Leu-enkephalin-stimulated growth hormone and prolactin release in the rat: comparison with the effect of morphine

The effect of Leu⁵-enkephalin on growth hormone (GH) and prolactin (PRL) release was studied $\text{in vivo}$ in the infant rat and compared to that of morphine. In 10 day-old pups, intracerebroventricular injection of Leu⁵-enkephalin (50, 75 and 100 μg) resulted in a dose-related increase in plasma GH; morphine was active as GH releaser at the dose of 5 and 10 μg, but not at 2.5 μg. Pretreatment with naloxone (2 mg/kg ip) suppressed the GH-releasing effect of either Leu⁵-enkephalin (100 μg) or morphine (10 μg). Leu⁵-enkephalin (75 and 100 μg) induced a rise in plasma PRL which was neither dose-related nor antagonized by naloxone; morphine (5 and 10 μg) was active as PRL releaser and its effect was antagonized by naloxone. These results indicate that: 1) Leu⁵-enkephalin stimulates both GH and PRL release; 2) the release of GH by Leu⁵-enkephalin but likely not that of PRL involves specific opiate receptors; 3) morphine releases GH and PRL through specific opiate receptors.     

Ontogeny of the opioidergic regulation of LH and prolactin secretion in lactating sows. II: Interaction between suckling and morphine administration

Administration of morphine to ten suckled and nine zero-weaned (piglets removed immediately after farrowing) sows was used to investigate the apparent absence of opioid regulation of LH and prolactin secretion in early lactation. Blood samples were collected at 10 min intervals at 24-30, 48-54, 72-78 h post partum, and for a 12 h period from 08:00 to 20:00 on day 10 after farrowing. Morphine (0.1 mg kg-1) was administered as three i.v. bolus injections at intervals of 1 h during the last 3 h of each of the 6 h sampling periods, and at 6, 7 and 8 h after the beginning of sampling on day 10. There were significant (P < 0.001) group (zero-weaned versus suckled), time and morphine effects on LH secretion. Plasma LH concentrations increased (P < 0.001) within 48 h of farrowing in zero-weaned sows. Long-term trends of an increase in mean plasma LH in the sampling periods before treatment were attenuated in both groups by morphine treatment. Morphine also significantly inhibited (P < 0.05) prolactin secretion in suckled sows. In zero-weaned sows, plasma prolactin was already low at the start of sampling and did not change with time or in response to morphine treatment. Therefore, the inability to demonstrate an opioidergic involvement in the suckling-induced inhibition of LH secretion during the early post-partum period in sows is not due to a lack of opioid receptors. Furthermore, in suckled sows, morphine is stimulatory to systems that have an inhibitory effect on prolactin secretion.     

Ultrasonic cries from infant rats stimulate prolactin release in lactating mothers

Lactating female rats, separated from their litters for 6 hr, showed a dramatic elevation in plasma prolactin following a 15-min playback of ultrasonic vocalizations recorded from 7-day-old pups. Virgin females exhibited a smaller but still significant prolactin response to recorded pup calls. Control recordings of either background noise or adult ultrasonic vocalizations (22–26 kHz) had no effect on prolactin secretion even in lactating females. These results demonstrate that infant vocalizations can act as a stimulus for prolactin release and suggest that such communications may play a role in the maintenance of normal lactation.     

Gonadotropin and prolactin secretion following intraventricular administration of morphine in gilts

The effects of central nervous system administration of morphine on secretion of luteinizing hormone (LH), follicle-stimulating hormone, and prolactin were investigated in ovariectomized gilts stereotaxically implanted with lateral ventricular cannulas. In Experiment 1, mean serum LH and follicle-stimulating hormone concentrations and serum LH pulse frequency were unaffected by artificial cerebrospinal fluid administration (P greater than 0.1), but decreased (P less than 0.01) in 8 of 11 gilts when 500 micrograms of morphine were given 3 hr later. Serum LH pulse amplitude was unaffected (P greater than 0.1) by cerebrospinal fluid or morphine injection. In Experiment 2, luteinizing hormone concentrations decreased (P less than 0.0001) and prolactin concentrations increased (P less than 0.0001), but follicle-stimulating hormone concentrations did not change (P greater than 0.1) after 500 micrograms of morphine. Gonadotropin responses to 10 micrograms of gonadotropin-releasing hormone, given 2 hr after intraventricular injection, were similar (P greater than 0.1) for morphine- and cerebrospinal fluid-treated gilts. These results indicate that morphine inhibits LH secretion at the level of the central nervous system, and are consistent with the concept that endogenous opioid peptides participate in the regulation of gonadotropin and prolactin release in pigs.     

Pulsatile secretion of prolactin and oxytocin during nursing in the lactating rat

The secretory profile of prolactin and oxytocin in response to suckling stimuli by litters was studied in unanesthetized and urethane-anesthetized lactating rats. Serum prolactin levels were determined by radioimmunoassay. Oxytocin released at milk-ejection reflex was monitored by the changes in the intramammary pressure and/or the characteristic pup's reaction associated with the milk ejection. Serum prolactin concentrations began to rise earlier than the first milk ejection in unanesthetized rats, but they were never elevated without the appearance of milk ejections in urethane-anesthetized rats. Pulsatile fluctuation in serum prolactin levels at 6-15 min intervals was observed in the nursing period when 10 pups were suckling continually. The intermittent milk-ejection reflex occurred not always but preponderantly (64-91%) when the serum prolactin levels were at the nadir of the fluctuation. Injection of an estimated dose of oxytocin released at each milk ejection (1 mU) did not change the serum prolactin levels. These results indicate that the mechanism for prolactin release may be more susceptible to the effects of anesthesia than that for oxytocin release in response to the suckling stimuli and that the release of both the hormones is pulsatile in nature and be influenced by a common biological clock during the nursing period.     

Extensive ultrastructural changes in rat mammotrophs following administration of the dopamine agonist ergocristine-reflecting inhibition of prolactin release

Summary We have demonstrated an extensive reorganization of organ elles in mammotrophs immediately following administration of ergocristine (a dopamine agonist) to estradiol-primed male rats. Our ultrastructural findings are consistent with our previous results that ergocristine can block prolactin release without any noticeable latent period. Following three-week priming of male rats with estradiol implants, ergocristine was administered by a bolus injection through an indwelling cannula. Within two min of its administration, ergocristine induced dramatic changes in the ultrastructure of mammotrophs, i.e., (1) increased numbers of secretory granules, (2) peripheral relocation of rough endoplasmic reticulum which tends to sequester secretory granules, (3) change in location of nucleus and (4) increased numbers of intracellular bodies associated with secretory granules. We suggest that the extensive ultrastructural changes that occurred in such a short period following ergocristine administration may be indications of specific factors associated with blockage of hormone release.     

Central adrenoceptors and basal prolactin release in the rat

The influence of the central adrenergic system on basal prolactin secretion was investigated in the rat. Several selective adrenoceptor blockers were centrally administered and their effects on prolactin secretion were observed. Blockade of beta-1 receptors by practolol, beta-2 receptors by IPS 339 and alpha-2 receptors by DG-5128 did not modify basal prolactin secretion, but alpha-1 adrenoceptor blockade by prazosin strongly enhanced prolactin plasma levels. These findings suggest that noradrenergic pathways in the central nervous system exert inhibitory tone on basal prolactin secretion, and that this effect seems to be mediated by alpha-1 adrenoceptors.     

Proteolytic modification of rat prolactin by subcellular fractions of the lactating rat mammary gland

The current study explored prolactin proteolysis by rat lactating mammary gland. 125I-labelled rat prolactin was incubated with tissue fractions of lactating mammary gland and the extent of prolactin degradation and fragment formation was visualized and densitometrically quantitated from autoradiographs derived from SDS-polyacrylamide gel electrophoresis under reducing conditions. At pH 4.5, the 25 000 X g pellet of mammary gland converted intact prolactin (23 kDa band) to proteolytic fragments (8-16 kDa bands) in a time- and tissue concentration-dependent fashion similar to that reported previously for rat ventral prostate. The prolactin-degrading and -fragmenting activity in lactating mammary gland was 5-10-times that observed for ventral prostate, the most active male tissue. This activity at acid pH was also demonstrable in other fractions of mammary gland but appeared to predominate in the cytosol. The above activities in mammary gland virtually disappeared at pH 7.4, appeared sensitive to aspartate and sulfhydryl proteinase inhibitors, and insensitive to serine and metalloenzyme proteinase inhibitors. The distribution of this activity could not be correlated with a particular enzyme marker. These characteristics of mammary gland activity differed significantly from those reported previously for prostate. When electrophoresis was conducted under non-reducing conditions, prolactin proteolysis in prostate and mammary gland was primarily associated with the formation of a more slowly migrating product (24 kDa band) with little spontaneous 8-16 kDa fragment formation. Re-electrophoresis of the 24 kDa band under reducing conditions resulted in the appearance of the 8 and 16 kDa fragments. In conclusion, prolactin is proteolytically modified by prostate and lactating mammary gland to a variant of intact hormone (24 kDa band) with a cleavage site in its large loop, by two or more widely distributed, acid-dependent proteinases. Lactating mammary gland, the principal target for prolactin, has the capacity to cleave the hormone in its loop at rates higher than any other tissue examined to date.     

The influence of testosterone administration on plasma prolactin levels in the neonatally androgenized (NA) female rat

Neonatally androgenized (NA) female rats were ovariectomized (OVX) as adults and given 1 mg of testosterone propionate/day for 7 days and the plasma prolactin (PRL) pattern compared with NA intact animals and normal OVX animals given estrogen or TP. NA intact animals had elevated basal (morning values) and an attenuated afternoon surge when compared to normal estrogen-treated animals. Testosterone administration to normal animals induced an afternoon surge similar to that of normal estrogen-treated animals but the magnitude of the surge was less. Testosterone given to NA-OVX animals had little effect on either morning or afternoon PRL levels. The results suggest that in the NA rat the brain region involved in the conversion of testosterone to estrogen may be altered by neonatal androgen exposure.     

Interference with prolactin release and the maternal behavior of female rats

The role of prolactin in maintaining the maternal behavior of the rat was investigated. Ergocornine, a drug that interferes with the release of prolactin from the anterior pituitary, was injected daily into postpartum females for a period of 21 days. Those receiving ergocornine did not differ significantly from control females on any measure of maternal behavior, the control females having received either ergocornine plus prolactin or simply the vehicle. Since both lactation and the decidual cell response to uterine traumatization were inhibited after ergocornine, and since each were at least partially reversed by exogenous prolactin, it was evident that ergocornine interfered successfully with the postpartum release of prolactin. That at the same time maternal behavior was unaffected, supports the conclusion that prolactin is not essential in maintaining the maternal responsivity of the postpartum female rat.     

Time course of the ventilatory response to adjuvant arthritis in the rat

The present study determined the time course of the ventilatory response to adjuvant arthritis in the rat. It was found that the minute ventilation of arthritic rats reliably exceeded that of control animals during a period of about 4 weeks. The ventilatory response had its onset during the 2nd week after inoculation of $\text{Mycobacterium butyricum}$ in the tail base, peaked on day 21, and was statistically reliable up to the 35th day. A smaller difference persisted throughout the further six weeks of the study, which covered 77 days in all. The changes in minute ventilation appeared to be closely time-locked with the changes in body weight and paw diameter which are characteristic of rats with adjuvant arthritis. The peak minute ventilation of individual animals also correlated in a significant manner with changes in weight and in paw diameter. The data are consistent with an earlier attempt to characterize the time course of the chronic pain which is presumably associated with adjuvant arthritis in the rat; minute ventilation is discussed as a possible measure of this putative pain.     

Possible role of vasoactive intestinal polypeptide on prolactin release during suckling in lactating women

The effect of suckling on plasma vasoactive intestinal polypeptide (VIP) values was evaluated in 6 nursing women on the 3rd to 4th day postpartum. Plasma prolactin (PRL) concentrations were also measured. Plasma VIP values (20.6 +/- 3.2 pg/ml in baseline) significantly (p less than 0.05) increased, reaching a maximum (53.5 +/- 10.9 pg/ml) 20 min after the starting of suckling. A possible role of VIP in the suckling-induced PRL release in humans cannot be excluded.     

Prolonged morphine administration alters protein expression in the rat myocardium

Background  

Morphine is used in clinical practice as a highly effective painkiller as well as the drug of choice for treatment of certain heart diseases. However, there is lack of information about its effect on protein expression in the heart. Therefore, here we aimed to identify the presumed alterations in rat myocardial protein levels after prolonged morphine treatment.     

Effects of enkephalin in analogues on prolactin release in the rat.

The effects of nineteen enkephalin analogues on the circulating levels of prolactin in the male rat following intraventricular injection of the peptides were determined and compared with that of Met- and Leu-enkephalin. Eleven of the 19 analogues stimulated prolactin secretion. It was found, in general, that the structure activity relationship for enkephalin stimulation of prolactin secretion was similar to that for opiate receptor activity. Analogues which contained a [DAla²] substitution were generally effective in stimulating prolonged prolactin release. Some, but not all analogues containing [DTrp²] or [DLeu⁵] were active. Analogues containing the [DTrp¹], [DPhe⁴] or [DMet⁵] substitutions were ineffective. The prolactin releasing effect of intravenous Tyr-DAla-Gly-Phe-DLeu was reversed by naloxone. Naloxone had no effect on the haloperidol- and alpha-methylparatyrosine induced increases in plasma prolactin levels. The results of these studies are discussed in the light of the suggestion that the enkephalins may function as neuroendocrine modulators.     

PMA-sensitive protein kinase C is not necessary in TRH-stimulated prolactin release from female rat primary pituitary cells.

In GH3 cells and other clonal rat pituitary tumor cells, TRH has been shown to mediate its effects on prolactin release via a rise of cytosolic Ca2+ and activation of protein kinase C. In this study, we examined the role of protein kinase C in TRH-stimulated prolactin release from female rat primary pituitary cell culture. Both TRH and PMA stimulated prolactin release in a dose-dependent manner. When present together at maximal concentrations, TRH and PMA produced an effect which was slightly less than additive. Pretreatment of rat pituitary cells with 10(-6) M PMA for 24 hrs completely down-regulated protein kinase C, since such PMA-pretreated cells did not release prolactin in response to a second dose of PMA. Interestingly, protein kinase C down-regulation had no effect on TRH-induced prolactin release from rat pituitary cells. In contrast, PMA-pretreated GH3 cells did not respond to a subsequent stimulation by either PMA or TRH. Pretreatment of rat pituitary cells with TRH (10(-7) M, 24 hrs) inhibited the subsequent response to TRH, but not PMA. Forskolin, an adenylate cyclase activator, stimulated prolactin release by itself and in a synergistic manner when incubated together with TRH or PMA. The synergistic effects of forskolin on prolactin release was greater in the presence of PMA than TRH. Down-regulation of protein kinase C by PMA pretreatment abolished the synergistic effect produced by PMA and forskolin but had no effect on those generated by TRH and forskolin. sn-1,2-Dioctanylglycerol (DOG) pretreatment attenuated the subsequent response to DOG and PMA but not TRH. The effect of TRH, but not PMA, on prolactin release required the presence of extracellular Ca2+. In conclusion, the mechanism by which TRH causes prolactin release from rat primary pituitary cells is different from that of GH3 cells; the former is a protein kinase C-independent process whereas the latter is at least partially dependent upon the activation of protein kinase C.