The cloning and chromosomal mapping of two novel human opioid-somatostatin-like receptor genes, GPR7 and GPR8, expressed in discrete areas of the brain.
Following the cloning of the opioid receptors mu, kappa, and delta, we conducted a search for related receptors. Using oligonucleotides based on the opioid and also the structurally related somatostatin receptors, we amplified genomic DNA using the polymerase chain reaction and isolated fragments of novel G protein-coupled receptor genes. Two of these gene fragments designated clones 12 and 11 were used to isolate the full-length genes. The intronless coding sequences of these genes, named GPR7 and GPR8, shared 70% identity with each other, and each shared significant similarity with the sequences encoding transmembrane regions of the opioid and somatostatin receptors. GPR7 was mapped to chromosome 10q11.2-q21.1 and GPR8 to chromosome 20q13.3. Northern blot analysis using human mRNA demonstrated expression of GPR7 mainly in cerebellum and frontal cortex, while GPR8 was located mainly in the frontal cortex. In situ hybridization revealed expression of GPR7 in the human pituitary. A partial sequence of the mouse orthologue of GPR7 was obtained, and in situ hybridization demonstrated expression in discrete nuclei of brain, namely suprachiasmatic, arcuate, and ventromedial nuclei of hypothalamus. A stable cell line expressing the GPR7 gene was created, but expression levels of the receptor were low. The available pharmacology indicated binding to several opioid drugs such as bremazocine, levorphanol, and beta-FNA, but not to the opioid receptor subtype-selective mu, delta, or kappa agonists.
O'Dowd
B F
BF
Addiction Research Foundation, Toronto, Ontario, Canada.
Scheideler
M A
MA
Nguyen
T
T
Cheng
R
R
Rasmussen
J S
JS
Marchese
A
A
Zastawny
R
R
Heng
H H
HH
Tsui
L C
LC
Shi
X
X
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001575:Benzomorphans / Q000378:metabolism; D001921:Brain / Q000378:metabolism*; D002874:Chromosome Mapping; D002879:Chromosomes, Human, Pair 10; D002883:Chromosomes, Human, Pair 14; D002890:Chromosomes, Human, Pair 20; D003001:Cloning, Molecular; D006801:Humans; D017403:In Situ Hybridization; D008024:Ligands; D051379:Mice; D008969:Molecular Sequence Data; D011869:Radioligand Assay; D011957:Receptors, Opioid / Q000235:genetics* / Q000378:metabolism; D017481:Receptors, Somatostatin / Q000235:genetics* / Q000378:metabolism
D001575:Benzomorphans; D008024:Ligands; D011957:Receptors, Opioid; D017481:Receptors, Somatostatin; C027248:bremazocine