GDNF 220YL) 2,52. (glial cell line2de2. 015d 15d rived neurotrophic factor GDNF) 1993 Lin B49 (L15) (FBS) (poly2l2lysin, pll) (laminin, LN) GDNF GDNF

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1 (6) Basic Medical Sciences and Clinics 625 : ( 2003) GDNF 3,,,,, (, ) : ( GDNF) (DRGn), MTT 1 g/ L 10 g/ L 50 g/ L 100 g/ L GDNF : GDNF GDNF : ; ; :R32912 :A (glial cell line2de2, 015d 15d rived neurotrophic factor GDNF) 1993 Lin B49 (FBS) B27 Leibovitzπs (L15), 2 Gibco ; L2 (L2glutamine ), GDNF (poly2l2lysin, pll) (laminin, LN) ( trypsin) Sigma ; GDNF GDNF Biotech (2), (neuron specific enolase NSE) (, ), SABC (, GDNF ) (3) MTT[32(4,52 220YL) 2,52 ] Sigma, GDNF ( dorsal root 112 DRGn ganglion neurons DRGn), E15, [1 ] DRGn, DRGn, 0125 %, GDNF 37 20min, (FBS), DMEM 1 111, 50min, L - 1, PLL (011g/ L) LN(20mg/ L) :, 5 % CO 2 (37, ) 24h SD ( ), : : : (302515) 3

2 626 Basic Medical Sciences and Clinics (6) 113 Neurobasal, 6g/ L D2, 2mmol/ L, 1 % FBS, 20mL/ L B27 5 : (1) GDNF ; (2) 4 GDNF, 1 g/ L,10 g/ L,50 g/ L,100 g/ L 2, d 4d 7d DRGn, 1 GDNF DRGn Fig 1 The effects of GDNF on the survival 10,, ( 2 ) NSE DRGn 115 MTT DRGn 96 2d 4d 7d, 10 mol/ L MTT, 4h,, 100 L (DMSO), MTT, Rosys anthos 2010 OD, 490nm 620nm 116 SPSS, ( x s), t GDNF DRGn 12h 18h, GDNF 2d 3d,, 2 GDNF DRGn Fig 2 The effects of GDNF on the neurite length ; 4d 5d,, 3 GDNF DRGn Fig 3 The effects of GDNF on the neurite outgrowth,, GDNF,1 g/ L 100 g/ L, 212 GDNF DRGn ( P < 0105) 2d 3d,,,4d 5d,,, GDNF, 4d 5d ( 1,5)

3 (6) Basic Medical Sciences and Clinics 627,,, GDNF,1 g/ L 50 g/ L ( P < 0101) 1 g/ L GDNF 10 g/ L GDNF,50 g/ L GDNF 750 m, 100 g/ L GDNF, 10 g/ L GDNF ( 2,3,5) 213 GDNF DRGn MTT OD 4 GDNF DRGn Fig 4 The effects of GDNF on cultured DRGn( MTT),,OD,, 2d 4d 7d min, 2 OD,0 g/ L GDNF GDNF, 1 g/ L 50 g/ L,,,OD ( P < 0105), DRGn ( 4), DRGn, 3 GDNF GDNF neurturi persephin arte2 GDNF DRGn 5 GDNF DRGn Fig 5 Effects of different GDNF on dissociated DRGn cultured for 4 days. phase contrast microscopy( 200) ; A. 0 g/ L ; B. 1 g/ L ; C. 10 g/ L ; D. 50 g/ L ; E. 100 g/ L

4 628 Basic Medical Sciences and Clinics (6) [5 DRG 50min, ] Neurobasal,, GDNF GDNF GDNF 1 g/ L GDNF,, ( GPI), GDNF GFR ( GFR 1 GDNF, GFR 2 GFR 3 GFR 4) c2ret,, GDNF GFR 1, GDNF DRGn c2ret,,, DRGn, 100 g/ L GDNF DRGn Ret GFR 1,,, GDNF, 50 g/ L GDNF, DRGn Ret GFR 1, trk mr2 GDNF, NA, NGF, GDNF,, GFR 1 [6 ], Forgie A [7 ] RT2PCR, DRG GDNF mrnas, DRGn Ret mrna, GDNF GDNF GFR 1, DRGn, GDNF GDNF DRGn GDNF GDNF [2 DRGn ] GDNF DRGn, IB4 DRG GDNF, GDNF [3 ] GDNF [4 ], : [1 ],,,. GDNF [J ]. J Neurosci, 2000,20 (1) : [J ]. [ 5 ] Gavazzi, Kumar RD, McMahon SB, et al. Growth responses of, 2000,5 : [2 ] Moore MW, Klein RD, Farinas I, et al. Renal and neuronal abnormalities in mice lacking GDNF[J ]. Nature,1996, 382 : [3 ] Buj2Bello A, Buchman VL, Horton A, et al. GDNF is an age2 specific survival factor for sensory and autonomic neurons[j ]. Neuron, 1995, 15 : [4 ] Bennett DL, Boucher TJ, Armanin MP, et al. The glial cell lin2derived neurotrophic factor family receptor components are differentially regulated within sensory neurons after nerve injury different subpopulations of adult sensory neurons to neurotrophic factors in vitro[j ]. Eur J Neurosci, 1999, 11 : [6 ] Boucher TJ, Okuse K, Bennett DL. Potent analgesic effects of GDNF in neuropathic pain states[j ]. Science, 2000, 290 : [7 ] Forgie A,Doxakis E, Buj2Bello A, et al. Differences and de2 velopmental changes in the responsiveness of PNS neurons to GDNF and neurturin[j ]. Mol Cell Neurosci, 1999, 13(6) :

5 (6) Basic Medical Sciences and Clinics 629 Effects of GDNF on survival and axon growth of dorsal root ganglion neurons of rat in vitro WANG Xiao2juan, GUO Li, WANG Li2qin, et al. (Department of Neurology, the Second Hospital, HeBei Medical University, Shijiazhuang , China) Abstract : To explore the effects of glial cell line2derived neurotrophic factor on the survival and neurite growth of dorsal root ganglion neurons(drgn) from normal rat embryo. DRGn dissociated cell culture system from rat embryo was estab2 lished by the primary dissociated culture methods of nerve tissue. The survival and growth of DRGn cultured in NB1 con2 taining 1 g/ L 10 g/ L 50 g/ L and 100 g/ L GDNF respectively were observed in cytomorphology and by MTT method. The results showed that GDNF can significantly enhance the survival and neurite outgrowth of normal rat sensory neurons. This indicated that GDNF can exert a neurotrophic action on developmental sensory neurons from normal rat embryo. Key words : glial cell line2derived neurotrophic factor ; dorsal root ganglion ; cell culture corresponding author : LI Chun2yan ( 624 ), 2, 26 ( IL26), (TNF2 ) C,,, C, 26 alpha BIO1COM(2003/ 10/ 31) , 24 24, 10 %, 2,,, Neurology,2003,61 : , ,,,,,,, BIO1COM (2003/ 11/ 13), Temple University,, WMSD(work2 related musculoskeletal disorders), (carpal tunnel syndrome) BIO1COM(2003/ 11/ 13) ( 633 )