Gene and cDNA cloning and characterization of the mouse V3/V1b pituitary vasopressin receptor.
The gene of the mouse V3/V1b receptor was identified by homology cloning. One of the genomic clones contained the entire coding sequence. The cDNA presented high identity with rat (92%) and human (84%) sequences. Southern blot analysis indicated the existence of a single gene. Tissue distribution was studied by RT-PCR. The major site of expression was the pituitary. A faint signal was also present in hypothalamus, brain, adrenal, pancreas and colon. The mouse corticotroph cell line, AtT20, did not express the transcript. In order to confirm the identity of the sequence, the V3/V1b receptor cDNA was cloned and stably expressed in CHO-AA8 Tet-Off cells under the control of tetracycline. When transfected cells were treated with arginine vasopressin (AVP), inositol phosphate production increased in a dose-dependent manner, indicating that the V3/V1b receptor couples to phospholipase C. Moreover, AVP did not stimulate cAMP production. Binding studies with [3H]AVP indicated that the affinity of the mouse V3/V1b receptor (Kd=0.5 nM) is similar to that reported for rat and human receptors. The rank order of potency established in competition binding experiments with different analogues was representative of a V3/V1b profile, distinct from V1a and V2. However, significant differences were found between human and mouse receptors tested in parallel. Thus the pharmacology of V3/V1b receptors can not be transposed among different species.
Ventura
M A
MA
Groupe d'Etudes en Physiopathologie Endocrinienne, UPR 1524 CNRS, Institut Cochin de Génétique Moléculaire, CHU Cochin, Université Paris V, 75014 Paris.
René
P
P
de Keyzer
Y
Y
Bertagna
X
X
Clauser
E
E
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D019556:COS Cells; D003001:Cloning, Molecular; D003062:Codon; D006224:Cricetinae; D018076:DNA, Complementary; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010902:Pituitary Gland / Q000378:metabolism*; D014176:Protein Biosynthesis; D011869:Radioligand Assay; D017483:Receptors, Vasopressin / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D017386:Sequence Homology, Amino Acid
D003062:Codon; D018076:DNA, Complementary; D017483:Receptors, Vasopressin