A novel N18TG2 x mesencephalon cell hybrid expresses properties that suggest a dopaminergic cell line of substantia nigra origin

A dopaminergic neuroblastoma was derived using somatic cell fusion of rat embryonic mesencephalon cells and the murine neuroblastoma-glioma cell line N18TG2. The resulting interspecies hybrid, named MES23.5, has retained a stable phenotype and karyotype for a continuous culture period of 1 year. The hybrid exhibits several properties that suggest that the parent primary neurons originated in the substantia nigra. The cell line contains tyrosine hydroxylase, which is identifiable both by biochemical and immunological methods and synthesizes dopamine, but no other catecholamine. Additionally, the cell line expresses apparent voltage-gated CA2+ channels as measured by high-affinity omega- conotoxin binding. The MES23.5 omega-conotoxin receptors are of similar affinity class to those found in adult rat mesencephalon. No dihydropyridine receptors, as measured by PN200–100 ligand binding, are present. None of these properties are found in the N18TG2 parent. At least three neuronal features, namely, tyrosine hydroxylase, dopamine synthesis, and omega-conotoxin receptor expression, are quantitatively elevated after sustained treatment with cAMP analogs. The cell line expresses a complex range of neural properties found in the dopaminergic neurons of the substantia nigra, and may therefore be useful elucidating further details of their cell biology.

[1]  S. Gruber,et al.  The epidemiology of Parkinson's disease. A case-control study of young-onset and old-onset patients. , 1991 .

[2]  A. Ludolph,et al.  Slow toxins, biologic markers, and long‐latency neurodegenerative disease in the western Pacific region , 1991, Neurology.

[3]  M. M. Usowicz,et al.  Differential expression by nerve growth factor of two types of Ca2+ channels in rat phaeochromocytoma cell lines. , 1990, The Journal of physiology.

[4]  E. Lewis,et al.  Interaction of cyclic AMP and cell-cell contact in the control of tyrosine hydroxylase RNA. , 1990, Brain research. Molecular brain research.

[5]  B. Wainer,et al.  Development and characterization of clonal cell lines derived from septal cholinergic neurons , 1990, Brain Research.

[6]  R. McKay,et al.  The use of cell lines in neurobiology , 1990, Trends in Neurosciences.

[7]  M. Tohyama,et al.  Distribution of the ω-conotoxin receptor in rat brain. An autoradiographic mapping , 1989, Neuroscience.

[8]  A. Dwork,et al.  Biochemical properties of monoamine-rich human neuroblastoma cells , 1989, Brain Research.

[9]  C. Lévêque,et al.  Characterization of the omega-conotoxin-binding molecule in rat brain synaptosomes and cultured neurons. , 1988, Molecular pharmacology.

[10]  A. L. Cahill,et al.  Preganglionic stimulation increases the phosphorylation of tyrosine hydroxylase in the superior cervical ganglion by both cAMP-dependent and Ca2+-dependent protein kinases. , 1987, Biochimica et biophysica acta.

[11]  E. Sher,et al.  Cholinergic receptors, ion channels, neurotransmitter synthesis, and neurite outgrowth are independently regulated during the in vitro differentiation of a human neuroblastoma cell line. , 1987, Differentiation; research in biological diversity.

[12]  R. Roskoski,et al.  Activation of Tyrosine Hydroxylase in PC12 Cells by the Cyclic GMP and Cyclic AMP Second Messenger Systems , 1987, Journal of neurochemistry.

[13]  B. Wainer,et al.  Neuronal properties of clonal hybrid cell lines derived from central cholinergic neurons. , 1986, Science.

[14]  P. Curella,et al.  Induction of mRNA for tyrosine hydroxylase by cyclic AMP and glucocorticoids in a rat pheochromocytoma cell line: evidence for the regulation of tyrosine hydroxylase synthesis by multiple mechanisms in cells exposed to elevated levels of both inducing agents. , 1986, Molecular pharmacology.

[15]  A. Olivier,et al.  Comparative autoradiographic distribution of calcium channel antagonist binding sites for 1,4-dihydropyridine and phenylalkylamine in rat, guinea pig and human brain , 1985, Progress in Neuro-Psychopharmacology and Biological Psychiatry.

[16]  P. K. Smith,et al.  Measurement of protein using bicinchoninic acid. , 1985, Analytical biochemistry.

[17]  K. Misono,et al.  Purification and characterization of rat striatal tyrosine hydroxylase. Comparison of the activation by cyclic AMP-dependent phosphorylation and by other effectors. , 1985, The Journal of biological chemistry.

[18]  H. Pakkenberg,et al.  The clinical syndrome of striatal dopamine deficiency. Parkinsonism induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). , 1985, The New England journal of medicine.

[19]  N. Busis,et al.  Modulation of synapse formation by cyclic adenosine monophosphate. , 1983, Science.

[20]  D. Jacobowitz,et al.  A primate model of parkinsonism: selective destruction of dopaminergic neurons in the pars compacta of the substantia nigra by N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[21]  C. Johnson,et al.  A single-vial biphasic liquid extraction assay for choline acetyltransferase using [3H]choline. , 1981, Analytical biochemistry.

[22]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[23]  W. Shain,et al.  Neuronal properties of hybrid neuroblastoma X sympathetic ganglion cells. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[24]  J. Minna,et al.  Expression of phenotypes in hybrid somatic cells derived from the nervous system. , 1975, Genetics.

[25]  K. Jellinger,et al.  Brain dopamine and the syndromes of Parkinson and Huntington. Clinical, morphological and neurochemical correlations. , 1973, Journal of the neurological sciences.

[26]  R. Johnson,et al.  Premature chromosome condensation: a mechanism for the elimination of chromosomes in virus-fused cells. , 1972, Journal of cell science.

[27]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[28]  M. Nirenberg,et al.  Neuronal tumor cells with excitable membranes grown in vitro. , 1969, Proceedings of the National Academy of Sciences of the United States of America.

[29]  B. Ephrussi,et al.  Studies of Interspecific (Rat x Mouse) Somatic Hybrids. II. Lactate Dehydrogenase and beta-GLUCURONIDASE. , 1966, Genetics.

[30]  J. Littlefield Selection of Hybrids from Matings of Fibroblasts in vitro and Their Presumed Recombinants , 1964, Science.

[31]  Body and brain. , 1994, Journal of the Royal Society of Medicine.

[32]  J. Bostwick,et al.  A tyrosine hydroxylase assay in microwells using coupled nonenzymatic decarboxylation of dopa. , 1991, Analytical biochemistry.

[33]  R. Oppenheim Cell death during development of the nervous system. , 1991, Annual review of neuroscience.

[34]  A. Dahlström,et al.  Investigations on auto-antibodies in Alzheimer's and Parkinson's diseases, using defined neuronal cultures. , 1990, Journal of neural transmission. Supplementum.

[35]  M. Tohyama,et al.  Distribution of the omega-conotoxin receptor in rat brain. An autoradiographic mapping. , 1989, Neuroscience.

[36]  H. Glossmann,et al.  Assay for calcium channels. , 1985, Methods in enzymology.

[37]  J. Penney,et al.  Speculations on the functional anatomy of basal ganglia disorders. , 1983, Annual review of neuroscience.

[38]  R. Rips,et al.  A New Technique for Simultaneous Assay of Biogenic Amines and Their Metabolites in Unpurified Mouse Brain , 1982 .

[39]  G H Sato,et al.  Growth of a rat neuroblastoma cell line in serum-free supplemented medium. , 1979, Proceedings of the National Academy of Sciences of the United States of America.