Molecular cloning and characterization of the mouse P2Y(4) nucleotide receptor

To isolate the mouse P2Y(4) receptor gene, a mouse genomic Library was scre ened with a human P2Y(4) probe. An open reading frame encoding a protein of 361 amino acids was isolated. This protein showed 82% and 95% amino acid i dentity with the human and rat P2Y(4) receptors, respectively. By reverse t ranscription and polymerase chain reaction (RT-PCR), the P2Y(4) messenger R NA was detected in mouse Liver, intestine, stomach, bladder and lung among the 16 mouse tissues tested. In 1321N1 transfected cells, the mouse P2Y(4) receptor was equally activated by UTP and ATP, and was antagonized by pyrid oxal-phosphate-6-azophenyl-2 ' ,4 ' -disulphonic acid (PPADS) and Reactive Blue 2, and not by suramin. Moreover, when expressed in 1321N1 cells, the r at P2Y(4) is also antagonized by PPADS. Thus, when compared in the same exp ression system, the mouse P2Y(4) is closer to the rat ortholog in terms of a,agonist stimulation, while in terms of antagonist profile, the three P2Y( 4) receptor orthologs are similar. (C) 2001 Elsevier Science B.V. All right s reserved.