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Cloning of a C-terminally truncated NK-1 receptor from guinea-pig nervous system.

Author	Sarah J Baker, Judy L Morris, Ian L Gibbins, Sarah J Baker, Judy L Morris, Ian L Gibbins
Citation Information	Brain research. Molecular brain research, 111:136-47 (2003)
Keywords	Alternative Splicing, Amino Acid Sequence, Animals, Base Sequence, Central Nervous System, Cloning, Molecular, DNA, Complementary, Dendrites, Female, Fluorescent Antibody Technique, Ganglia, Sympathetic, Guinea Pigs, Male, Microscopy, Confocal, Microtubule-Associated Proteins, Molecular Sequence Data, Mutation, Neurons, Peripheral Nervous System, Protein Isoforms, Protein Structure, Tertiary, RNA, Messenger, Receptors, Neurokinin-1
Applications	Immunofluorescence
Species:	Guinea Pig
Related Products	AB5060
Pub Med ID	12654513

In order to examine the possibility that some actions of substance P may be mediated by a variant of the neurokinin-1 (NK-1) receptor, we isolated and sequenced the cDNA encoding a truncated NK-1 receptor from guinea-pig celiac ganglion and brain mRNA by two-step RT-PCR based on the 3'RACE method. The truncated NK-1 receptor sequence corresponded to a splice variant missing the final exon 5, and encoded a 311-amino acid protein that was truncated just after transmembrane domain 7, in an identical position to a truncated variant of the human NK-1 receptor. Thus, the truncated NK-1 receptor lacked the intracellular C-terminus sequence required for the phosphorylation and internalisation of the full-length NK-1 receptor. Using a sensitive one-step semi-quantitative RT-PCR assay, we detected mRNA for both the full length and truncated NK-1 receptors throughout the brain, spinal cord, sensory and autonomic ganglia, and viscera. Truncated NK-1 receptor mRNA was present in lower quantities than mRNA for the full-length NK-1R in all tissues. Highest levels of mRNA for the truncated NK-1 receptor were detected in coeliac ganglion, spinal cord, basal ganglia and hypothalamus. An antiserum to the N-terminus of the NK-1 receptor labelled dendrites of coeliac ganglion neurons that were not labelled with antisera to the C-terminus of the full length NK-1 receptor. These results show that a C-terminally truncated variant of the NK-1 receptor is likely to be widespread in central and peripheral nervous tissue. We predict that this receptor will mediate actions of substance P on neurons where immunohistochemical evidence for a full-length NK-1 receptor is lacking.