Radioimmunoassay of rat prolactin and its use in measuring prolactin production by cultured pituitary cells

Abstract

A sensitive and specific radioimmunoassay has been developed for rat prolactin (rPRL), employing the double antibody solid phase technique for the separation of free and antibody-bound [125I]rPRL. The anti-serum was raised in rabbits and showed no cross-reaction with rat growth hormone (rGH), follicle stimulating hormone (rFSH), luteinizing hormone (rLH) and thyrotrophin (rTSH). The immunosorbent (sheep anti-rabbit IgG bound to cellulose) showed a surprisingly high binding of [125I]rPRL, but not of the other iodinated anterior pituitary hormones. Addition of serum to the incubation mixtures prevented the binding between [125I]rPRL and the immunosorbent. Three different clonal strains of pituitary cells have been examined for production of rPRL, rLH and rTSH, both in the basal state as well as after treatment with thyrotrophin releasing hormone (TRH) and gonadotrophin releasing hormone (LH/FSH-RH). Monolayer cultures of two of the cell strains produced and secreted rPRL spontaneously, and they showed a 2-fold increase in rPRL production after treatment with TRH (3-10(-7) mol/1). The third cell strain did not produce rPRL spontaneously, or after treatment with TRH. None of these cell strains could be stimulated to produce rTSH by treatment with TRH. Treatment of the same three cell strains with LH/FSH-RH (1.2-10(-6) mol/1) failed to induce production of rLH, and there were no changes in production of rPRL. Prostaglandins E1 and E2 (3-10(-8) mol/1) and oestradiol-17beta (10(-7)-10(-10) mol/1), however, stimulated the production of rPRL. The effects of TRH and prostaglandins E1 and E2 were observed within 24 h of treatment, while the first effect of oestradiol-17beta was seen after 3 days. These results suggest that the stimulatory effect of oestradiol-17beta on rPRL production differs from that of TRH and prostaglandins.