Characterization, Neurosteroid Binding and Brain Distribution of Human Membrane Progesterone Receptors (cid:1) and (cid:2) (mPR (cid:1) and mPR (cid:2) ) and mPR (cid:1) Involvement in Neurosteroid Inhibition of Apoptosis

ThreemembersoftheprogestinandadipoQreceptor(PAQR)family,PAQR-7,PAQR-8,andPAQR-5 [membrane progesterone (P4) receptor (PR) (mPR) (cid:1) , mPR (cid:2) , and mPR (cid:3) ], function as plasma mPRs coupledtoGproteinsinmammaliancells,butthecharacteristicsoftwoothermembers,PAQR6and PAQR9 (mPR (cid:4) and mPR (cid:5) ), remain unclear, because they have only been investigated in yeast expression systems. Here, we show that recombinant human mPR (cid:4) and mPR (cid:5) expressed in MDA-MB-231 breast cancer cells display specific, saturable, high-affinity [ 3 H]-P4 binding on the plasma membranes of transfected cells with equilibrium dissociation constants (K d s) of 2.71 and 2.85 n M , respectively, and low affinity for R5020, characteristics typical of mPRs. P4 treatment increased cAMP production as well as [ 35 S]-guanosine 5 = -triphosphate (GTP) (cid:3) S binding to transfected cell membranes, which was immunoprecipitated with a stimulatory G protein antibody, suggesting both mPR (cid:4) and mPR (cid:5) activate a stimulatory G protein (Gs), unlike other mPRs, which activate an inhibitory G protein (Gi). All five mPR mRNAs were detected in different regions of the human brain, but mPR (cid:4) showed greatest expression in many regions, including the forebrain, hypothalamus, amygdala, corpus callosum, and spinal cord, whereas mPR (cid:5) was abundant in the pituitary gland and hypothalamus. Allopregnanolone and other neurosteroids bound to mPR (cid:4) and other mPRs and acted as agonists, activating second messengers and decreased starvation-induced cell deathandapoptosisinmPR (cid:4) -transfectedcellsandinhippocampalneuronalcellsatlownanomolar concentrations. The results suggest that mPR (cid:4) and mPR (cid:5) function as mPRs coupled to G proteins and are potential intermediaries of nonclassical antiapoptotic actions of neurosteroids in the central nervous system. ( Endocrinology 154: 283–295, 2013)

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