High tumour prolactin receptor content and lack of increase in serum prolactin levels as predictors of good response to endocrine therapy in rat mammary cancer.

Correlations between anti-neoplastic activity of medroxyprogesterone acetate (MPA), on the one hand, and serum prolactin (PRL) levels as well as tumour PRL and insulin receptor content, on the other, were investigated in female rats bearing dimethylbenzanthracene (DMBA)-induced mammary tumours. Changes in liver PRL receptor concentrations were also studied. MPA was injected for 15 days. Regression was observed in 16 out of 50 (32%) tumours from rats treated with MPA. Twenty-seven out of 50 (54%) continued to grow regardless of treatment. Stasis was seen in the remaining 7 tumours (14%). Serum PRL levels increased significantly in rats with tumours which were non-responsive to MPA. Concentration of PRL receptors in the liver of all animals was reduced by MPA treatment. A remarkable increase occurred only in those mammary tumours which responded to therapy. The concentrations of PRL receptors in the tumours non-responsive to MPA were similar to those detected in control tumours. Unlike PRL receptors, tumour insulin receptor levels were not modified by MPA treatment. Five out of 14 tumours (35.7%), previously growing in spite of MPA administration, regressed when bromocriptine was added to MPA. A significant reduction in serum PRL levels occurred in all rats undergoing the latter treatment. No difference was observed between responsive and non-responsive animals; on the contrary, the PRL receptor content of responsive tumours increased significantly in comparison with that of non-responsive tumours.

Prolactin receptors on large granular lymphocytes: dual regulation by cyclosporin A.

Although evidence has been provided for a modulatory role of prolactin (PRL) on humoral and cell-mediated immune responses and PRL receptors have been found on T and B lymphocytes, no indications exist concerning the influence of PRL on natural killer (NK) activity nor has a structural basis for interaction been found on the NK effector cells (large granular lymphocytes, LGL). We show here that highly purified LGL express binding sites for PRL. The calculated receptor number was 660 per cell and the dissociation constant (Kd) was 3.0 X 10(-10) M. Since previous studies have reported that cyclosporin (CsA), an immunosuppressive agent used in organ transplant patients, affects the binding of PRL to T and B lymphocytes, but not to rabbit mammary gland cells, we investigated whether this compound could alter the binding of the hormone to LGL. At concentrations from 10(-7) to 10(-6), corresponding to the therapeutical range, CsA induced a complete inhibition of the PRL binding. By contrast, concentrations of CsA ranging from 10(-11) to 10(-9) increased the PRL binding to more than 100% of control levels. In addition to their antitumor role, LGL have been proposed to participate in graft versus host disease and in transplant rejection. The finding that CsA can differently affect PRL-receptor expression on LGL points to an involvement of CsA--PRL interactions in determining the output of these immune responses. In addition, these data strongly support the idea of a close relationship between the neuroendocrine and immune systems.

Regional distribution and species variation of prolactin binding sites in the brain.

The binding of 125I-labeled ovine prolactin (125I-oPRL) to membranes from different brain regions of pigeon, rabbit, rat, pig, calf, horse, and ewe was studied. The hypothalamus from rabbit, pig, horse, and pigeon showed a low but specific binding for 125I-oPRL clearly different from the other brain regions examined (cortex and cerebellum), whereas in the brain from rat, calf, and ewe the binding was very small and quite uniform in the various regions. Also the membranes from choroid plexus of rabbit, pig, calf, and horse showed an evident specific binding for prolactin. The binding of 125I-oPRL to hypothalamus and choroid plexus membranes from rabbit and horse was inhibited in a dose-dependent manner by unlabeled oPRL and hGH but not by many other polypeptide hormones. Scatchard analysis of the binding revealed the presence of binding sites with low capacity and high affinity for ovine ligand.

Phosphatidylserine activity on prolactin brain receptors of aged rabbits.

1. PRL receptors in the hypothalamus and substantia nigra of aged rabbits (28-month-old) are significantly reduced in comparison with young rabbits (6-month-old). 2. Repeated treatments with BC-PS are able to gradually increase the PRL receptor number both in hypothalamus and nigra. However only after 30 days of treatment the binding reaches the mean values observed in young rabbits. 3. Aged rabbits showed an evident increase in PRL plasma levels in comparison with young animals. In BC-PS treated animals this increase was not more apparent. Moreover in young rabbits treated with BC-PS an evident decrease in basal PRL plasma levels was observed.

Prolactin binding sites in human erythrocytes and lymphocytes.

Specific binding sites for prolactin (PRL) have been studied in human peripheral lymphocytes and erythrocytes of normal adult volunteers and of term cord bloods. In erythrocytes from healthy adult subjects of both sexes a very low specific binding of 125I-human PRL was found (0.24%), whereas a higher binding was found in term cord blood (1.1%). The binding was hormone specific, the binding capacity was 2.6 fmol/4 X 10(9) cells and the Kd was 3.4 X 10(-10) M. In lymphocytes of both adults and term cord bloods an evident specific binding was observed (male adults: 1.6%; female adults: 1.7%; cord blood: 1.8%). The binding was specific for lactogenic hormones and the binding capacity was 3.7 fmol/2 X 10(6) cells and the Kd was 3.9 X 10(-10) M. The presence of specific binding sites for PRL on human erythrocytes and lymphocytes could be used to study PRL binding on blood cells of patients in different physiological or pathological situations.

Prolactin binding sites in Xenopus laevis tissues: comparison between normal and dehydrated animals.

The binding of 125I-labeled ovine prolactin (125I-oPRL) to membranes from the kidney, epidermis, liver, and testis of Xenopus laevis adult specimens either kept in an aquatic environment or exposed for 2 weeks to dehydrating conditions was studied. Prolactin binding specificity was assayed through competition with several unlabeled hormones (oPRL, hGH, rGH, rLH, and porcine insulin). In the animal exposed to dehydrating conditions a statistically highly significant reduction in prolactin binding to the membranes from the kidney and epidermis was recorded. No significant variations were revealed by the membranes from the liver and testis. The reduction detected in the binding of 125I-oPRL is not related to the dissociation constant, but to the number of PRL binding sites. Since PRL ranks among the few peptide hormones whose rise in the bloodstream promotes an increase in the number of their own receptors, the reduction of its binding sites in Xenopus specimens exposed to dehydration might lend some support to our earlier hypothesis that transfer to a dehydrating environment may bring about, in this totally aquatic species, some decrease in the blood PRL levels.

Further evidence for the presence of specific binding sites for prolactin in the rabbit brain. Preferential distribution in the hypothalamus and substantia nigra.

In the present research we have extended our work on the presence of binding sites for prolactin in the rabbit brain focusing our attention on the brain areas with high dopamine cell bodies density. Among these areas the hypothalamus showed the highest specific binding of labeled ovine prolactin (oPRL). Clearly detectible specific binding was observed also in substantia nigra, whereas in other brain regions the specific binding was very small, except for the striatum where a low but not negligible binding was found in female rabbits. The binding of 125I-oPRL showed a hormonal specificity and Scatchard analysis of the binding showed no clear difference in dissociation constant (Kd) between hypothalamus, nigra and striatum.

Presence and characterization of prolactin receptors in human benign breast tumours.

Prolactin receptors have been determined in 64 benign breast tumours. A specific binding of 0.5% or more (with a range of 0.5-3.3%) was found in 34.4% of the cases and was considered prolactin-receptor-positive. The binding was found to be specific only for lactogenic hormones. By Scatchard analysis the dissociation constant was 2.55 X 10(-10) mol/l and the binding capacity was 4.6 fmol/mg protein.

Modifications in prolactin binding capacity in the rat liver induced by non-steroidal anti-inflammatory drugs.

Modifications in prolactin specific binding in the rat liver induced by different non-steroidal anti-inflammatory drugs (indomethacin, piroxicam, ketoprofen, phenylbutazone, mefenamic acid and acetylsalicylic acid) have been studied. All caused a dose-dependent inhibition of prolactin binding capacity whereas no change was found in dissociation constant values. The inhibitory effect is reversible and highly specific, since insulin binding to the same membrane preparation is not affected. The degree of inhibitory activity on prolactin binding is related to the anti-inflammatory activity of each drug, which supports the hypothesis of an involvement of prostaglandin synthesis.

Physiological and pharmacological variations in rabbit prolactin plasma levels.

By using a specific homologous double-antibody RIA, physiological and pharmacological variation in prolactin plasma levels were studied in the rabbit. The study of plasma levels during 24 h period demonstrated the presence of a rhythmic secretion of prolactin with higher values between 15.00 and 19.00 h. Prolactin plasma levels were low in neonatal rabbits and increased gradually with the age of the animals. In adult rabbits a significant higher prolactin plasma concentration was found in female animals. Blood levels fluctuate during the first half of pregnancy but the mean levels were higher than those found during the second half of gestation. A remarkable increase of plasma levels was observed 24 h before parturition and during lactation. Plasma prolactin concentrations increased after injection of both chlorpromazine and sulpiride. The hyperprolactinaemic effect of sulpiride was abolished by bromocriptine.