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The isolation and identification of rat prolactin (Kwa, 1965; Kwa, van der Bent & Prop, 1967) made it possible to develop a radioimmunoassay method for the estimation of prolactin in plasma and in pituitary tissue (Kwa & Verhofstad, 1967). Results are now given for plasma levels of prolactin, expressed as ng./ml. in rats of the highly inbred strains U and R, Amsterdam and the F 1 hybrids (R x U) during their oestrous cycle, using the rat prolactin prepared by the above mentioned method as standard (15·6 i.u./mg.).
Daily vaginal smears were taken from the age of 10 to 12 weeks from rats maintained under controlled illumination (14 hr. light, 10 hr. darkness). Five rats of each genetic constitution were chosen; all had regular 5-day cycles. Blood samples were collected daily between 14.00 and 15.00 hr. from the occular venous plexus. The samples were used in the radioimmunoassays at the
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After the isolation from oestrogen-induced prolactin-producing tumours in mice of a protein (mouse pituitary tumour protein = MPTP), a radioimmunoassay has been developed based on 125I-labelled MPTP (labelled by the Chloramine-T method of Greenwood, Hunter & Glover, 1963) and a rabbit antiserum to MPTP suitable for the estimation of prolactin in pituitary tissue homogenates and mouse plasma (H. G. Kwa, F. Verhofstad & E. M. van der Bent, unpublished observations). Parallel inhibition curves obtained by (1) unlabelled MPTP, (2) mouse anterior lobe tissue and (3) plasma from pseudopregnant mice suggested that MPTP was prolactin. Results are now given for plasma levels of prolactin, expressed in mμg. of the MPTP standard, in female mice during the oestrous cycle and in μg. at various time intervals after implantation of isologous pituitary tissue under the renal capsule.
Blood samples were collected under light ether anaesthesia from the occular venous plexus, and the plasma
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SUMMARY
Many investigations of the regulation of prolactin synthesis and release are based on single plasma prolactin determinations. The purpose of the present experiment was to ascertain whether groups of rats (i.e. young or adult, male or female animals, being either intact, gonadectomized or gonadectomized and treated with oestrone), differing in age and/or endocrine status, will react to a single dose of perphenazine by an acute release of pituitary prolactin in proportion to their initial plasma prolactin levels.
No consistent relation existed between the classification of the twelve groups of rats into three categories of basal plasma prolactin levels (i.e. < 20, 25–50, > 125 ng/ml) and their response to perphenazine. Even though all groups showed a highly significant increase of plasma prolactin levels the magnitude of the maximum prolactin response at 30 min varied greatly within the groups of one category and thus was not related to the initial prolactin levels.
The effect of 14 days of oestrone treatment in increasing plasma prolactin levels in gonadectomized animals was greatest in young and adult male rats, less in young females and not significant in adult females. The results obtained after perphenazine treatment in the latter group made it clear that the effect of oestrogen treatment on prolactin release can be completely blocked by increasing synthesis and/or release of the prolactin-release inhibiting factor (PIF). Since perphenazine induces decrease of pituitary prolactin and a concomitant increase of plasma prolactin levels through lowered PIF-action, the positive effect of oestrogens on prolactin release (as observed in gonadectomized male and young female rats) apparently is caused by a different mode of action. The implications of these findings for the regulation of prolactin release, as affected by the endocrine status of the rat, is discussed.
Moreover, comparison of prolactin lost from the pituitary and gained in the circulation of the experimental animals, with amounts of prolactin that were observed to disappear from plasma during the experiment, provided suggestive evidence that the capacity to synthesize and/or eliminate prolactin, after a sudden provoked release of the hormone, differed among the groups.
The rates of synthesis by the pituitary, of release from the pituitary into the circulation as well as of elimination of the hormone from the circulation (equally involved in determining actual plasma levels) are thought, therefore, to be far more important for the elucidation of prolactin regulation than single plasma prolactin determinations.
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SUMMARY
l-DOPA, within 30 min after administration, induced a highly significant decrease of plasma prolactin levels (phase 1) in a number of groups of rats, differing in age and/or endocrine status, apparently by direct inhibition of prolactin release from the pituitary. Three hours after administration of l-DOPA these low plasma prolactin concentrations in treated animals had increased (phase 2) and did not differ significantly from levels in control animals, indicating that the effect of l-DOPA on plasma prolactin levels is only of short duration. During this process some interesting phenomena were observed, especially in the animals treated with oestrone. The elimination rate of prolactin from plasma was very high (t½ = 2·8 min), as indicated by decreasing concentrations of the hormone during phase 1. Pituitary prolactin content did not change during phase 1, suggesting that prolactin synthesis was also stopped. Notwithstanding the high elimination rate, plasma prolactin regained initial concentrations in phase 2, suggesting release of a substantial part of the pituitary prolactin content. The latter, however, remained constant during the whole experiment (i.e. before l-DOPA administration and during phase 1 as well as phase 2).
The results suggested another working mechanism of l-DOPA in decreasing plasma prolactin levels, namely by stimulating the uptake of this hormone in the periphery. After the effect of l-DOPA had ceased, most of the prolactin from the periphery returned into the bloodstream, causing a rapid restoration of plasma prolactin levels without substantial release from the pituitary. The nature of the processes responsible for the peripheral uptake of prolactin is discussed.
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In a study undertaken to try to correlate plasma levels, estimated by radioimmunoassay, and morphological signs of hyperactivity of the prolactin cells, three groups of spayed female rats bearing pituitary isografts were studied. These received (a) no oestrogen, (b) a low dose of 2 mg oestrone/l drinking water (0·5 mg/l being the dose that is sufficient to give vaginal cornification in spayed female rats), or (c) a high dose of 50 μg oestrone in oil/day (sufficient to induce pituitary tumours in rats). Varying degrees of prolactin secretory activity of both isografts and pituitary glands in situ were found, as shown by means of electron microscopy by the development of the endoplasmic reticulum, the Golgi region, and the number of secretory granules. The morphological signs of secretory activity correlated with the dosage of oestrogen, but plasma levels of prolactin in the rats without oestrogen treatment did not differ significantly from those
