Lydia A. Arbogast

Neuroendrocrine regulation of prolactin release

Article de synthese sur la regulation neuroendocrinienne de la liberation de la prolactine. Effet de la dopamine. Importance physiologique des facteurs de liberation de la prolactine

Progesterone reverses the estradiol-induced decrease in tyrosine hydroxylase mRNA levels in the arcuate nucleus

Estradiol (E2) and progesterone (P4) interact to influence tuberoinfundibular dopaminergic neuronal activity and contribute to the control of prolactin (PRL) release. This study examined tyrosine hydroxylase mRNA signal levels in the arcuate nucleus of the hypothalamus and tyrosine hydroxylase activity in the stalk-median eminence after 1 week of steroid treatment and related these to circulating PRL levels. Ovariectomized rats were untreated (control) or were implanted with E2, P4 or both E2 and P4 pellets and were sacrificed after 7 days at either 10:00 or 18:00 h. Some E2 + P4-treated rats were injected with either RU 486 or its vehicle at 12-hour intervals for the last 3 of the 7 days of steroid treatment. Tyrosine hydroxylase mRNA signal levels in the arcuate nucleus were decreased by 70% at both 10:00 and 18:00 h in the E2-treated rats compared to control rats. P4 alone had no effect on tyrosine hydroxylase mRNA levels, but reversed the E2-induced decrease so that mRNA levels in the E2 + P4-treated rats were similar to control levels. The progesterone antagonist RU 486 blocked this effect of P4, supporting the observation of decreased mRNA levels in E2-treated rats. Steroid treatment had no effect on tyrosine hydroxylase mRNA levels in the medial zona incerta. Tyrosine hydroxylase activity in the stalk-median eminence was similar at 10:00 and 18:00 h in control rats, and was decreased by 25 and 36% at 10:00 and 18:00 h, respectively, in E2-treated rats. P4 alone had no effect on tyrosine hydroxylase activity, but reversed the E2-induced decrease in enzyme activity to control levels at both 10:00 and 18:00 h. In contrast to the effect of RU 486 on tyrosine hydroxylase mRNA, tyrosine hydroxylase activity in E2 + P4-treated rats was not significantly altered by RU 486 at either 10:00 or 18:00 h. Circulating PRL levels were elevated in the E2-treated and E2 + P4-treated rats. A diurnal PRL rise was evident at 18:00 h in E2-treated rats, but was abolished by concomitant treatment with P4. The diurnal PRL surge was re-established in E2 + P4-treated rats after administration of RU 486, whereas basal circulating PRL levels were not altered by RU 486. These data indicate that P4 antagonizes the profound inhibitory effect or E2 on tyrosine hydroxylase mRNA content in the tuberoinfundibular dopaminergic neurons.(ABSTRACT TRUNCATED AT 400 WORDS)

Ontogeny of tyrosine hydroxylase mRNA signal levels in central dopaminergic neurons: development of a gender difference in the arcuate nuclei

Using in situ hybridization and immunocytochemistry, this study examined the tyrosine hydroxylase (TH) mRNA signal levels and immunostaining in the arcuate nuclei of the hypothalamus, zona incerta and substantia nigra of male and female rats during neonatal, peripubertal and adult life. The catalytic activity of TH in the stalk-median eminence was also investigated using the in vitro accumulation of 3,4 dihydroxyphenylalanine (DOPA) after inhibiting aromatic amino acid decarboxylase activity. In the arcuate nuclei, TH mRNA levels increased 3.5-fold between 5 and 15 days of age and remained at a steady level between 15 and 35 days of age in both male and female rats. Similar to TH mRNA levels in the arcuate nuclei, TH activity in the stalk-median eminence increased 2-3 fold between days 10 and 15 of age and remained at a steady level between 15 and 35 days of age in both sexes. A later increase in TH mRNA levels in the arcuate nuclei and catalytic activity in the stalk-median eminence was observed between 35 and 40 days in females, but not males. During adulthood, TH mRNA levels and enzyme activity were 2.7-fold higher in the arcuate nuclei and stalk-median eminence, respectively, of diestrous females vs males. These data suggest that the changes in TH mRNA in the arcuate nuclei may contribute to the developmental alterations, as well as the adult gender differences, in enzyme activity in the stalk-median eminence. Circulating progesterone levels were low (1-10 ng/ml) between days 5 and 25 of age and increased 6-fold between 25 and 35 days of age in both males and females. Progesterone levels increased 2-fold in females, but not males, between days 35 and 40 and were 4-fold higher in diestrous females as compared to adult males. Circulating prolactin levels were low (2-3 ng/ml) between days 5 and 15, increased 15-fold between days 15 and 25 and increased an additional 2- to 3-fold between days 35 and 70 in both males and females. TH mRNA signal levels increased between days 5 and 15 of age in dopaminergic perikarya in the zona incerta and the substantia nigra of both female and male rats. The TH mRNA levels remained constant between days 15 and 70 in the zona incerta, whereas TH mRNA levels declined with age in the substantia nigra.(ABSTRACT TRUNCATED AT 400 WORDS)

Chapter 12 Regulation of prolactin secretion during pregnancy and lactation

Abstract Prolactin plays major roles in maintaining the corpora lutea of pregnancy and in the synthesis of milk during lactation. The hypothalamic mechanisms involved in these functions have been investigated. Mating leads to a surge of prolactin and programs daily surges during early pregnancy. The expression of Fos-immunoreactivity shows that mating activates several hypothalamic nuclei, particularly the arcuate nucleus and medial preoptic area. In the arcuate nucleus, mating is associated with Fos expression in β-endorphin neurons, and infusion of naloxone blocks both mating-induced and diurnal prolactin surges. Tyrosine hydroxylase-immunoreactive dopamine neurons appear not to participate in surge generation. However, after day 10 of gestation the secretion of placental lactogens suppresses prolactin secretion via activation of dopamine neurons without involvement of β-endorphin neurons. Intracerebroventricular implantation of placental lactogen-secreting cells will block pregnancy prolactin surges, increase Fos expression in dopamine neurons, and increase tyrosine hydroxylase activity. During lactation the mechanisms regulating dopamine and β-endorphin neurons are further modified. In early lactation a prolactin-induced increase in tyrosine hydroxylase activity leads to negative feedback, but this effect is lost by mid-lactation. Overriding this negative feedback is the inhibitory effect that suckling has on dopaminergic activity. This may involve β-endorphin-mediated inhibition of dopamine neurons, as naloxone causes a marked increase in tyrosine hydroxylase activity and suppression of circulating prolactin. However, removal of tonic dopamine inhibition is not sufficient to account for the high levels of prolactin attained during lactation, and additional releasing factors are probably involved. In situ hybrization histochemistry for the most recent candidate, prolactin-releasing peptide, suggests that this may involve brain stem neurons that co-localize noradrenaline. Thus, prolactin secretion during pregnancy and lactation involve complex interactions of regulatory factors and plasticity of neuronal responsiveness.

Estradiol Attenuates the Forskolin-Induced Increase in HypothalamicTyrosine Hydroxylase Activity

The purpose of this study was to evaluate interactions between estradiol and the 3′,5′ cyclic adenosine monophosphate (cAMP) signaling pathway to regulate tyrosine hydroxylase (TH) activity in hypothalamic dopaminergic neurons. The first experiment examined the ability of forskolin to activate TH in the tuberoinfundibular dopaminergic neurons of adult ovariectomized rats with or without estradiol treatment. Estradiol treatment reduced both basal and forskolin-stimulated TH activity in the median eminence. The second group of experiments examined the effect of estradiol on the forskolin-induced activation of TH in fetal hypothalamic cells cultures. Estradiol decreased basal TH activity in the hypothalamic cell cultures to 80% of control levels. Forskolin treatment for 1 h increased TH activity in a concentration-dependent manner in control and estradiol-treated cells, but estradiol attenuated the stimulatory response to 0.01–10 µM forskolin. The suppressive effect of estradiol on cAMP-dependent activation of TH was evident with 1–12 h of forskolin treatment. The responses to other activators of the cAMP- protein kinase A pathway, including dibutyryl cAMP and 8-bromo-cAMP, and to a depolarizing stimulus were blunted in estradiol-treated cultures. Forskolin treatment for 1 h increased radiolabeled phosphate incorporation into TH protein in control but not estradiol-treated cells, suggesting that estradiol interferes with the ability of the cAMP pathway to phosphorylate TH. Forskolin caused a time-dependent increase in TH mRNA signal levels in control cultures. The magnitude of the forskolin-induced increase in TH mRNA levels was less in the estradiol-treated cells after 6 h of forskolin treatment, indicating that estradiol hinders cAMP-regulated TH gene expression. These data indicate that estradiol attenuates the ability of hypothalamic dopaminergic neurons to respond to cAMP-dependent stimulation by interfering with phosphorylation mechanisms in the short term and control of TH mRNA levels in the long term.

Tyrosine hydroxylase messenger RNA in the hypothalamus, substantia nigra and adrenal medulla of old female rats

The effects of aging in the female rat were analyzed in terms of tyrosine hydroxylase (TH) gene expression and serum prolactin levels. The number of tuberoinfundibular dopaminergic (TIDA) neurons and the concentration of TH mRNA per cell was greater in 16- to 18-month-old rats than in 25-month-old rats. The amount of TH immunostaining was more intense in the median eminence of the 18-month-old rats compared to either younger or older rats. Plasma prolactin levels were moderately elevated in 18-month-old rats compared to 4-month-old rats, and extremely elevated in 25-month-old rats due to the occurrence of pituitary prolactinomas. There were no detectable changes in TH mRNA levels in the substantia nigra with age, whereas adrenal TH mRNA increased with age. We propose that prolactin initially exerts a stimulatory effect on the TIDA neurons as the rat ages, but eventually causes a loss in neuronal number and neuronal function as the pituitary prolactinoma secretes increased amounts of prolactin.

Gene Expression Profiles of Intracellular and Membrane Progesterone Receptor Isoforms in the Mediobasal Hypothalamus During Pro-Oestrus

Progesterone action is mediated by its binding to specific receptors. Two progesterone receptor (PR) isoforms (PRA and PRB), three membrane progesterone receptor (mPR) subtypes (mPRα, mPRβ and mPRγ) and at least one progesterone membrane‐binding protein [PR membrane component 1 (PRmc1)] have been identified in reproductive tissues and brain of various species. In the present study, we examined gene expression patterns for PR isoforms, mPR subtypes and PRmc1 in the rat mediobasal hypothalamus (MBH) during pro‐oestrus. The mRNA level for each receptor subtype was quantified by a real‐time reverse transcriptase‐polymerase chain reaction (RT‐PCR) at the time points: 13.00 h on dioestrous day 2; 09.00, 13.00, 17.00 and 22.00 h on pro‐oestrus; and 13.00 h on oestrus. For PR, one primer set amplified PRA+PRB, whereas a second primer set amplified PRB. As expected, PRA+PRB mRNA expression was greater than PRB in MBH tissue. PRB mRNA levels increased throughout the day on pro‐oestrus, with the highest levels being observed at 17.00 h. PRB mRNA levels in the MBH were increased by 2.4‐ and 3.0‐fold at 13.00 and 17.00 h, respectively, on pro‐oestrus compared to 13.00 h on dioestrous day 2. There were differential mRNA expression levels for mPRs and PRmc1 in the MBH, with the highest expression for PRmc1 and the lowest for mPRγ. The mPRα mRNA contents at 13.00 and 17.00 h on pro‐oestrus were increased by 1.5‐fold compared to that at 13.00 h on dioestrous day 2. The mPRβ mRNA levels at 13.00 and 17.00 h on pro‐oestrus were 2.5‐ and 2.4‐fold higher compared to that at 13.00 h on dioestrous day 2, respectively. PRA+PRB, mPRγ and PRmc1 mRNA levels did not vary on pro‐oestrus. These findings suggest that the higher expression of PRB, mPRα and mPRβ in the MBH on pro‐oestrous afternoon may influence both genomic and nongenomic mechanisms of progesterone action during the critical pre‐ovulatory period.

Differential Alterations in Dopamine Turnover Rates in the Stalk-Median Eminence and Posterior Pituitary during the Preovulatory Prolactin Surge

The relative contributions of dopamine (DA) and prolactin-releasing factor (PRF) in generating the preovulatory prolactin (PRL) surge were investigated. Immature female rats were injected with pregnant mares serum gonadotropin (PMSG) on day 28. Jugular blood was collected hourly on days 30 and 31. PRL levels were low in the morning of day 30, rose 10-12 times to peak levels from 14.00 to 16.00 h, reached a prolonged plateau from 18.00 to 24.00 h, and reduced to basal levels in the morning of day 31. All PMSG-treated rats ovulated an average of 13-14 ova. PRL levels in age-matched control rats were low throughout this time, and no oviductal ova were present. DA turnover rates in the stalk-median eminence (SME) and posterior pituitary (PP) were determined from the decline in tissue DA after injecting alpha-methyl-p-tyrosine (alpha-MPT), a competitive inhibitor of tyrosine hydroxylase. DA turnover rates increased or were unaltered in the SME and PP, respectively, during the peak PRL phase as compared to presurge rates. In contrast, DA turnover rates were significantly reduced in both tissues during the plateau phase. The turnover rate in the SME, but not the PP, was increased in the morning of day 31. DA turnover rates in control rats never changed. Injection of alpha-MPT to PMSG-treated rats increased PRL levels at all times examined except during the plateau phase. Blood PRL levels were also determined in PMSG-treated rats following posterior pituitary lobectomy or sham lobectomy. The PRL surge was similar in both groups and all rats ovulated.(ABSTRACT TRUNCATED AT 250 WORDS)

Progesterone Induces Dephosphorylation and Inactivation of Tyrosine Hydroxylase in Rat Hypothalamic Dopaminergic Neurons

After in vivo treatment, progesterone initially decreases tyrosine hydroxylase (TH) activity in the TIDA neurons, but subsequently increases TH activity with prolonged treatment. In order to explore the cellular mechanism for progesterone’s effect, this study examined the acute inhibitory action of progesterone on TH activity in rat fetal hypothalamic dopaminergic neurons in vitro. Progesterone caused a rapid decrease in TH activity within 1 h, which was sustained for at least 6 h. However, the dopaminergic cells became refractory to progesterone with continuous treatment for 12 h to 10 days. Progesterone (10–100 nM) treatment suppressed TH activity in a concentration-dependent manner. The inhibitory effect of progesterone was dependent on prior exposure to estradiol. Whereas progesterone decreased TH activity, A ring-reduced metabolites of progesterone did not alter TH activity, suggesting that the response was specific to progesterone. Progesterone decreased radiolabeled phosphate incorporation into TH protein. Okadaic acid, a phosphoprotein phosphatase inhibitor, prevented the progesterone-induced suppression of TH activity and phosphate incorporation into TH, implicating dephosphorylation of TH as the cellular mechanism. In contrast, neither TH mRNA levels nor TH protein content was altered after 1 or 12 h of progesterone treatment. Progesterone decreased TH activity after pretreatment of the hypothalamic cells for 2 or 24 h with actinomycin D, an RNA synthesis inhibitor, suggesting that increased transcription does not mediate the effect. These data indicate that the acute progesterone-induced decline in TH activity is caused by dephosphorylation of TH.

Semicircadian rhythms of c-Fos expression in several hypothalamic areas during pregnancy in the rat: relationship to prolactin secretion.

Prolactin (PRL) serves an important luteotrophic function in the rat during early pregnancy, expressed as a nocturnal surge in the early morning and a diurnal surge in the late afternoon. Several areas of the hypothalamus, including the preoptic area (POA), the suprachiasmatic nucleus (SCN) and the ventromedial and dorsomedial nuclei (VM-DM) have been implicated in PRL surges. We investigated the temporal relationship between neuronal activity as measured by c-Fos immunocytochemistry in these areas and PRL secretion during early and late pregnancy. Brains were collected at nine time points (24:00, 02:00, 04:00, 06:00, 10:00, 14:00, 16:00, 18:00 and 20:00 h) on days 6–7 and three time points (02:00, 14:00 and 18:00 h) on days 14–15 of pregnancy. Plasma PRL levels determined by radioimmunoassay revealed two surges with peaks at 02:00 and 18:00 h and a trough at 14:00 h on days 6–7, which were absent on days 14–15 of pregnancy. The number of neurons expressing c-Fos in the anterior medial preoptic nucleus, the medial preoptic area and the medial preoptic nucleus, but not the anteroventral preoptic nucleus of the POA, and the VM-DM, showed a semicircadian rhythm which was maximal at 02:00 h or/and 04:00 and 18:00 h and reached the lowest value at 14:00 h, in parallel with the PRL surges in early pregnancy. However, the temporal pattern of c-Fos in these areas was reversed during late pregnancy, with a peak at 14:00 h and low levels at 02:00 and 18:00 h. PRL surges were absent and levels were uniformly low during these times. Neuronal activity in the SCN did not show any correlation with PRL surges. The dorsomedial subdivision of the SCN showed high neuronal activity during the daytime in both stages of pregnancy. Neuronal activity in the ventrolateral subdivision of the SCN was high during the nighttime in early pregnancy, however it exhibited high levels during the daytime in late pregnancy. These results suggest that the two daily surges of PRL secretion during the first half of pregnancy might be related to the temporal rhythm of neuronal activity in the POA and the VM-DM, and a major change in the pattern of neuronal activity in these hypothalamic areas might result in termination of the PRL surges at midpregnancy.