Reformulation of PULSAR for analysis of pulsatile LH secretion and a revised model of estrogen negative feedback in mice

The recent use of the tail-tip bleeding approach in mice has enabled researchers to generate detailed pulse and surge profiles of luteinizing hormone (LH) secretion in mice. However, the analysis of pulsatile LH secretion is piecemeal across the field with each laboratory using their own methodology. We have reformulated the once popular PULSAR algorithm of Merriam and Wachter to operate on contemporary computer systems and provide down-loadable and on-line pulse analysis platforms. As it is now possible to record the activity of the GnRH pulse generator in freely-behaving mice, we have been able to unambiguously define LH pulses in intact and gonadectomized male and female mice. These data sets were used to determine the appropriate PULSAR parameter sets for analysing pulsatile LH secretion in the mouse. This was then used to establish an accurate model of estrogen negative feedback in the mouse. Intact and ovariectomized mice given Silastic capsules containing 1, 2 and 4 µg 17-β-estradiol/20 gm body weight were tail-tip bled at 6-min intervals and the resultant LH profiles analysed with PULSAR. Only the 4 µg 17-β-estradiol capsule treatment was found to return LH pulse amplitude and frequency to that of intact diestrous mice. Ultra-sensitive mass spec analysis showed that the 4 µg 17-µ-estradiol capsule generated circulating estradiol levels equivalent to that of diestrous mice. It is hoped that the reformulation of PULSAR and generation of a realistic model of estrogen negative feedback will provide a platform for the more uniform assessment of pulsatile hormone secretion in mice.

Highly redundant neuropeptide volume co-transmission underlying episodic activation of the GnRH neuron dendron

The necessity and functional significance of neurotransmitter co-transmission remains unclear. The glutamatergic ‘KNDy’ neurons co-express kisspeptin, neurokinin B (NKB), and dynorphin and exhibit a highly stereotyped synchronized behavior that reads out to the gonadotropin-releasing hormone (GnRH) neuron dendrons to drive episodic hormone secretion. Using expansion microscopy, we show that KNDy neurons make abundant close, non-synaptic appositions with the GnRH neuron dendron. Electrophysiology and confocal GCaMP6 imaging demonstrated that, despite all three neuropeptides being released from KNDy terminals, only kisspeptin was able to activate the GnRH neuron dendron. Mice with a selective deletion of kisspeptin from KNDy neurons failed to exhibit pulsatile hormone secretion but maintained synchronized episodic KNDy neuron behavior that is thought to depend on recurrent NKB and dynorphin transmission. This indicates that KNDy neurons drive episodic hormone secretion through highly redundant neuropeptide co-transmission orchestrated by differential post-synaptic neuropeptide receptor expression at the GnRH neuron dendron and KNDy neuron.

Discrete-Time Mapping for an Impulsive Goodwin Oscillator with Three Delays

A popular biomathematics model of the Goodwin oscillator has been previously generalized to a more biologically plausible construct by introducing three time delays to portray the transport phenomena arising due to the spatial distribution of the model states. The present paper addresses a similar conversion of an impulsive version of the Goodwin oscillator that has found application in mathematical modeling, e.g. in endocrine systems with pulsatile hormone secretion. While the cascade structure of the linear continuous part pertinent to the Goodwin oscillator is preserved in the impulsive Goodwin oscillator, the static nonlinear feedback of the former is substituted with a pulse modulation mechanism thus resulting in hybrid dynamics of the closed-loop system. To facilitate the analysis of the mathematical model under investigation, a discrete mapping propagating the continuous state variables through the firing times of the impulsive feedback is derived. Due to the presence of multiple time delays in the considered model, previously developed mapping derivation approaches are not applicable here and a novel technique is proposed and applied. The mapping captures the dynamics of the original hybrid system and is instrumental in studying complex nonlinear phenomena arising in the impulsive Goodwin oscillator. A simulation example is presented to demonstrate the utility of the proposed approach in bifurcation analysis.

60 YEARS OF NEUROENDOCRINOLOGY: The posterior pituitary, from Geoffrey Harris to our present understanding

Geoffrey Harris pioneered our understanding of the posterior pituitary, mainly with experiments that involved the electrical stimulation of the supraoptico-hypophysial tract. In the present essay, we explain how his observations included clues to the pulsatile nature of the oxytocin signal – clues that were followed up by subsequent workers, including his students and their students. These studies ultimately led to our present understanding of the milk-ejection reflex and of the role of oxytocin in parturition. Discoveries of wide significance followed, including: the recognition of the importance of pulsatile hormone secretion; the recognition of the importance of stimulus-secretion coupling mechanisms in interpreting the patterned electrical activity of neurons; the physiological importance of peptide release in the brain; the recognition that peptide release comes substantially from dendrites and can be regulated independently of nerve terminal secretion; and the importance of dynamic morphological changes to neuronal function in the hypothalamus. All of these discoveries followed from the drive to understand the milk-ejection reflex. We also reflect on Harris's observations on vasopressin secretion, on the effects of stress, and on oxytocin secretion during sexual activity.