Genomic organisation and functional expression of the gene encoding the human serotonin 5-HT2C receptor.
The 5-HT2C receptor gene is unique among the members of the 5-HT receptor family by virtue of its genomic organisation. The human 5-HT2C receptor gene, unlike many other genes for guanine nucleotide binding (G)-proteins, contains three introns which interrupt the coding sequence into four exons. The first two introns are at equivalent positions as compared to the intervening sequences previously found in the 5-HT2(A) receptor gene, suggesting a close evolutionary relationship between both genes. Southern blot analysis shows that the 5-HT2C receptor gene is a single copy gene. Furthermore, we report the functional expression of a complementary DNA for the 5-HT2C receptor, cloned from hippocampal RNA. Membranes prepared from NIH 3T3 cells stably expressing the 5-HT2C receptor cDNA, displayed a single population of high affinity sites for the antagonist [3H]mesulergine (Kd = 2.9 +/- 0.4 nM, Bmax = 44.3 +/- 7.2 pmol/mg protein) as well as for [3H]5-HT (Kd = 9.9 +/- 0.7 nM, Bmax = 13.6 +/- 1.0 pmol/mg protein). Displacement of [3H]mesulergine and [3H]5HT binding by ligands indicated a pharmacological similarity of these binding sites with porcine and rat choroid plexus 5-HT2C receptors. Furthermore, activation of the 5-HT2C receptor with 5-HT results in an increased phospholipase C activity.
Stam
N J
NJ
Department of Biotechnology and Biochemistry, N.V. Organon.
Vanderheyden
P
P
van Alebeek
C
C
Klomp
J
J
de Boer
T
T
van Delft
A M
AM
Olijve
W
W
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D000978:Antiparkinson Agents / Q000494:pharmacology; D001483:Base Sequence; D001667:Binding, Competitive; D015139:Blotting, Southern; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004873:Ergolines / Q000378:metabolism / Q000494:pharmacology; D005091:Exons; D005786:Gene Expression Regulation / Q000235:genetics*; D006624:Hippocampus / Q000378:metabolism; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D044405:Receptor, Serotonin, 5-HT2C; D011985:Receptors, Serotonin / Q000737:chemistry / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D015183:Restriction Mapping; D014162:Transfection; D010738:Type C Phospholipases / Q000378:metabolism
D000978:Antiparkinson Agents; D017931:DNA Primers; D018076:DNA, Complementary; D004873:Ergolines; D012333:RNA, Messenger; D044405:Receptor, Serotonin, 5-HT2C; D011985:Receptors, Serotonin; D010738:Type C Phospholipases; C031649:mesulergine