Prolactin induces a hyperpolarising current in rat paraventricular oxytocinergic neurones.

Prolactin and oxytocin are important reproductive hormones implicated in several common adaptive functions during pregnancy, pseudopregnancy and lactation. Recently, extracellular recordings of supraoptic neurones have shown that prolactin may modulate the electrical activity of oxytocinergic neurones. However, no study has been conducted aiming to establish whether prolactin directly influences this activity in oxytocinergic paraventricular neurones. In the present study, we addressed this question by studying the effects of prolactin on the electrical activity and voltage-current relationship of identified paraventricular neurones in rat brain slices. Whole-cell recordings were obtained and neurones were classified on the basis of their morphological and electrophysiological fingerprint (i.e. magnocellular or parvicellular) and neuropeptide phenotype (i.e. oxytocinergic or non-oxytocinergic). We report that prolactin elicited a hyperpolarising current in 37% of the neurones in this nucleus, of which the majority (67%) were identified as putative magnocellular oxytocin neurones and the reminder (33%) were regarded as oxytocin-negative, parvicellular neuroendocrine neurones. Our results suggest that, in addition to the well-established negative feedback loop between prolactin-secreting lactotrophs and dopaminergic neurones in the arcuate nucleus, an inhibitory feedback loop also exists between lactotrophs and oxytocinergic paraventricular neurones.

Investigating heterogeneity of intracellular calcium dynamics in anterior pituitary lactotrophs using a combined modelling/experimental approach.

Cell responses are commonly heterogeneous, even within a subpopulation. In the present study, we investigate the source of heterogeneity in the Ca(2+) response of anterior pituitary lactotrophs to a Ca(2+) mobilisation agonist, thyrotrophin-releasing hormone. This response is characterised by a sharp increase of cytosolic Ca(2+) concentration as a result of mobilisation of Ca(2+) from intracellular stores, followed by a decrease to an elevated plateau level that results from Ca(2+) influx. We focus on heterogeneity of the evoked Ca(2+) spike under extracellular Ca(2+) free conditions. We introduce a method that uses the information provided by a mathematical model to characterise the source of heterogeneity. This method compares scatter plots of features of the Ca(2+) response obtained experimentally with those made from the mathematical model. The model scatter plots reflect random variation of parameters over different ranges, and matching the experimental and model scatter plots allows us to predict which parameters are most variable. We find that a large degree of variation in Ca(2+) efflux is a likely key contributor to the heterogeneity of Ca(2+) responses to thyrotrophin-releasing hormone in lactotrophs. This technique is applicable to any situation in which the heterogeneous biological response is described by a mathematical model.

A tale of two rhythms: the emerging roles of oxytocin in rhythmic prolactin release.

Hormone secretion often occurs in a pulsatile manner. In this review, we discuss two rhythms of in vivo prolactin release in female rats and the ongoing research that we and others have performed aiming to understand the mechanisms underlying them. The peptide hormone oxytocin appears to play an important role in both rhythms. One rhythm occurs during the first half of pregnancy, but can also be induced in ovariectomised rats. This is characterised by a circadian pattern with two prolactin surges per day. Two methods for triggering this rhythm are discussed, each utilising a unique physiological pathway that includes oxytocin action, presumably on pituitary lactotrophs. The second rhythm occurs during the oestrous cycle and is characterised by a surge of prolactin on the afternoon of pro-oestrus. We discuss recent findings that oxytocin is more effective at stimulating prolactin release from lactotrophs taken from animals on the afternoon of pro-oestrus than from those of animals on the morning of dioestrus 1, raising the possibility that this hormone plays a physiological role in the regulation of prolactin secretion during the oestrous cycle.

Dependence of hyperpolarisation-activated cyclic nucleotide-gated channel activity on basal cyclic adenosine monophosphate production in spontaneously firing GH3 cells.

The hyperpolarisation-activated cyclic nucleotide-gated (HCN) channels play a distinct role in the control of membrane excitability in spontaneously active cardiac and neuronal cells. Here, we studied the expression and role of HCN channels in pacemaking activity, Ca(2+) signalling, and prolactin secretion in GH(3) immortalised pituitary cells. Reverse transcriptase-polymerase chain reaction analysis revealed the presence of mRNA transcripts for HCN2, HCN3 and HCN4 subunits in these cells. A hyperpolarisation of the membrane potential below - 60 mV elicited a slowly activating voltage-dependent inward current (I(h)) in the majority of tested cells, with a half-maximal activation voltage of -89.9 +/- 4.2 mV and with a time constant of 1.4 +/- 0.2 s at -120 mV. The bath application of 1 mM Cs(+), a commonly used inorganic blocker of I(h), and 100 microM ZD7288, a specific organic blocker of I(h), inhibited I(h) by 90 +/- 4.1% and 84.3 +/- 1.8%, respectively. Receptor- and nonreceptor-mediated activation of adenylyl and soluble guanylyl cyclase and the addition of a membrane permeable cyclic adenosine monophosphate (cAMP) analogue, 8-Br-cAMP, did not affect I(h). Inhibition of basal adenylyl cyclase activity, but not basal soluble guanylyl cyclase activity, led to a reduction in the peak amplitude and a leftward shift in the activation curve of I(h) by 23.7 mV. The inhibition of the current was reversed by stimulation of adenylyl cyclase with forskolin and by the addition of 8-Br-cAMP, but not 8-Br-cGMP. Application of Cs(+) had no significant effect on the resting membrane potential or electrical activity, whereas ZD7288 exhibited complex and I(h)-independent effects on spontaneous electrical activity, Ca(2+) signalling, and prolactin release. These results indicate that HCN channels in GH(3) cells are under tonic activation by basal level of cAMP and are not critical for spontaneous firing of action potentials.

Baclofen, a gamma-aminobutyric acid B agonist, modifies hormonal secretion in pituitary cells from infantile female rats.

Recent work from our laboratory has demonstrated that the activation of GABA B adenohypophyseal receptors by baclofen inhibits pituitary hormone secretion under basal (PRL) or stimulated conditions (PRL and LH) in adult female rats, suggesting a hypophyseal site of action in addition to the central site previously described. Since different patterns of hormone secretion are observed in infantile and adult rats, the purpose of the present study was to determine whether GABA B pituitary receptors were involved in endocrine responses at early stages of development. Pituitary cells of 12 day-old female rats were cultured in vitro and the effect of baclofen was determined in the presence or absence of stimulatory factors. Baclofen (1.10(-9), 1.10(-7) and 1.10(-5) M) did not alter basal LH or FSH secretion but significantly inhibited the LHRH induced gonadotropins release after 30 or 60 minutes of incubation (after 60 minutes of incubation LH (%): control: 100 +/- 5.6; BACL(1.10(-7)): 134.5 +/- 25.8; LHRH(1.10(-7)): 596.7 +/- 85.9; LHRH(1.10(-7))-BACL(1.10(-7)): 374.7 +/- 48.0; p<0.01. FSH (%): control: 100 +/- 6.5; BACL(1.10(-7): 103.7 +/- 6.5; LHRH(1.10(-7)): 283.9 +/- 29.3; LHRH(1.10(-7))-BACL(1.10(-7): 183.0 +/- 20.0; p<0.01). Baclofen did not significantly modify either basal or TRH-stimulated PRL or TSH secretion. These results show that baclofen has direct effects on the of adenohypophyseal cells of immature rats and such effects are different from those observed in adult rats, and depend on the stage of development of the neuroendocrine controls of each cellular type.