Multiunit Electrical Activity Research Articles

New methods to investigate the GnRH pulse generator.

The exact neural construct underlying the dynamic secretion of gonadotrophin-releasing hormone (GnRH) has only recently been identified despite the detection of multiunit electrical activity volleys associated with pulsatile luteinising hormone (LH) secretion four decades ago. Since the discovery of kisspeptin/neurokinin B/dynorphin neurons in the mammalian hypothalamus, there has been much research into the role of this neuronal network in controlling the oscillatory secretion of gonadotrophin hormones. In this review, we provide an update of the progressive application of cutting-edge techniques combined with mathematical modelling by the neuroendocrine community, which are transforming the functional investigation of the GnRH pulse generator. Understanding the nature and function of the GnRH pulse generator can greatly inform a wide range of clinical studies investigating infertility treatments.

Left cardiac vagotomy rapidly reduces contralateral cardiac vagal electrical activity in anesthetized Göttingen minipigs

BackgroundThe impact of acute unilateral injury on spontaneous electrical activity in both vagus nerves at the heart level is poorly understood. We investigated the immediate neuroelectrical response after right or left cardiac vagal nerve transection (VNTx) by recording spiking activity of each heart vagus nerve (VN). MethodsFourteen male Göttingen minipigs underwent sternotomy. Multi-electrode cuffs were implanted below the cut level to record vagal electroneurographic signals during electrocardiographic and hemodynamic monitoring, before and immediately after cardiac VNTx (left: L-cut, n = 6; right: R-cut, n = 8). ResultsLeft cardiac VNTx significantly reduced multi-unit electrical activity (MUA) firing rate in the vagal stump (−30.7% vs pre-cut) and intact right VN (−21.8% vs pre-cut) at the heart level, without affecting heart rate, heart rate variability, or hemodynamics. In contrast, right cardiac VNTx did not acutely alter MUA in either VN but slightly increased (p < 0.022) the root mean square of successive RR interval differences (rMSSD), an index of parasympathetic outflow, without affecting hemodynamics. ConclusionsOur study reveals an early left-lateralized pattern in vagal spiking activity following unilateral cardiac vagotomy. These findings enhance understanding of the neuroelectrical response to vagal injury and provide insights into preserving vagal outflow after unilateral cardiac vagotomy. Importantly, monitoring spiking activity of the cardiac right VN may predict onset of left vagal pathway injury, which is detrimental to cardiac patients and can occur as a complication of catheter ablation for atrial fibrillation.

Electrophysiological Characterization of Cerebellar Responses during Exploration and Grooming Behaviors in a Rat Model of Parkinsonism.

Parkinson's disease is currently a global public health challenge due to the rapid growth of aging populations. To understand its pathophysiology is necessary to study the functional correlation between the basal ganglia (BG) and the cerebellum, which are involved in motor control. Herein, we explored multiunit electrical activity (MUA) in the cerebellum of rats with induced Parkinsonism as a result of lesions following bilateral placement of electrodes and passing of current in the ventrolateral striatum (VLS). In one control group, the electrodes descended without electrical current, and another group was left intact in VLS. MUA was recorded in Sim B and Crus II lobes, and in the dentate nucleus (DN) during the execution of exploration behaviors (horizontal and vertical) and grooming. The lesioned and sham groups showed a decrease in MUA amplitude in the Crus II lobe compared to the intact group in all recorded behaviors. However, Sim B and DN did not express differences. Both electrical and physical insults to the VLS induced Parkinsonism, which results in less MUA in Crus II during the execution of motor behaviors. Thus, this type of Parkinsonism is associated with a decrease in the amplitude of Crus II.

Neuroendocrine Mechanisms Regulating Reproductive Transition in the Ruminant Female: Is KNDy Required?

Abstract States of reproductive transition in female livestock represent important and economically significant periods of physiological change that have major impacts on the efficiency and success of breeding and production programs. These include, but are not limited to, the process of sexual maturation, postpartum resumption of ovarian cycles, and cyclical recrudescence in seasonal breeding species. Over the last four decades, our laboratory has focused primarily on determining the fundamental neuroendocrine mechanisms controlling states of reproductive transition in bovine and equine models, and applying principals learned to develop managerial approaches for positive intervention. The objectives of this presentation are to briefly review our understanding of external and internal signaling pathways that regulate two of these transitional states in the bovine female, postpartum resumption of ovulatory cycles and pubertal development. The focus will be to consider contrasts and commonalities of hypothalamic-pituitary signaling pathways that govern the temporal trajectories of these processes, with particular emphasis on those pathways that modulate the GnRH pulse generator. Notably (and until recently), the pulse generator has been a poorly defined “black box” with respect to its cellular origin and neurochemical character. Thus, the synchronous pattern of GnRH neuronal depolarization, multiunit electrical activity, and release of GnRH for the control of gonadotropins have been accepted as intrinsic features of GnRH neurons themselves. However, the identification of the neuropeptide kisspeptin in 1996, and more recently (2007) the description of a specialized population of kisspeptin-secreting neurons in the medial basal hypothalamus, termed KNDy neurons, have provided convincing evidence for an extrinsic source of pulse generation within the arcuate/infundibular region. How do KNDy neurons serve in this function, what is their role in transducing environmental, nutritional, and behavioral signals during transitional states, and what is the potential for neuropeptides of KNDy neuron origin to serve as pharmacological agents to control reproduction?

A121 SILENCING ENTERIC SENSORY NEURONS ABOLISHES THE EXCITATORY EFFECT OF SERTRALINE ON VAGAL AFFERENTS

Abstract Background Two thirds of vagal afferents supplying the small intestine terminate on intrinsic primary afferent neurons (IPANs) of the enteric nervous system via intraganglionic laminar endings (IGLEs) rather than the luminal epithelium (FASEB J, 28, 3064–3074, 2014). The IPAN to IGLE connection forms a functional intramural sensory synapse within the myenteric plexus. We have recently shown (Sci Rep, 9, 14290, 2019) that the antidepressant behavioural effects of selective serotonin reuptake inhibitors (SSRIs) are dependent on the vagus nerve and that intraluminal application of the SSRIs fluoxetine and sertraline increase vagal afferent firing rate in vitro. Aims We hypothesize that the vagal afferent response to intraluminal sertraline is mediated by IPAN to IGLE sensory signaling. Methods Mesenteric nerve recordings were performed using 2–3 cm jejunal segments and attached mesentery from 6–8 weeks old male Balb/c mice. Jejunal tissue was pinned out and dissected in a petri dish of Krebs to remove excess mesentery to isolate the mesenteric nerve bundle. Multi-unit electrical activity was recorded by patch-clamp electrode using an amplifier and signal converter by sucking onto the mesenteric nerve bundle with a glass micropipette. Baseline firing was recorded for 30 mins during luminal Krebs perfusion. The vagal afferent firing response to sertraline (10 µM) was measured in the absence or presence of 5 µM of the intermediate conductance calcium-dependent (IKCa) channel opener 5,6-dichloro-1-ethyl-1,3-dihydro-2H-benzimidazole-2-one (DCEBIO) to the serosa to selectively silence IPANs. Dataview software (J Neuroscience Methods, 185, 151, 2009) was used to isolate single unit firing post-hoc. Vagal fibre action potentials were identified by response to CCK. Results Intraluminal sertraline decreased the mean vagal interspike interval (increased vagal firing frequency) by 29% (p = 0.008) compared to Krebs control (N = 15 fibres). In contrast, addition of luminal sertraline in the presence of DCEBIO increased mean vagal interspike interval by 48% (p = 0.0103) compared to Krebs control (N = 12 fibres). Serosal addition of the N-type Ca2+ channel blocker w-Conotoxin GVIA simultaneously with luminal sertraline increased the mean vagal interspike interval by 12% (p = 0.0282) compared to Krebs control (N = 7 fibres). Conclusions Silencing of myenteric IPANs to vagus neurotransmission blocked excitatory response of vagal afferent fibres to intraluminal sertraline. Blocking myenteric neurotransmission reduced the vagus excitatory response to sertraline by more than 50%. These results suggest that the therapeutically necessary vagus nerve stimulation by sertraline involves activation of the myenteric intramural sensory synapse. Funding Agencies NSERC, OGS

Sensitivity and specificity of pulse detection using a new deconvolution method

Quantifying pulsatile secretion from serial hormone concentration measurements (deconvolution analysis) requires automated, objective, and accurate detection of pulse times to ensure valid estimation of secretion and elimination parameters. Lack of validated pulse identification constitutes a major deficiency in the deconvolution field, because individual pulse size and number reflect regulated processes that are critical for the function and response of secretory glands. To evaluate deconvolution pulse detection accuracy, four empirical models of true-positive markers of pituitary (LH) pulses were used. 1) Sprague-Dawley rats had recordings of hypothalamic arcuate nucleus multiunit electrical activity, 2) ovariectomized ewes underwent sampling of hypothalamo-pituitary gonadotropin-releasing hormone (GnRH pulses), 3) healthy young men were infused with trains of biosynthetic LH pulses after GnRH receptor blockade, and 4) computer simulations of pulsatile LH profiles were constructed. Outcomes comprised sensitivity, specificity, and receiver-operating characteristic curves. Sensitivity and specificity were 0.93 and 0.97, respectively, for combined empirical data in the rat, sheep, and human (n = 156 pulses) and 0.94 and 0.92, respectively, for computer simulations (n = 1,632 pulses). For simulated data, pulse-set selection by the Akaike information criterion yielded slightly higher sensitivity than by the Bayesian information criterion, and the reverse was true for specificity. False-positive errors occurred primarily at low-pulse amplitude, and false-negative errors occurred principally with close pulse proximity. Random variability (noise), sparse sampling, and rapid pulse frequency reduced pulse detection sensitivity more than specificity. We conclude that an objective automated pulse detection deconvolution procedure has high sensitivity and specificity, thus offering a platform for quantitative neuroendocrine analyses.

Effects of kisspeptin-10 on the electrophysiological manifestation of gonadotropin-releasing hormone pulse generator activity in the female rat.

Kisspeptins are extraordinarily potent in stimulating gonadotropic hormone secretion via an action on the hypothalamic GnRH neural system. Because the physiological frequency of the GnRH pulse generator is a critical component of the control system that governs reproductive processes, the aim of this study was to examine the effect of kisspeptin-10 on pulsatile LH secretion and on the electrophysiological manifestation of GnRH pulse generator activity to determine frequency modulatory effects. Adult Sprague Dawley rats were ovariectomized and chronically implanted with electrodes in the arcuate nucleus to record the characteristic increases in hypothalamic multiunit electrical activity volleys coincident with the initiation of each LH pulse measured in peripheral blood and/or indwelling cardiac catheters for the collection of blood samples (25 microl) every 5 min for 6-7 h for the measurement of LH. Intravenous infusion of kisspeptin-10 (7.5, 35, and 100 nmol) induced a dose-dependent increase in LH secretion. The stimulatory effect of kisspeptin-10 (100 nmol) on LH secretion was blocked by the GnRH antagonist cetrorelix, precluding a singular action on gonadotropes. Unexpectedly, however, the marked increase in LH release in response to kisspeptin-10 (100 nmol) administration was not accompanied by any change in multiunit electrical activity volley frequency. It seem unlikely, therefore, that kisspeptin-10 has an appreciable frequency modulatory effect on GnRH pulse generator activity in the female rat.

Simulation of Day-Length Encoding in the SCN: From Single-Cell to Tissue-Level Organization

The circadian pacemaker of the SCN is a heterogeneous structure containing many single-cell oscillators that display phase differences in gene expression and electrical activity rhythms. Thus far, it is unknown how single neurons contribute to the population signal measured from the SCN. The authors used single-unit electrical activity rhythms that have previously been recorded in SCN slices and investigated in simulation studies how changes in pattern shape and distribution of single neurons alter the ensemble activity rhythm of the SCN. The results were compared with recorded ensemble rhythms. The simulations show that single units should be distributed in phase to render the recorded multiunit waveform and that different distributions can account for the multiunit pattern of the SCN, including a bimodal distribution. Vice versa, the authors show that the single-unit distribution cannot be inferred from the ensemble pattern. Photoperiodic encoding by the SCN relies on changes in waveform of the neuronal output from the SCN and received special attention in this study's simulations. The authors show that a broadening or narrowing of the multiunit pattern can be based on changes in phase differences between neurons, as well as on changes in the circadian pattern of individual neurons. However, these mechanisms give rise to differences in the maximal discharge level of the multiunit pattern, leading to testable predictions to distinguish between the 2 mechanisms. If single units broaden their activity pattern in long days, the maximum frequency of the multiunit activity should increase, while an increase in phase difference between the single-unit activity rhythms should lead to a decrement in maximum frequency. The simulations also show that coding for day-length by an evening and morning oscillator is not self-evident and will only work under a limited set of conditions in which the distribution within each component and temporal distance between the components is taken into account. While the simulations were based on single-cell and multiunit electrical activity patterns, they are also relevant for understanding the relation between single-cell and population molecular expression profiles.

Targeted disruption of the mouse Mel(1b) melatonin receptor.

Two high-affinity, G protein-coupled melatonin receptor subtypes have been identified in mammals. Targeted disruption of the Mel(1a) melatonin receptor prevents some, but not all, responses to the hormone, suggesting functional redundancy among receptor subtypes (Liu et al., Neuron 19:91-102, 1997). In the present work, the mouse Mel(1b) melatonin receptor cDNA was isolated and characterized, and the gene has been disrupted. The cDNA encodes a receptor with high affinity for melatonin and a pharmacological profile consistent with its assignment as encoding a melatonin receptor. Mice with targeted disruption of the Mel(1b) receptor have no obvious circadian phenotype. Melatonin suppressed multiunit electrical activity in the suprachiasmatic nucleus (SCN) in Mel(1b) receptor-deficient mice as effectively as in wild-type controls. The neuropeptide, pituitary adenylyl cyclase activating peptide, increases the level of phosphorylated cyclic AMP response element binding protein (CREB) in SCN slices, and melatonin reduces this effect. The Mel(1a) receptor subtype mediates this inhibitory response at moderate ligand concentrations (1 nM). A residual response apparent in Mel(1a) receptor-deficient C3H mice at higher melatonin concentrations (100 nM) is absent in Mel(1a)-Mel(1b) double-mutant mice, indicating that the Mel(1b) receptor mediates this effect of melatonin. These data indicate that there is a limited functional redundancy between the receptor subtypes in the SCN. Mice with targeted disruption of melatonin receptor subtypes will allow molecular dissection of other melatonin receptor-mediated responses.

Phytoestrogens and gonadotropin-releasing hormone pulse generator activity and pituitary luteinizing hormone release in the rat.

Phytoestrogens can produce inhibitory effects on gonadotropin secretion in both animals and humans. The aims of this study were 2-fold: 1) to determine in vivo whether genistein and coumestrol act on the GnRH pulse generator to suppress hypothalamic multiunit electrical activity volleys and associated LH pulses and/or on the pituitary to suppress the LH response to GnRH; and 2) to examine the effect of these phytoestrogens on GnRH-induced pituitary LH release in vitro and to determine whether estrogen receptors are involved. Wistar rats were ovariectomized and chronically implanted with recording electrodes and/or indwelling cardiac catheters, and blood samples were taken every 5 min for 7--11 h. Intravenous infusion of coumestrol (1.6-mg bolus followed by 2.4 mg/h for 8.5 h) resulted in a profound inhibition of pulsatile LH secretion, a 50% reduction in the frequency of hypothalamic multiunit electrical activity volleys, and a complete suppression of the LH response to exogenous GnRH. In contrast, both genistein (1.6-mg bolus followed by 2.4 mg/h for 8.5 h) and vehicle were without effect on pulsatile LH secretion. Coumestrol (10(-5) M; over 2 or 4 h) suppressed GnRH-induced pituitary LH release in vitro, an effect blocked by the antiestrogen ICI 182,780. It is concluded that coumestrol acts centrally to reduce the frequency of the hypothalamic GnRH pulse generator. In addition, the inhibitory effects of coumestrol on LH pulses occur at the level of the pituitary by reducing responsiveness to GnRH via an estrogen receptor-mediated process.

Estradiol increases multiunit electrical activity in the A15 area of ewes exposed to inhibitory photoperiods.

Seasonal anestrus in ewes results from an increase in response to the negative feedback action of estradiol (E(2)). This increase in the inhibitory effects of E(2) is controlled by photoperiod and appears to be mediated, in part, by dopaminergic neurons in the retrochiasmatic area of the hypothalamus (A15 group). This study was designed to test the hypothesis that E(2) increases multiunit electrical activity (MUA) in the A15 during inhibitory long days. MUA was monitored in the retrochiasmatic area of 14 ovariectomized ewes from 4 h before to 24 h after insertion of an E(2)-containing implant subcutaneously. In six of these ewes, MUA activity was also monitored before and after insertion of blank implants. Three of the 14 ewes were excluded from analysis because E(2) failed to inhibit LH. When MUA was recorded within the A15, E(2) produced a gradual increase in MUA that was sustained for 24 h. Blank implants failed to increase MUA in the A15 area, and E(2) did not alter MUA if recording electrodes were outside the A15. These data demonstrate that E(2) increases MUA in the A15 region of ewes and are consistent with the hypothesis that these neurons mediate E(2) negative feedback during long photoperiods.

The Wisdom of the Body Revisited.

Being selected a Cannon lecturer by the American Physiological Society is a singular honor under any circumstances, but a particularly poignant one for me because of my origins in Cannon’s old Department at the Harvard Medical School. In fact, when I began my career there as an instructor 45 years

On the role of gonadotropin-releasing hormone (GnRH) in the operation of the GnRH pulse generator in the rhesus monkey.

The pulsatile secretion of luteinizing hormone (LH), occasioned by the pulsatile release of gonadotropin-releasing hormone (GnRH), is closely associated with concurrent increases in multiunit electrical activity in the mediobasal hypothalamus (MUA volleys), the electrophysiological correlates of GnRH pulse generator activity. These volleys represent a highly synchronized increase in firing frequency of individual neurons. The origin of these rhythmic oscillations in unit activity and the mechanisms responsible for their synchronization are unknown. The purpose of the present study was to examine the role, if any, of GnRH in the functioning of the GnRH pulse generator in rhesus monkeys. Ovariectomized animals bearing recording electrodes chronically implanted in the mediobasal hypothalamus and fitted with intracerebroventricular (ICV) cannulae in the lateral ventricle and with indwelling cardiac catheters were studied. LH was measured in venous blood withdrawn from the cardiac catheters every 10 min while hypothalamic electrical activity was monitored continuously. In Experiment 1, following a 3- to 4-hour control period, GnRH was infused ICV at a rate of 300 ng/kg body weight (BW)/h over 4-5 h. In Experiment 2, antide, a long-acting GnRH antagonist, was injected ICV in a dose of 105 microg/kg BW after a control period of 3-4 h. Additional control experiments were performed in each animal using vehicle alone. Neither GnRH nor antide affected the frequency of MUA volleys and attendant LH pulses despite significant alterations in LH secretion. These results suggest that, in the rhesus monkey, GnRH may not be involved in the operation of the GnRH pulse generator.

Hypothalamic multiunit activity and pulsatile luteinizing hormone release in the castrated male rat

Using chronically implanted microelectrodes, multiunit electrical activity (MUA) was recorded from the arcuate nucleus of freely moving gonadectomized male rats. Intermittent increases in MUA activity (MUA volleys) closely associated with luteinizing hormone pulses measured in the peripheral circulation were observed, which confirms that this experimental approach can be used for monitoring the activity of the gonadotropin-releasing hormone-associated hypothalamic pulse generator in the male rat. The mean MUA volley frequency was 22.2 min (range 13-38 min), whereas the mean MUA volley duration was 2.7 +/- 0.8 min (standard deviation). In addition to a large inter-individual variability. MUA volley intervals also showed an important intra-individual variability. This observation suggests that, beside the mean frequency of pulse generator activation, the degree of variability in gonadotropin-releasing hormone-associated pulse generator activity might be an additional relevant parameter in the characterization of the reproductive function in the male rat.

Electrophysiological approaches to gonadotrophin releasing hormone pulse generator activity in the rhesus monkey.

Precipitous increases in multi-unit electrical activity (MUA volleys) can be recorded from the mediobasal hypothalamus of the rhesus monkey that are invariably synchronous with the initiation of luteinizing hormone (LH) pulses and thus serve as markers of gonadotrophin releasing hormone (GnRH) pulse generator activity. Continuous radiotelemetric monitoring of this activity throughout the menstrual cycle confirmed a lower pulse generator frequency during the luteal phase than in the follicular phase. Unexpectedly, however, just before the initiation of the LH surge an abrupt reduction in frequency was observed, followed, on occasion, by a total arrest of GnRH pulse generator activity for a day or two. This monitoring technique also revealed a reduction of pulse generator frequency at night. This appeared to be an acute phenomenon observable in the first MUA volleys after the lights are turned off, as is the increase in frequency when the lights are turned on in the morning suggesting a direct action of light, a conclusion that was verified using lighting periods other than the customary ones. This action of light, however, is superimposed upon a diurnal rhythm, as shown by a reduction in pulse generator frequency during the subjective night when the monkeys are kept in constant light or darkness. The duration of the MUA volley in the intact monkey is 1-3 min, compared with some 20 min in the ovariectomized female. While oestradiol reduces the volley duration in the ovariectomized female to that of intact monkeys within 6 h, it takes some 6 weeks following ovariectomy for the duration characteristic of castrates to be re-established.(ABSTRACT TRUNCATED AT 250 WORDS)

Single unit components of the hypothalamic multiunit electrical activity associated with the central signal generator that directs the pulsatile secretion of gonadotropic hormones.

Vertebrate reproduction is dependent on the operation of a central signal generator that directs the episodic release of gonadotropin-releasing hormone, a neuropeptide that stimulates secretion of the pituitary gonadotropic hormones and, thereby, controls gonadal function. The electrophysiological correlates of this pulse generator are characterized by abrupt increases in hypothalamic multiunit electrical activity (MUA volleys) invariably associated with the initiation of secretory episodes of luteinizing hormone. Using cluster analysis, we extracted single units from the multiunit signals recorded from the mediobasal hypothalamus of four intact and four ovariectomized rhesus monkeys. Of the 40 individual units identified in this manner, 24 increased their frequency with the MUA volleys. The onset and termination of these single-unit bursts occurred coincidently with those of the MUA volleys in both intact and ovariectomized animals, indicating that the longer duration of the MUA volleys characteristic of the gonadectomized animals was due not to the sequential activation of different units but to the longer bursts of the individual cells. Four other units showed decreases in firing rate during the MUA volleys, while the frequency of the remainder did not change. All the examined units were active during the intervals between the volleys of electrical activity. The results indicate that the MUA volleys associated with the activity of the gonadotropin-releasing hormone pulse generator represent the simultaneous increase in firing rate of some individual hypothalamic neurons and the decrease in the frequency of others.

Ambient light modifies gonadotropin-releasing hormone pulse generator frequency in the rhesus monkey.

In the course of previous studies using continuous monitoring of the electrophysiological correlates of GnRH pulse generator activity, characterized by episodic increases in hypothalamic multiunit electrical activity (MUA volley), it was noticed that the nocturnal slowing of pulse generator frequency was an acute phenomenon observable in the first MUA volleys after the lights were turned off, as was the increase in frequency when the lights were turned on in the morning. This suggested that the reduction in pulse generator frequency at night may not be the consequence of an intrinsic diurnal rhythm, but an effect of light per se. Indeed, as reported herein, such an effect was observed when the lights were turned on or off at times other than the normal illumination period (normal light schedule, lights on from 0700-1900 h). That this was not simply a response to arousal was shown by awakening the animals with loud recorded noises in total darkness at the same unaccustomed times without a resulting change in frequency. This suggests that the effect of light is specific, perhaps mediated by the retino-hypothalamic tract. This direct action of light, however, is superimposed upon a diurnal rhythm, as shown by a reduction in pulse generator frequency during the subjective night when the monkeys were kept in constant light or constant darkness.

Ovarian control of gonadotropin hormone-releasing hormone pulse generator activity in the rhesus monkey: duration of the associated hypothalamic signal.

The activity of the GnRH pulse generator in the rhesus monkey is associated with abrupt increases in multiunit electrical activity (MUA) volleys recorded from the mediobasal hypothalamus that precede each pulse of LH in the peripheral circulation. In long-term ovariectomized animals the duration of these MUA volleys is 10-25 min and consists of a brief initial 'overshoot' followed by a plateau phase that ends in a rapid decline to baseline activity. In intact monkeys, however, the MUA volley lasts only 1-3 min, a duration equivalent to the overshoot in ovariectomized animals. In addition, the maximal frequency of neuronal activity during each MUA volley is reduced in normal animals when compared to castrates. As shown in earlier studies, estradiol given to ovariectomized monkeys causes a reduction in the duration of MUA volleys to that characteristic of intact animals within 3-5 h. In contrast to this acute effect of estradiol, the increase in MUA volley duration following ovariectomy is a gradual phenomenon, 4-6 weeks being required to achieve the MUA volley duration observed in long-term ovariectomized monkeys. A similar slow time course was observed for the increase in maximal neuronal frequency during each MUA volley. This protracted effect of ovariectomy on MUA volley duration and firing rate may be the consequence of hypothalamic remodelling but this consideration must be tempered by the observation that estradiol reverses these phenomena within hours.

Effects of naloxone on estrogen-induced changes in hypothalamic gonadotropin-releasing hormone pulse generator activity in the rhesus monkey.

In the ovariectomized rhesus monkey, estradiol (E2) markedly reduces the frequency of the GnRH pulse generator as monitored by LH pulse frequency and the concurrent changes in hypothalamic electrical activity, an action mimicked by morphine. In addition, the duration of the increments in multiunit electrical activity (MUA volleys) that precede each LH pulse is decreased by estrogen administration, an action also shared by morphine. The role of endogenous opioids in these actions of E2 was investigated in 8 ovariectomized animals restrained in primate chairs. They were fitted with indwelling cardiac catheters and with bilateral arrays of recording electrodes chronically implanted in the mediobasal hypothalamus. Physiological serum E2 levels achieved by subcutaneous implantation of E2-containing Silastic capsules increased MUA volley interval from 50.8 +/- (SEM) 1.6 min in the control period to 81.1 +/- 6.2 min following E2. Mean MUA volley duration decreased from 21.9 +/- 1.0 to 13.0 +/- 0.7 min. The placement of empty Silastic capsules had no effect on MUA volley duration or interval. Naloxone administration (2.5 mg bolus followed by a 1 mg/h infusion lasting 4-8 h) completely (n = 4) or partially (n = 2) blocked the effects of E2 on MUA volley interval in 6 of the 8 monkeys, and was without effect in the remainder. In contrast, however, naloxone had little or no effect on the action of E2 on MUA volley duration, (13.0 +/- 0.7 vs. 14.0 +/- 0.9 min). These findings suggest that the inhibitory action of E2 on GnRH pulse generator frequency, like that of all other gonadal steroids studied to date, may be mediated by endogenous opioids.(ABSTRACT TRUNCATED AT 250 WORDS)

Melatonin modifies the spontaneous multiunit activity recorded in several brain nuclei of freely behaving rats

Melatonin, a pineal hormone, released photoperiodically, was administered systemically in rats, previously implanted with semimicroelectrodes into six different brain structures. The multiunit electrical activity of these structures was recorded for 10 min before and 60 min after melatonin administration in unanesthetized, freely moving rats. Different melatonin doses (100, 200, 500, and 1000 μg/kg) produced changes in the electrical activity of all tested structures. However, amygdala, rostral hypothalamus and mesencephalic reticular formation showed the most important changes. The main effect induced by melatonin was a dose-related decrease of the spontaneous electrical activity. The significance of these effects is discussed within the context of the behavioral and endocrinological effects of melatonin.