f N TL I o ve r lapp ed w ith th a t o f th e , 224. 4, ho rn th e po sit ive reac t io n p ro du c t s b e in g m a in ly b in d in g site s fo r IB IB an iso lec t in f rom th e lo ca lized o n th e o u te r su rface s o f sm a ll c lea r ve si2 seed s o f G r iffo n ia sim p lic ifo lia, h a s b een k now n to ( ). 5, 6.c le s an d a lso in la rge den se ve sic le s F ig 2se lec t ive ly b in d to a su b pop u la t io n o f sm a ll d iam e2 T h e se N TL I lab e lled te rm in a ls fo rm ed a symm e t r i2 , te r sen so ry gan g lio n ic n eu ro n e sth e g rea t m a jo r ity 16 . ca l an d symm e t r ica l syn ap se s w ith u n lab e lled den2 o f w h ich supp lie s no c icep to r sT h e ch a rac te r is2 b it io n o f p r im a ry affe ren t n eu ro n e s. T h is h ypo th e2 t ic d ist r ib u t io n o f N TL I an d th e p a r t ia l d ist r ib u t ive 24 ′m a tch b e tw een N TL I an d IB b in d in g reac t iv ity sis h a s b een suppo r ted b y Zh an g an d cow o rk e r s 15st ro n g ly su gge st s th e po ssib le co r re la t io n b e tw een in w h ich th ey h ave dem o n st ra ted th a t w o rk N TL I com po n en t s an d C f ib e r s w h ich in vo lve in sm a ll n eu ro n e s in do r sa l roo t gan g lio n o f th e ra t .2 th e inp u t o f no c icep t io n exp re ssed m RN A fo r N T recep to r an d th a t N TL I T h e fu r th e r o b se rva t io n o f th e f luo re scen ce 2co n ta in in g te rm in a ls co n tac ted no n syn ap t ica lly o n 2th e cen t ra l te rm in a ls o f p r im a ry affe ren t s in th e do u b lelab e lled sec t io n s w ith co n fo ca l la se r scan2 . , sp in a l do r sa l ho rnH ow eve rth ey d id no t iden t ify n in g m ic ro scop e iden t if ied som e e lem en t s w ith ye l2 w h e th e r th e p r im a ry affe ren t s th e te rm in a ls o f low f luo re scen ce fo rm ed b y th e m ix in g o f red an d 2w h ich w e re co n tac ted o n b y N TL Ipo sit ive te rm i2 . 2g reen f luo re scen ceN T co n ta in in g axo n s in th e . n a ls w e re C f ib e r s in th e ir stu dyIt h a s b een o b2 sp in a l do r sa l ho rn w a s tho u gh t to b e o r ig in a ted no t se rved in o u r co n fo ca l la se r scan n in g m c iro scop ic f rom p r im a ry affe ren t n eu ro n e s b u t f rom in t r in sic , 2 n eu ro n e ssin ce o n th e co n d it io n o f do r sa l rh izo to 2 an d imm u no e lec t ro n m ic ro scop ic w o rk th a t N TL I, , 2 m yt rea tm en t w ith cap sa ic in o n n eo n a ta l an im a lspo sit ive axo n ic te rm in a ls no t o n ly fo rm ed no n , o r sp in a l h em isec t io n no de tec tab le dep le t io n o f syn ap t ic axo axo n ic co n tac t s b u t a lso fo rm ed def i2 14, 24, 25 .N T w a s cau sed in th e sp in a l do r sa l ho rn n ite axo axo n ic syn ap se s w ith C f ib e r te rm in a ls , T h u sth e m ix ed f luo re scen ce o b se rved in th e p re2 24w h ich w e re iden t if ied b y th e se lec t iv ity o f IB ’ s sen t stu dy h in t s th e po ssib ility th a t th e re a re c lo se b in d in g to C f ib e r an d th e sp ec if ic ity o f cap sa ic in ’ s 2, co n tac t s b e tw een N TL Ico n ta in in g axo n ic te rm i2 en dam ag in g C f ib e rw h ich fu r th e r p ro v ided m o re m o rp ho lo g ica l ev iden ce s fo r th e po ssib ility th a t N T n a ls an d cen t ra l te rm in a ls o f C f ib e r s iden t if ied b y 224. IB b in d in g reac t iv ityW h e th e r th e co n tac t s a re m igh t h ave p re syn ap t ica lly in h ib ito ry effec t o n p r i2 syn ap se s o r no t is st ill u n ce r ta in ow in g to th e lim i2 ƒ m a ry affe ren t C f ib e r s v ia axo axo n ic syn ap se s an dta t io n o f th e re so lu t io n o f co n fo ca l la se r scan n in g 2. o r no n syn ap t ic axo axo n ic co n tac t sT h e m o st o f .m ic ro scop e axo axo n ic co n tac t s fo u n d in o u r w o rk d id no t show , It h a s b een gen e ra lly accep ted th a t cap sa ic in syn ap t ic sp ec ia liza t io n th e typ ica l syn ap se s w e re ex e r t s a sp ec if ic n eu ro to x ic ac t io n o n C f ib e r s o f ve ry few. In fac t, it is lik e ly th a t n eu rop ep t ide s 17 15 . 2.p r im a ry affe ren t sCo n sisten t w ith th e p rev io u s d iffu sed no n syn ap t ica lly a re re lea sed an d 17, 23, 24 , it w a s o b se rved u n de r e lec t ro n m i2 stu d ie sT h u s, N T in th e sp in a l do r sa l ho rn m ay b e m a in ly 2c ro scop e th a t in t ra th eca l in jec t io n o f cap sa ic in in no n syn ap t ica lly re lea sed to ex e r t a d irec t ac t io n o n .adu lt ra t se lec t ive ly in du ced degen e ra t io n o f p r im a2 p r im a ry C affe ren t s 5, 6. ry affe ren t C f ib e r sT h e te rm in a ls o f degen e ra ted , th e a symm e t2 S im ila r to th e p r io r repo r t s p r im a ry affe ren t C f ib e r s in th e su b stan t ia ge la t i2 r ica l axo den d r it ic syn ap se s b e tw een N TL I axo n ic no sa app ea red ch a rac te r ist ic e lec t ro n m ic ro scop ic te rm in a ls an d u n lab e lled den d r ite s a re m o re f re2 , im age sd isp a ly in g m a rk ed h igh e lec t ro n den sity qu en t ly seen th an symm e t r ica l o n e s in th e p re sen t an d o f ten fo rm in g syn ap t ic g lom e ru li w ith su r2 . stu dyT h e a symm e t r ica l axo den d r it ic syn ap se s a re . . ro u n d in g den d r ite sT h e o b se rva t io n th a t th e cen2 ex c ita to ryN T is m o re lik e ly to b e an ex c ita to ry , n eu rop ep t idew h ich h a s b een in d ica ted b y th e p re2 t ra l te rm in a l bo u to n s o f u nm ye lin a ted o r th in 25 v io u s e lec t rop h y sio lo g ica l f in d in g s th a t N T p re2 m ye lin a ted p r im a ry affe ren t sis fu r th e r suppo r t2 dom in an t ly ex e r t s ex c ita to ry effec t s o n th e sp in a l . ed b y o u r da taIn t ra th eca l adm in ist ra t io n o f N T 11, 12 , do r sa l n eu ro n e san d is fu r th e r suppo r ted b y re su lted in an a lge sia in a va r ie ty o f b eh av io ra l a s2 8 , 10, 2226, 27 [] 2o u r re su lt th a t th e a symm e t r ic axo N TL I den2 say sb u t d id no t b lo ck p a in b eh av io r in 2 ( ) d r it ic u n lco b e lled syn ap se s a re m o re o f ten o b2 du ced b y in t ra th eca l su b stan ce P , a n eu rop ep t ide in vo lved in no c icep t io n. A s m en t io n ed It is th e refo re po ssib le se rved th an a symm e t r ic o n e s. th a t N T an a lge sia m ay o ccu r v ia p re syn ap t ic in h i2 abo ve, N T h a s app a ren t an a lge sia a t th e leve l o f . 6 Seybo ld V S and M a ley BU lt ra st ruc tu ra l study o f neu2 th e sp in a l co rd an d th e axo axo n ic syn ap t ic an d ro ten sin imm uno reac t iv ity in th e sup e rf ic ia l lam inae o f 2 co n tac t s a re ra re, th e po st syn ap t ic no n syn ap t ic th e do r sa l ho rn o f th e ra t. P ep t ide s 1984; 5: 117921189 is th e refo re lik e ly to b e m o re im po r2 ac t io n o f N T . 7 Yo ung W S and Kuh a r M JN eu ro ten sin recep to r lo ca l2 . tan t th an it s p re syn ap t ic o n eIf th e ex c ita to ry ef2 fec t o f N T o n th e sp in a l do r sa l n eu ro n e s is in vo lved iza t io n by ligh t m ic ro scop ic au to rad io g rap h y in th e ra t , in an a lge siath e an a lge sia w o u ld b e m ed ia ted v ia . 1981; 206: 2732285b ra inB ra in R e s .an in h ib ito ry in te rn eu ro n 8 H y lde r JL K and W ilco x GL. A n t ino c icep t ive ac t io n o f , A dd it io n a llyit w a s fo u n d th a t N TL I w a s a l2 2. 1983; 4: 517in t ra th eca l neu ro ten sin in m iceP ep t ide s so lo ca lized w ith in som e den d r ite s in th e sp in a l 520 , , 2do r sa l ho rn m o reo ve rsom e N TL Ico n ta in in g , . 9 H an SP X ie GX and H an J SInvo lvem en t o f dyno rp h in den d r it ic p ro f ile s w e re show n to b e po st syn ap t ic to 2in th e an t ino c icep t io n induced by in t ra th eca l in jec t io n o f . th e degen e ra ted te rm in a lsT h is typ e o f syn ap t ic neu ro ten sin in th e ra t. C h ine se A c ta P h y sio lo g ica S in ica co n n ec t io n m ay b e re la ted to th e in c rea se o f N T re2 ( )() 1987; 39 1: 19225 in C h ine se lea se in re spo n se to p e r ip h e ra l n e rve st im u la2 , , , . 2 10 Sp am p ina to SR om ua ld P C anda le t t i Se t a lD ist in 25 . t io n , gu ish ab le effec t s o f in t ra th eca l dyno rp h in ssom a to 2 2, sta t inneu ro ten sin and sca lc ito n in o n no c icep t io n and A ckn owledgm en ts . 1988; 35: 952104m o to r func t io n in th e ra tP a in T h e au tho r s a re g ra tefu l fo r th e p ho to g rap h ic , 11 M ile t ic V and R and ic M N eu ro ten sin exc ite s ca t sp ina l . h e lp o f M rX u e ju n K an g a t D ep a r tm en t o f H isto l2 2 . .?1979; I neu ro n s lo ca ted in lam inae B ra in R e s , , o gy an d Em b ryo lo gyT o n g ji M ed ica l U n ive r sity169: 6002604 . . . PRo f C h in aT h is w o rk w a s suppo r ted in p a r t 12 S tanzio ne P and Z ieg lgan sbe rgen W . A c t io n o f b y N a t io n a l N a tu ra l Sc ien ce Fo u n da t io n o f C h in a . ne ro ten sin o n sp ina l co rd neu ro n s in th e ra tB ra in ()39370250. 21181993; 268: 111R e s 收稿 1998212 修回 1999208 13 . H en ry JLE lec t rop h y sio lo g ica l stud ie s o n th e neu ro ac2 () 本文图 1- 8 见插图第 8 页 . , t ive p rop e r t ie s o f N eu ro ten sinN eu ro ten sina b ra in Ref eren ce s . 1982;and ga st ro in te st ina l p ep t ideA nn N Y A cad Sc i 400: 2162227 1 C a r raw ay R E and L eem an SE. T h e iso la t io n o f a new , . N iko v ic M H un t SP and Ke lly J SE ffec t o f do r sa l rh i2 14 , , h ypo ten sive p ep t ideneu ro ten sinf rom bo v ine h ypo th a2 zo tom y o n au to rad io g rop h ic d ist r ibu t io n o f op ia te and 26861. 1973; 248: 6854lam iJ B io l C h em 2neu ro ten sin recep to r s and neu ro ten sinlik e imm uno re2 2 , . Ko baya sh i RM B row n M R and P e r r in M HR eg io na l . 1981; 230:ac t iv ity w ith in th e ra t sp ina l co rdB ra in R e s d ist r ibu t io n o f neu ro ten sin and som a to sta t in in ra t 1112119 126: 5842588 b ra in. B ra in R e s 1977; , 2, , . Zh ang X X u ZQ B ao L e t a lCom p lem en ta ry d is2 . : 215 3 U h l GR and Suyde r SHN eu ro ten sinimm uno h is t r ibu t io n o f recep to r s fo r neu ro ten sin and N P Y in sm a ll .to ch em ica l lo ca liza t io n in ra t cen t ra l ne rvo u s sy stem neu ro n s in ra t lum ba r D R G s and regu la t io n o f th e re2 74: 405924063 P ro c N a t A cad Sc i U SA 1977; . cep to r s and p ep t ide s af te r p e r ip h e ra l axo tom yJ N eu2 4 Seybo ld V S and E lde R P. N eu ro ten sin imm uno reac t iv ity () 4: 273322747 : ro sc i 1995; 15 in th e sup e rf ic ia l lam inae o f th e do r sa l ho rn o f th e ra tI L igh t m ic ro scop ic stud ie s o f ce ll bo d ie s and p ro x im a l . 16 S ilve rm an JD and K ruge r LSe lec t ive neu ro na l g lyco 2 . 1982; 205: 892100dend r ite sJ Com p N eu ro l : co n juga te exp re ssio n in sen so ry and au to nom ic gang liaD if ig lia M , A ro n in N and L eem an SE. U lt ra st ruc tu ra l 5 re la t io n o f lec t in reac t iv ity to p ep t ide and enzym e . 1990; 19: 7892801lo ca liza t io n o f imm uno reac t ive neu ro ten sin in th e m o n2 m a rk e r sJ N eu ro cy to l 2 17 225: 1k ey sup e rf ic ia l do r sa l ho rn. Com p N eu ro l 1984; F itzge ra ld M . C ap sa ic in and sen so ry neu ro ne s2A R e2 12 v iew. P a in 1983; 15: 1092130 2243 1982; 400: 22818 Yak sh TL and R udy TA. C h ro n ic ca th e te r iza t io n o f th e , . P a le rm o N N B row n H K and Sm ith DLSe lec t ive neu2 17: 23 sp ina l suba rach no id sp ace. P h y sio l B eh av 1976; 2ro to x ic ac t io n o f cap sa ic in o n g lom e ru la r C typ e te rm i2 103121036 . 1981; 208;19 na ls in ra t sub stan t ia ge la t io n saB ra in R e s L i H and M izuno N. A n imm uno e lec t ro n m ic ro scop ic 506- 510 22study o n th e re la t io n s o f m e tenk ep h a linlabe lled and - - . un labe lled e lem en t s to C f ibe r in th e do r sa l ho rn o f th e 24 R ibe iro da silva A and Com im b ra AC ap sa ic in () . 1996; 12 4:cau se s se lec t ive dam age to typ e I synap t ic g lom e ru li in ra t sp ina l co rdC h ine se J N eu ro ana tom y ( . 1984, 290: 380-)4012406 in C h ine se ra t sub stan t ia ge la t ino saB ra in R e s 383 20 Sug iu ra Y , L ee CR and P e r l ER. C en t ra l p ro jec t io n s o f 22. R ibe iro da S ilva A and Com im b ra ATw o typ e s o f () 25 iden t if ied unm ye lina ted C affe ren t f ibe r s inne rva t ing I 2synap t ic g lom e ru li and th e ir d ist r ibu t io n in lam inae m amm a liam sk in. Sc ience 1986; 34: 2582361 , , , . 221 ? 209: J anc so GH o k fe lt T L undbe rg JM e t a lImm uno o f th e ra t sp ina l co rd. J Com p N eu ro l 1982, 176- 186 gōàh isto ch em ica l stud ie s o n th e effec t o f cap sa ic in o n sp 26 . ina l and m edu lla ry p ep t ide and m o no am ine neu ro n s u H y lden JL K and W ilco x G lSubcu taneo u s h yp e r to n ic , , , sing an t ise ra to sub stance P ga st r in CC K som a to 2 sa line a ssay a s a m ea su re o f ana lge sia fo llow ing in2 2, , , 5, : sta t in V IP enk ep h a linneu ro ten sin and h yd ro x2 t ra th eca l neu ro ten sinop io id s and m o no am ine in m icey t ryp tam ine. J N eu ro cy to l 1981; 210: 9632980 co r re la t io n w ith inh ib it io n o f sub stance P induced be2 , , , . 22 8: 985 Yak sh TL Schm au ss C M icevych P E e t a lP h a rm a2 h av io r. So c N eu ro sc i A b st 1982, , , 27 Seybo ld V S, H y lden JL K and W ilco x GL. In t ra th eca l co lo g ica l stud ie s o n th e app lica t io nd ispo sit io nand . ,re lea se o f neu ro ten sin in th e sp ina l co rdN eu ro ten sin : sub stance P and som a to sta t in in ra t sB eh av io r s ind ica2 . a b ra in and ga st ro in te st ina l p ep t ideA nn N Y A cad Sc i t ive o f sen sa t io n p ep t ide s, 1982, 3: 49254 大鼠脊髓后角内神经降压素对一级传入 纤维突触前 C 抑制效应的形态学证据 李 和 张亦农 张敏海 杨世明 李宏莲 ()同济医科大学组织与胚胎学教研室, 武汉 430030 本研究的目的是为了揭示大鼠脊髓后角内的神经降压素是否对一级传入 C 纤维进行突触前调节。荧光显微镜下观察到, () ()在脊髓I, ? 层内,神经降压素样免疫反应性 的分布与来源于西非单豆素的同工凝集素I 2I 2的结合位点4 N TL IB B 到分布有部分重叠。在共聚焦激光扫描显微镜下进一步观察显微镜下, 在蛛网膜下腔注射辣椒素的大鼠脊髓后角浅层内, 一 些 阳性终末与变性终末形成突触性和ƒ或非突触性接触。以上结果明, 可通过轴轴突触和ƒ或非突触性轴轴接触 N TL I N T 的一级传入 纤维的传入进行突触前抑制。 此外, 脊髓后角内还存在变性终末与含 树突形成的轴树突触。C N TL I 辣椒素脊髓后角 大鼠一级传入 C 纤维 关键词 神经降压素 免疫组织化学 西非单豆素 2 4; IB () 2, 4 th e lum ba r co rdshow ing N TL I aand IB b ind2 EXPL A NA T IO N O F F IGURES () I 2? . ing b in th e lam inae o f th e do r sa l ho rnand F ig. 1 F luo re scence p ho tom ic ro g rap h s o f a c ro ss sec t io n o f , b w e re tak en f rom th e sam e sec t io nth e a ste r isk s 2po sit ive p ro duc t s a re d is2 N o te th a t th e N TL Iin a and b ind ica t ing th e iden t ica l p ro f ile o f a b loo d 2. ve sse lN o te th e p a r t ia l o ve r lap o f N TL Iw ith th e I t r ibu ted in bo th sm a ll c lea r ve sic le s and la rge den se ()4 .B b ind ing reac t iv ity ve sic le s . L () 2 6 F ig. P ho tom ic ro g rap h tak en w ith a co nfo ca l la se r scan2 F ig. A n axo na l te rm ina l w ith N TL I N T afo rm s a sym 2 2(), n ing m ic ro scop eshow ing N TL Ipo sit ive te rm ina ls m e t r ic synap se w ith an un labe led dend r ite D in ( ) ( ) 224red and IB labe led te rm ina ls g reen a t lam i2 th e sup e rf ic ia llaye r s o f th e lum ba r sp ina l do r sa l ƒ?. ? nae bo rde rP re sum ed axo axo na l co n tac t s . ho rnT h e a r row s po in t to th e po st synap t ic sp ec ia l2 () .ye llow a re ind ica ted by comm a s .iza t io n E lec t ro n m ic ro scop ic p ho to g rap h o f th e sup e rf ic ia l () 3 7 F ig. F ig. A dend r it ic p ro f ile co n ta in ing N TL I N T d in th e , 3laye r s o f th e lum ba r sp ina l do r sa l ho rnshow ing .sup e rf ic ia l laye r s o f th e lum ba r sp ina l do r sa l ho rn () A n axo na l te rm ina l co n ta in ing N TL I N T asynap s2 ()8 F ig. d isp lay ing m a rk ed ly degene ra ted te rm ina ls D eg () .e s o n a degene ra ted te rm ina l D egin th e sup e rf ic ia l inc rea sed e lec t ro n den sity .laye r s o f th e lum ba r sp ina l do r sa l ho rn () 4 A degene ra ted te rm ina l D eg fo rm s a synap t ic F ig. A n a symm e t r ic axo dend r it ic synap se be tw een a de2 () 3 1232 9 F ig. g lum e ru lu s w ith dend r ite s D D and axo na l () gene ra ted te rm ina l D eg and a dend r it ic p ro f ile () 1, 2te rm ina ls A A in th e sup e rf ic ia l laye r s o f th e () w ith N TL I N T d in th e sup e rf ic ia l laye r s o f th e .lum ba r sp ina l do r sa l ho rn .lum ba r sp ina l do r sa l ho rn 5 ()F ig. 22 synap se s a sym A N TL Ipo sit ive te rm ina l N T a , 100 15. 1, 5 . 2 0Sca le ba rΛm in F igΛm in F igand () m e t r ica lly o n an un labe led dend r ite D in th e su2 29.3Λm in F ig s . p e rf ic ia l laye r s o f th e lum ba r sp ina l do r sa l ho rn .T h e a r row s po in t to th e po st synap t ic sp ec iliza t io n 李和等 1 大鼠脊髓后角内神经降压素对一级传入 C 纤维突触前抑制效应的形态学证据 () 正文见第 229 页, 图说明见正文后第 235 页 ?8?