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ABSTRACT
The autoregulation of rat prolactin secretion at the level of the pituitary gland was investigated, using a static incubation system. The rate of prolactin secretion from the female anterior pituitary gland in vitro was found to be constant when the medium was changed at 20-min intervals. However, when the medium was left unchanged and secretory products were allowed to accumulate, prolactin secretion began to decline within 60 min. This effect was not observed with the male tissue, where the level of accumulated prolactin did not reach that at which the inhibition occurred using female tissue.
The nature of the putative secretory product causing the inhibition of prolactin secretion was investigated. Exogenous bovine prolactin (1–4 mg/l) caused an inhibition of endogenous rat prolactin secretion. Inclusion of monoamine oxidase in unchanged medium, to prevent dopamine accumulation in the medium (a possible consequence of co-storage and cosecretion with prolactin granules), did not prevent the inhibition observed in the control incubation. We therefore conclude that in-vitro autoregulation of prolactin secretion can occur at the level of the pituitary gland, probably due to the accumulated prolactin having a feedback action on the lactotroph. This might be of physiological significance if localized concentrations of the hormone within the gland are high.
J. Endocr. (1987) 115, 13–18
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ABSTRACT
Experiments were carried out on the antagonistic effects of opiates on the inhibition by dopamine of prolactin secretion from rat anterior pituitary glands. Dose–response and time-course experiments were carried out using both static incubation of paired hemipituitary glands and perifusion of whole glands. Dopamine (10–1000 nmol/l) was found to have an inhibitory effect on prolactin secretion, but at a lower concentration (0·1 nmol/l) a small stimulation was observed. Against an inhibition established with 100 nmol dopamine/l in static incubation, the three opiates under study, morphine sulphate, Leu5enkephalin and d-Ala2,Met5-enkephalin (DAME), had a maximum antagonistic effect at 50–1000 nmol/l in a 90-min incubation. Morphine and DAME were rather more effective than Leu5-enkephalin, possibly because of degradation of the latter. Naloxone reversed the effect of morphine. All three opiates showed little effect on dopamine-inhibited prolactin secretion in a perifusion system. The data accord with previous suggestions that prolactin secretion may be stimulated both by very low concentrations of dopamine and by opiates acting to reverse the inhibition exerted by higher dopamine concentrations. It should be noted that both morphine and the enkephalins have similar effects on prolactin secretion, despite their normal specificity for different opiate receptors; their actions on the pituitary may thus be rather non-specific.
J. Endocr. (1986) 109, 313–320
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The effects of domperidone on prolactin secretion from the anterior pituitary glands of female rats were studied. A perifusion system and radioimmunoassay were used to study secretion; static incubations and incorporation of [3H]leucine were used to measure biosynthesis. During perifusion, intact anterior lobes showed a constant rate of prolactin secretion for up to 5 h (after a 90-min preincubation to stabilize the tissue). Incorporation studies revealed an increase in protein synthesis in perifused hemipituitary glands. When glands were treated with 10 nm-domperidone, prolactin secretion began to decline after the first hour, reaching a maximum of 40–50% inhibition after a further 90 min. Growth hormone secretion showed no such decline. Inhibition of prolactin secretion continued for up to 2 h after withdrawal of the drug. Treatment with 100 nm-dopamine resulted in a more rapid inhibition of secretion, but the effect was reversed on withdrawal of the catecholamine. The prolactin content of perifused hemipituitary glands was measured after treatment with domperidone; the contents of control and treated glands did not differ, but were depleted compared with hemipituitary glands which had not been perifused.
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Anterior pituitary glands from female rats were dispersed enzymically in the absence of dopamine. Dispersed cells (106–107) were layered onto columns containing Bio-Gel P-2 and were then perifused for 3 h with Dulbecco's Modified Eagle's Medium. The prolactin content of the perifusate and cell homogenates was determined by radioimmunoassay. Prolactin secretion during the third hour of perifusion increased as the loading of cells increased. However, the increase was not linear, and when secretion rate per 106 cells was calculated it was found that increased loading decreased the rate, which fell to a plateau of 1·3 ± 0·1 (s.e.m.) ng/min per 106 cells at a loading of about 8 × 106 cells from 3·8 ± 0·1 ng/min per 106 cells for a loading of 106 cells. This cell-density dependence of the rate of prolactin secretion in the perifusion system may be due to intercellular contact since the isolation of the tissue removes the influence of hypothalamic factors, while localized build up of prolactin (potentially causing direct autoregulation on the lactotroph) seems unlikely because of the continuous flow of medium.
