1. Cloning and characterization of a novel human 5-HT4 receptor variant that lacks the alternatively spliced carboxy terminal exon. RT-PCR distribution in human brain and periphery of multiple 5-HT4 receptor variants
- Author
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T Doménech, José Palacios, Guadalupe Mengod, and M T Vilaró
- Subjects
Male ,Agonist ,DNA, Complementary ,medicine.drug_class ,In Vitro Techniques ,Biology ,[3H]GR 113808 ,5-HT4 receptors ,Hippocampus ,Cell Line ,5-HT7 receptor ,Radioligand Assay ,Cellular and Molecular Neuroscience ,Exon ,Receptor constitutive activity ,Peripheral Nervous System ,Cyclic AMP ,medicine ,Animals ,Humans ,splice ,RNA, Messenger ,Cloning, Molecular ,Receptor ,Brain Chemistry ,Pharmacology ,Genetics ,Reverse Transcriptase Polymerase Chain Reaction ,Cell Membrane ,Alternative splicing ,Exons ,Oligonucleotides, Antisense ,musculoskeletal system ,Receptor antagonist ,Molecular biology ,Alternative Splicing ,Macaca fascicularis ,C-terminal splice variants ,Receptors, Serotonin ,RNA splicing ,Human brain distribution ,Female ,Receptors, Serotonin, 5-HT4 - Abstract
We have cloned a novel C-terminal splice variant of serotonin 5-HT4 receptors from human hippocampus. The deduced protein extends only one aminoacid past the splicing point. We propose to call the novel variant h5-HT4(n) since it contains none of the C-terminal exons alternatively spliced in other variants. The pharmacological profile of h5-HT4(n) stably expressed in HeLa cells is in agreement with other reported variants. Stably transfected cells showed increased basal levels of intracellular cAMP in absence of agonist, indicating constitutive activity of the expressed receptors. 5-HT induced robust increases of intracellular cAMP. The 5-HT4 receptor antagonist GR 113808 blocked the effects of 5-HT and brought intracellular cAMP below basal constitutive levels, indicating inverse agonism of this compound in this system. The RT-PCR distribution of all known human C-terminal splice variants in human brain regions and periphery showed complex patterns of variant expression, with the novel variant h5-HT4(n) being widely and abundantly expressed. © 2002 Elsevier Science Ltd. All rights reserved., This work was supported by grants from CICYT, SAF 96-0336 and SAF 97-0117. In the very initial phases of this project, M.T.V. was recipient of a Long Term EMBO fellowship (ALTF 448-1992). Support from the CIRIT (Generalitat de Catalunya) to the Department of Neurochemistry (IIBB/CSIC) as Grup de Recerca Consolidat (1999 SGR-00210) is acknowledged
- Published
- 2002