Gilbert R. Pitts

Mechanisms underlying episodic gonadotropin-releasing hormone secretion

The episodic secretion of gonadotropin-releasing hormone (GnRH) is crucial for fertility, but the cellular mechanisms and network properties generating GnRH pulses are not well understood. We will explore three primary aspects of this intermittent hormonal signal: the source of rhythm(s), the possible mechanisms comprising oscillator(s), and how GnRH neurons are synchronized to produce a pulse of hormone release into the pituitary portal blood. Current knowledge will be reviewed, and hypotheses and working models proposed for future studies.

Daily rhythmicity of large-conductance Ca2+-activated K+ currents in suprachiasmatic nucleus neurons

Neurons within the suprachiasmatic nucleus (SCN) comprise the master circadian pacemaker in mammals. These neurons exhibit circadian rhythms in spontaneous action potential frequency and in the transcription of core circadian clock genes, including Period1 (Per1). Targeted electrophysiological recordings from SCN neurons marked with a green fluorescent protein (GFP) reporter of Per1 gene transcription have previously indicated that K(+) currents are critically involved in the expression of neurophysiological rhythmicity. The present study examined the role of large conductance, Ca(2+)-activated K(+) channels (BK) in the daily rhythmicity of mouse SCN neurons. BK-mediated currents were examined in Per1::GFP neurons under voltage clamp using iberiotoxin, a specific BK channel blocker. BK current was a greater proportion of whole-cell outward currents during the night than during the day. Analysis of iberiotoxin difference currents also demonstrated that BK current amplitude and density were greater during the night and that the day/night difference in steady state amplitude was not due to altered inactivation. Single cell RT-PCR demonstrated the presence of the BK channel transcript, KCNMA1, in Per1-expressing neurons. In situ hybridization analysis further showed that KCNMA1 mRNA was rhythmically expressed in the SCN under light:dark (LD) conditions, peaking during the middle of the night phase. Acute inhibition of BK currents blunted the circadian rhythm SCN neuron spike frequency. These results establish that BK channel function is elevated at night, thus altering SCN neuron activity.

Vasoactive intestinal peptide stimulates prolactin mRNA expression in turkey pituitary cells: effects of dopaminergic drugs.

Abstract It is well documented that vasoactive intestinal peptide (VIP) is a prolactin (PRL)-releasing factor and that dopamine (DA) is an inhibitory neurotransmitter in avian species. However, the roles of VIP and DA in the regulation of PRL gene expression are unclear. In this study, primary anterior pituitary cells cultured from laying turkeys were utilized to investigate the influence of VIP and dopaminergic D1 and D2 receptors on PRL secretion, PRL mRNA, and PRL synthesis. Incubation of pituitary cells with VIP increased PRL secretion up to 3.5-fold within 3 hr. Prolactin mRNA was undetectable during the first 2 hr of pituitary cell treatment; thereafter, the PRL mRNA content response to VIP increased within 24-48 h (P < 0.05). Total PRL content (media + cellular) increased over time in the presence of VIP. The response of cells incubated in the presence of a dopaminergic D1 receptor agonist (SKF38393) was variable and inconclusive. However, cells incubated with a dopaminergic D2 receptor agonist (quin-pirole) inhibited VIP-induced PRL secretion (P < 0.05) and PRL mRNA levels (P < 0.05) in a dose-related fashion without effect on the basal levels of PRL release and PRL mRNA. These observations suggest that VIP, in addition to acting as a PRL-releasing peptide, also plays a role in the regulation of PRL gene expression. Moreover, the results of this study also indicate that a drug that can selectively stimulate dopamine D2 receptors can also regulate PRL secretion and PRL mRNA in turkey pituitary cells in culture. [P.S.E.B.M. 1996, Vol 212]

An Opioid Pathway in the Hypothalamus of the Turkey That Stimulates Prolactin Secretion

Circulating prolactin (PRL) levels increase when dynorphin is infused into the turkey brain. This study tested the hypothesis that centrally infused dynorphin requires an intact vasoactive intestinal peptide (VIP) system in order to stimulate turkey PRL secretion. It also investigated the roles of the dopaminergic and serotonergic systems in dynorphin-induced PRL release. Drugs were infused into the third ventricle of anesthetized laying turkeys via stereotaxically guided cannulae and circulating blood was assayed for changes in PRL levels. When a highly selective ĸ opioid receptor antagonist was given prior to dynorphin injection, the PRL response to dynorphin was almost totally blocked. The coinfusion of either a serotonin (5-HT) or a D1 dopamine (DA) receptor antagonist with dynorphin prevented the increase in PRL observed in birds when dynorphin was infused alone. On the other hand, the ĸ opioid receptor antagonist failed to prevent the 5-HT-induced release of PRL. In hens actively immunized against VIP, infused dynorphin was unable to increase plasma PRL levels and infused VIP gave a muted PRL rise, while large increases in PRL were seen in nonimmunized birds receiving the same infusions. These data show that: (1) dynorphin stimulates PRL secretion by activating ĸ opioid receptors in the avian hypothalamus, and (2) dynorphin, 5-HT, DA, and VIP stimulate avian PRL secretion via a common pathway expressing ĸ opioid, serotonergic, dopaminergic, and VIPergic receptors at synapses arranged serially in that functional order, with the VIPergic system as the final mediator (releasing factor).