Original Articles

Neurochemical Research

, Volume 21, Issue 5, pp 619-627

First online:

Characterization and transplantation of two neuronal cell lines with dopaminergic properties

  • Frank S. AdamsAffiliated withDepartment of Medicine, University of Colorado Health Sciences Center
  • , Francisco G. La RosaAffiliated withDepartment of Pathology, School of Medicine, University of Colorado Health Sciences CenterCenter for Vitamins and Cancer Research, Dept. of Radiology, University of Colorado Health Sciences CenterThe Webb-Waring Institute, University of Colorado Health Sciences Center
  • , Sanjay KumarAffiliated withCenter for Vitamins and Cancer Research, Dept. of Radiology, University of Colorado Health Sciences Center
  • , Judith Edwards-PrasadAffiliated withCenter for Vitamins and Cancer Research, Dept. of Radiology, University of Colorado Health Sciences Center
  • , Susan KentrotiAffiliated withDepartment of Pharmacology and Psychiatry, University of Colorado Health Sciences Center
  • , Antonia VernadakisAffiliated withDepartment of Pharmacology and Psychiatry, University of Colorado Health Sciences Center
  • , Curt R. FreedAffiliated withDepartment of Medicine, University of Colorado Health Sciences Center
  • , Kedar N. PrasadAffiliated withCenter for Vitamins and Cancer Research, Dept. of Radiology, University of Colorado Health Sciences Center

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Abstract

Immortalized rat mesencephalic cells (1RB3AN27) produced dopamine (DA) at a level that was higher than produced by undifferentiated or differentiated murine neuroblastoma cells (NBP2) in culture. Treatment of 1RB3AN27 and NBP2 cells with a cAMP stimulating agent increased tyrosine hydroxylase (TH) activity and the intensity of immunostaining for the DA transporter protein (DAT). 1RB3AN27 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). 1RB3AN27 cells exhibited β- and α-adrenoreceptors, and prostaglandin E1 receptors, all of which were linked to adenylate cyclase (AC). Dopamine receptor (D1) and cholinergic muscarinic receptors linked to AC were not detectable. The levels of PKCα and PKCβ isoforms were higher than those of PKCγ and PKCδ in 1RB3AN27 cells. The 1RB3AN27 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 NBP2 cells. The grafted 1RB3AN27 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.

Key words

Immortalization SV40 large T-antigen neuroblastoma rotational behavior cyclic AMP tyrosine hydroxylase