Identification of 5-HT receptor sub-types in a homogeneous population of presumptive serotoninergic neurones.
The expression of the known rat 5-HT receptor sub-types has been analyzed in a presumptive serotoninergic cell line derived from the rat raphé nuclei, using reverse transcription followed by polymerase chain reaction. By manipulating the activity of the oncogene (ts-SV40T) product used to immortalize the serotoninergic precursors, it has been possible to compare the expression of the 5-HT receptors in either replicative or differentiating cells. 5-HT1B, 5-HT1D, 5-HT3, 5-HT6 and 5-HT7 receptor gene expression were all observed in the replicating cells. However, under differentiation conditions, expression of all except the 5-HT1B receptor was lost. Only one novel amplification product appeared during early differentiation, in the 5-HT2B lane; its smaller than expected size was suggestive of a previously undescribed alternate splicing of the mRNA in brain. The curtailment of 5-HT receptor expression in differentiating neurones in vitro may reflect the normal ongoing restriction in the phenotypic potential during embryogenesis in vivo. The serotonin cells, therefore, constitute a pristine cell line in which to study the receptor pharmacology of one or more 5-HT receptor sub-types in isolation.
Jackson
Z E
ZE
School of Molecular and Medical Biosciences, University of Wales, Cardiff, U.K.
Stringer
B M
BM
Foster
G A
GA
D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D004587:Electrophoresis, Agar Gel; D008526:Medulla Oblongata / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008636:Mesencephalon / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008969:Molecular Sequence Data; D009474:Neurons / Q000378:metabolism*; D015345:Oligonucleotide Probes; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D051381:Rats; D011985:Receptors, Serotonin / Q000096:biosynthesis* / Q000235:genetics; D012701:Serotonin / Q000502:physiology*
D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D011985:Receptors, Serotonin; D012701:Serotonin