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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1 Pt 2
pubmed:dateCreated
1995-2-16
pubmed:abstractText
Immortalized hybrid cells were generated by somatic cell fusion of 18-d-old embryonic corpus striatum of the mouse strain C57BL/6J with the N18TG2 neuroblastoma. One of the cell populations obtained was treated with a combination of 1 mM n-butyric acid and 10 microM SKF 38393 (a specific D1 agonist), and a surviving cell population (E1X) was subcloned. Twenty-seven monoclonal cell lines were obtained and screened for the expression of striatal-specific characteristics including gamma-aminobutyric acid (GABA), choline acetyltransferase (ChAT), acetylcholine (ACh), mRNA for specific dopamine receptors, and dopamine- and adenosine 3':5'-monophosphate-regulated phosphoprotein, M(r) 32,000 (DARPP-32), and functional D1 and D2 dopamine receptors. Neither the parent hybrid cell population (E1X) nor any of the monoclonal cell lines examined expressed GABA levels significantly different than that of the N18TG2 parent neuroblastoma cells (1.36 +/- 0.07 micrograms/mg protein). The range of ChAT activity in the monoclonal hybrid cell lines was 5.5 +/- 0.3 to 921.3 +/- 97.4 pmol/min/mg protein. Two of the cell lines expressing ChAT activity (X52 and X58) contained ACh (49.64 +/- 4.23 and 1.78 +/- 0.07 ng/mg protein, respectively). The neuronal origin of four of the monoclonal hybrid lines was shown by their immunoreactivity, following differentiation with 10 microM forskolin, to neurofilament protein, a neuron-specific marker. The monoclonal hybrid cell lines, but not the N18TG2 neuroblastoma, were shown to express an array of D1, D2, and D5 receptor mRNA as well as DARPP-32 mRNA. Two monoclonal cell lines expressed D1 receptor binding sites (X57, 29.2 +/- 4.5 fmol/mg protein and X62, 43.8 +/- 6.8 fmol/mg protein) which mediated the stimulation of adenylate cyclase activity. One cell line, X58, expressed only D2 dopamine receptors (80.9 +/- 9.8 fmol/mg protein) which were negatively coupled to adenylate cyclase activity. These findings suggest that the immortalized monoclonal hybrid cell lines are of neuronal origin and have incorporated elements of the medium spiny and cholinergic neurons of the developing striatum.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jan
pubmed:issn
0270-6474
pubmed:author
pubmed:issnType
Print
pubmed:volume
15
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
676-88
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:7823171-Acetylcholine, pubmed-meshheading:7823171-Acetylcholinesterase, pubmed-meshheading:7823171-Adenylate Cyclase, pubmed-meshheading:7823171-Animals, pubmed-meshheading:7823171-Base Sequence, pubmed-meshheading:7823171-Binding Sites, pubmed-meshheading:7823171-Biological Markers, pubmed-meshheading:7823171-Cell Line, pubmed-meshheading:7823171-Corpus Striatum, pubmed-meshheading:7823171-Cytoskeleton, pubmed-meshheading:7823171-Dopamine and cAMP-Regulated Phosphoprotein 32, pubmed-meshheading:7823171-Hybrid Cells, pubmed-meshheading:7823171-Mice, pubmed-meshheading:7823171-Mice, Inbred C57BL, pubmed-meshheading:7823171-Molecular Sequence Data, pubmed-meshheading:7823171-Nerve Tissue Proteins, pubmed-meshheading:7823171-Phosphoproteins, pubmed-meshheading:7823171-RNA, Messenger, pubmed-meshheading:7823171-Receptors, Dopamine
pubmed:year
1995
pubmed:articleTitle
Immortalized murine striatal neuronal cell lines expressing dopamine receptors and cholinergic properties.
pubmed:affiliation
Department of Pharmacological and Physiological Sciences, University of Chicago, Illinois 60637.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't