Characterization and transplantation of two neuronal cell lines with dopaminergic properties

被引:54
作者
Adams, FS
LaRosa, FG
Kumar, S
EdwardsPrasad, J
Kentroti, S
Vernadakis, A
Freed, CR
Prasad, KN
机构
[1] UNIV COLORADO,HLTH SCI CTR,DEPT RADIOL,CTR VITAMINS & CANC RES,DENVER,CO 80262
[2] UNIV COLORADO,HLTH SCI CTR,DEPT MED,DENVER,CO 80262
[3] UNIV COLORADO,HLTH SCI CTR,DEPT PHARMACOL & PSYCHIAT,DENVER,CO 80262
[4] UNIV COLORADO,HLTH SCI CTR,DEPT PATHOL,DENVER,CO 80262
[5] UNIV COLORADO,HLTH SCI CTR,SCH MED,DENVER,CO 80262
[6] UNIV COLORADO,HLTH SCI CTR,WEBBING WARING INST,DENVER,CO 80262
关键词
immortalization; SV40 large T-antigen; neuroblastoma; rotational behavior; cyclic AMP; tyrosine hydroxylase;
D O I
10.1007/BF02527762
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Immortalized rat mesencephalic cells (IRB(3)AN(27)) produced dopamine (DA) at a level that was higher than produced by undifferentiated or differentiated murine neuroblastoma cells (NBP2) in culture. Treatment of IRB(3)AN(27) and NBP2 cells with a cAMP stimulating agent increased tyrosine hydroxylase (TH) activity and the intensity of immunostaining for the DA transporter protein (DAT). IRB(3)AN(27) cells were labelled with primary antibodies to neuron specific enolase (NSE) and nestin and exhibited very little or no labeling with anti-glial fibrillary acidic protein (GFAP). IRB(3)AN(27) cells exhibited beta- and alpha-adrenoreceptors, and prostaglandin E(1) receptors, all of which were linked to adenylate cyclase (AC). Dopamine receptor (D-1) and cholinergic muscarinic receptors linked to AC were not detectable. The levels of PKC alpha and PKC beta isoforms were higher than those of PKC gamma and PKC delta in IRB(3)AN(27) cells. The IRB(3)AN(27) cells were more effective in reducing the rate of methamphetamine-induced turning in rats with unilateral 6-OHDA lesion of the nigrostriatal system than differentiated NBP, cells. The grafted IRB(3)AN(27) were viable as determined by DiI labelling, but they did not divide and did not produce T-antigen protein; however, when these grafted cells were cultured in vitro, they resumed production of T-antigen and proliferated after the primary glia cells and neurons of host brain died due to maturation and subsequent degeneration. Examination of H&E stained sections of the grafted sites revealed no evidence of infiltration of inflammatory cells in the grafted area suggesting that these cells were not immunogenic. They also did not form tumors.
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页码:619 / 627
页数:9
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