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MODEL ORGANISMS AND COMPARATIVE FUNCTIONAL GENOMICS

Molecular cloning, characterization, and distribution of the gerbil angiotensin II AT₂ receptor

¹Section on Pharmacology, National Institute of Mental Health, ²Gene Targeting Facility, National Institute of Dental and Craniofacial Research, and ³Cell and Cancer Biology Branch, National Cancer Institute, National Institutes of Health, Department of Health and Human Services, Bethesda, Maryland 20892-1514

Submitted 7 January 2003 ; accepted in final form 4 August 2003

We isolated a cDNA clone encoding the gerbil AT₂ receptor (gAT₂) gene from a gerbil adrenal gland cDNA library. The full-length cDNA contains a 1,089-bp open reading frame encoding 363 amino acid residues with 90.9, 96.1, and 95.6% identity with the human (hAT₂), rat (rAT₂), and mouse AT₂ (mAT₂) receptors, respectively. There are at least seven nonconserved amino acids in the NH₂-terminal domain and in positions Val¹⁹⁶, Val²¹⁷, and Met²⁹³, important for angiotensin (ANG) II but not for CGP-42112 binding. Displacement studies in adrenal sections revealed that affinity of the gAT₂ receptor was 10-20 times lower for ANG II, ANG III, and PD-123319 than was affinity of the rAT₂ receptor. The affinity of each receptor remained the same for CGP-42112. When transfected into COS-7 cells, the gAT₂ receptor shows affinity for ANG II that is three times lower than that shown by the hAT₂ receptor, whereas affinities for ANG III and the AT₂ receptor ligands CGP-42112 and PD-123319 were similar. Autoradiography in sections of the gerbil head showed higher binding in muscles, retina, skin, and molars at embryonic day 19 than at 1 wk of age. In situ hybridization and emulsion autoradiography revealed that at embryonic day 19 the gAT₂ receptor mRNA was highly localized to the base of the dental papilla of maxillary and mandibular molars. Our results suggest selective growth-related functions in late gestation and early postnatal periods for the gAT₂ receptor and provide an essential basis for future mutagenesis studies to further define structural requirements for agonist binding.

adrenal gland; brain; CGP-42112; dental papilla