Cloning of a receptor for prostaglandin F2 alpha from the ovine corpus luteum.
A complementary DNA clone encoding a functional receptor for prostaglandin F2 alpha (PGF2 alpha) has been isolated from an ovine large luteal cell complementary DNA library (prepared from day 10 mid-luteal phase RNA). This receptor, which has been designated FP, consists of 362 amino acids (M(r) = 40,982) and is a member of the family of G protein-coupled receptors. Radioligand binding studies with membranes prepared from transfected COS cells demonstrated specific 17-[3H]phenyl-trinor-PGF2 alpha binding that was displaced by prostanoids in the order of 17-phenyl-trinor-PGF2 alpha > PGF2 alpha > fluprostenol > PGD2 > PGE2 >> 8-epi PGF2 alpha. Xenopus laevis oocytes injected with RNA encoding the ovine FP receptor responded to 17-phenyl-trinor-PGF2 alpha with increased membrane chloride conductance in calcium-free medium. Northern blot analysis with RNA from day 10 corpus luteum showed a major band of approximately 6.1 kilobases. On day 14, when luteolysis usually starts, the abundance of this 6.1-kilobase band was variable between individual ewes, and on day 16, when luteolysis is underway, the message was uniformly less abundant. This variability appeared to correlate with circulating progesterone. Thus, when the progesterone level was high (days 10 and 14 depending on whether luteolysis had started), the amount of FP receptor message was high, whereas when the progesterone level was low or falling (day 16), the amount of FP receptor message decreased. We have cloned an ovine FP receptor whose expression confers appropriate functional activity in COS cells and Xenopus oocytes. Furthermore, the level of messenger RNA encoding the FP receptor is high in the midluteal phase ovine corpus luteum and decreases during luteolysis.
Graves
P E
PE
Department of Pharmacology and Toxicology, University of Arizona, Tucson 85721, USA.
Pierce
K L
KL
Bailey
T J
TJ
Rueda
B R
BR
Gil
D W
DW
Woodward
D F
DF
Yool
A J
AJ
Hoyer
P B
PB
Regan
J W
JW
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D003338:Corpus Luteum / Q000502:physiology*; D005260:Female; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism; D011982:Receptors, Prostaglandin / Q000235:genetics* / Q000378:metabolism; D012756:Sheep / Q000235:genetics*; D014982:Xenopus laevis
D011982:Receptors, Prostaglandin; C034671:prostaglandin F2alpha receptor