Hypothalamic Multiunit Activity Research Articles

The insulin hypoglycemia-induced inhibition of gonadotropin-releasing hormone pulse generator activity in the rhesus monkey: roles of vasopressin and corticotropin-releasing factor.

Insulin-induced hypoglycemia (IIH) profoundly inhibits the activity of the hypothalamic GnRH pulse generator. The aim of this study was to determine the role of vasopressin and CRF in this response. Ovariectomized rhesus monkeys with chronically implanted recording electrodes in the mediobasal hypothalamus and with intracerebroventricular (icv) cannulas in the lateral ventricle were placed in primate chairs, and blood samples were taken every 10 min. Pulse generator activity was monitored electrophysiologically by detecting characteristic increases in hypothalamic multi-unit activity (MUA volleys) and by attendant LH pulses measured in peripheral blood. Arginine vasopressin (AVP) infused via the i.c.v. cannula (50 micrograms/60 microliters.h) in eight monkeys failed to decrease pulse generator activity, as measured by the frequency of MUA volleys, but decreased mean serum LH concentrations (P < 0.001) while increasing serum cortisol levels (P < 0.02). Central administration of an AVP antagonist ([deamino-Pen1, O-Me-Tyr2-Arg8] vasopressin) in four monkeys at a rate (180 micrograms/60 microliters.h) that had previously been found to block the aforementioned effects of coadministered AVP failed to prevent the IIH-induced inhibition of GnRH pulse generator activity and LH secretion in the same animals. On the other hand, a CRF antagonist, [D-Phe12,Nle21,38,C alpha MeLeu37] rat CRF-(12-41), infused i.c.v. at a rate of 500 micrograms/60 microliters.h in four monkeys, delayed the inhibition of pulse generator frequency in response to IIH. These results suggest that AVP does not mediate the hypoglycemia-induced inhibition of GnRH pulse generator frequency in the rhesus monkey, but that CRF may be involved in this response.

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.

Estradiol and the inhibition of hypothalamic gonadotropin-releasing hormone pulse generator activity in the rhesus monkey.

In mammals, gonadal function is controlled by a hypothalamic signal generator that directs the pulsatile release of gonadotropin-releasing hormone (GnRH) and the consequent pulsatile secretion of luteinizing hormone. In female rhesus monkeys, the electrophysiological correlates of GnRH pulse generator activity are abrupt, rhythmic increases in hypothalamic multiunit activity (MUA volleys), which represent the simultaneous increase in firing rate of individual neurons. MUA volleys are arrested by estradiol, either spontaneously at midcycle or after the administration of the steroid. Multiunit recordings, however, provide only a measure of total neuronal activity, leaving the behavior of the individual cells obscure. This study was conducted to determine the mode of action of estradiol at the level of single neurons associated with the GnRH pulse generator. Twenty-three such single units were identified by cluster analysis of multiunit recordings obtained from a total of six electrodes implanted in the mediobasal hypothalamus of three ovariectomized rhesus monkeys, and their activity was monitored before and after estradiol administration. The bursting of all 23 units was arrested within 4 h of estradiol administration although their baseline activity was maintained. The bursts of most units reappeared at the same time as the MUA volleys, the recovery of some was delayed, and one remained inhibited for the duration of the study (43 days). The results indicate that estradiol does not desynchronize the bursting of single units associated with the GnRH pulse generator but that it inhibits this phenomenon. The site and mechanism of action of estradiol in this regard remain to be determined.

Acute Estradiol Modulation of Electrical Activity of the LHRH Pulse Generator in the Ovariectomized Rat: Restoration by Naloxone

Whether estrogen has the brain as a site of its negative feedback action was investigated by checking the acute effect of estradiol-17 beta (E2) on the electrical activity of the luteinizing hormone-releasing hormone (LHRH) pulse generator in ovariectomized rats fitted with chronically implanted electrode arrays in the medial basal hypothalamus. Before subcutaneous E2 implantation, the hypothalamic multiunit activity (MUA) exhibited, at an average frequency of 2.43/h, characteristic increases (volleys), each of which was associated with the initiation of an LH pulse. Within 2 h after E2 implantation, the frequency of MUA volleys was reduced significantly, probably associated with decreases in the frequency of LH pulses. The decrease in the amplitude of LH pulses occurred later than 2 h, but this effect was considered not to be mediated by the brain since the duration of MUA volleys was not changed. Since the mean serum LH concentration started to decrease within 2 h after E2 implantation, it was concluded that the acute inhibitory E2 effect on LH release is mediated by the brain. On the day after E2 implantation, intravenous infusion of naloxone (2 mg/kg/h) promptly elicited intermittent MUA volleys each of which was associated with an LH pulse. This suggests that operation of the LHRH pulse generator in the ovariectomized condition is restrained by the action of E2 with the mediation of endogenous opioid peptide neurons.

Cessation of the electrical activity of gonadotropin-releasing hormone pulse generator during the steroid-induced surge of luteinizing hormone in the rat.

Neuronal activity in the mediobasal hypothalamus was recorded during the steroid-induced surge of luteinizing hormone (LH) in ovariectomized female rats by means of multiunit activity (MUA) recording techniques. The day of subcutaneous implantation of estradiol-containing silastic tubing, which was done at 12.00 h, was designated as day 1, and progesterone (1 mg) dissolved in sesame oil was injected at 12.00 h on day 5. The hypothalamic MUA and serum LH level were monitored between 10.00 and 20.00 h on days 1, 4 and 5. All 5 animals examined showed characteristic increases (volleys) in MUA associated with pulsatile LH secretion. After implantation of estradiol on day 1, gradual and significant decreases in the serum LH level and in MUA volley frequency were observed. In the afternoon of both days 4 and 5, an LH surge occurred, though it was much greater on day 5 than day 4 due to progesterone injection on day 5. However, neither MUA volleys nor considerable changes in baseline MUA were discernible throughout the recording period on days 4 and 5. More than 5 days after the removal of estradiol-containing tubing, MUA volleys were found to reappear in all the animals. These results suggest that estradiol has an inhibitory effect on gonadotropin-releasing hormone (GnRH) pulse generator activity, and that the GnRH pulse generator is not involved in inducing the LH surge in the rat.

Luteinizing hormone (LH) pulsatility during the oestradiol- and oestradiol/progesterone-induced LH surge in the human female

Positive feedback reactions were induced in five female volunteers with regular menstrual cycles on the pituitary secretion of luteinizing hormone (LH) during the follicular phase of the cycle by the i.m. administration of oestradiol benzoate ('oestradiol study'), or by oestradiol benzoate followed by progesterone ('oestradiol/progesterone study'). Prior to the administration of the steroids (basal profiles) and during the LH surges following the administration of the steroids (stimulation profiles), blood was drawn at 10-min intervals and the LH pulsatility assessed. The LH pulses occurred at hourly intervals in basal and stimulation profiles, which concurs with data obtained during the normal proliferative phase and the spontaneous mid-cycle surge of the cycle in both the human and the rhesus monkey. Our findings are, however, in sharp contrast with those obtained in the rhesus monkey by the determination of hypothalamic multi-unit activity.

Effects of gamma-aminobutyric acid-A receptor antagonist, bicuculline, on the electrical activity of luteinizing hormone-releasing hormone pulse generator in the ovariectomized rat.

The role of GABA neurons in the control of pulsatile release of LHRH was investigated by checking the effect of the GABAA receptor agonist, muscimol, and antagonist, bicuculline, on the electrical activity of the luteinizing hormone-releasing hormone (LHRH) pulse generator in the ovariectomized rat fitted with chronically implanted electrode arrays in the medial basal hypothalamus. In untreated control animals, the hypothalamic multiunit activity (MUA) exhibited, at an average of 20.5-min intervals, characteristic increases (volleys), each of which was associated with the initiation of an LH pulse. A bolus i.v. injection of muscimol (2 mg/kg) significantly increased the interval between MUA volleys and LH pulses without affecting the pulse amplitude. Continuous i.v. infusion of saline increased the interval between MUA volleys to an average of 24.1 min without affecting the LH pulse amplitudes. Bicuculline infusion (10 mg/kg/h) altered neither the interval between MUA volleys nor the pulsatile release of LH. This was further checked in the condition where presynaptic inhibition by opioid peptides on the noradrenergic system was presumably decreased by naloxone. Naloxone infusion (0.5 or 0.7 mg/kg/h) caused the MUA volleys to occur markedly frequently, an average of 13.8-min intervals. The interval during combined infusion of bicuculline with naloxone was an average of 16.2 min, suggesting that bicuculline could not decrease the interval that was set by naloxone. The results show that, although exogenous GABAA receptor agonist is capable of inhibiting the activity of LHRH pulse generator, the reduction in the endogenous GABAA receptor activity does not cause a significant effect, suggesting a minor role of inhibitory GABA neurons in the control of pulsatile release of LHRH. Further, together with the well-known fact that activation of GABAA receptor hyperpolarizes neurons postsynaptically, it is assumed that the GABAergic system, unlike the opioidergic one, is not involved in the presynaptic inhibition of the adrenergic receptor system which is probably implicated in the frequency control of LHRH pulse generator.

Radiotelemetric Monitoring of Hypothalamic Gonadotropin-Releasing Hormone Pulse Generator Activity Throughout the Menstrual Cycle of the Rhesus Monkey*

Continuous monitoring of the electrophysiological manifestations of GnRH pulse generator activity was achieved by radiotelemetry throughout the menstrual cycles of unrestrained rhesus monkeys. The characteristic increases in hypothalamic multiunit activity (MUA volleys) associated with each LH pulse measured in the peripheral circulation were of lower frequency during the luteal phase than in the follicular phase of the cycle. Multiunit activity volley frequency increased as functional luteolysis progressed and achieved maxima of approximately one volley per hour within the first few days of the follicular phase. Unexpectedly, a dramatic decline in pulse generator frequency was observed coincidentally with the initiation of the preovulatory LH surge. Evidence is presented to support the conclusion that this deceleration of pulse generator activity is the consequence of the preovulatory rise in plasma estrogen concentration. As reported in women, a significant reduction in GnRH pulse generator frequency was observed at night during the follicular phase, but not during the luteal phase, of the menstrual cycle.

In VivoBiological Validation and Biophysical Modeling of the Sensitivity and Positive Accuracy of Endocrine Peak Detection. II. The Follicle-Stimulating Hormone Pulse Signal*

Despite interest in the in vivo control of gonadotropin release, valid assessment of the physiological regulation of the pulsatile secretion of the gonadotropin FSH has been hampered by the uncertain validity and reliability of available FSH peak detection algorithms. Difficulties in identifying FSH peaks accurately are believed to arise in part because of the slow metabolic clearance of this glycoprotein hormone. Here, we have used two complementary strategies to test the validity of FSH pulse detection. First, by means of a computer-assisted mathematical model for simulating episodic hormone secretion, we evaluated the effects of various putative FSH secretory pulse amplitudes and half-lives on the sensitivity and positive accuracy of peak detection. Secondly, we used an in vivo primate animal model, in which presumptively true FSH pulses were evaluated independently by continuous electrophysiological monitoring of mediobasal hypothalamic multiunit activity. These two approaches allowed us to define optimal pulse analysis parameters that yield maximal sensitivity and positive accuracy for detecting FSH peaks in synthetic and biological time series. We found (as predicted intuitively) that increasing half-times of hormone disappearance decrease both the sensitivity and positive accuracy of peak detection for any given peak detection thresholds and hormone secretory amplitudes. However, adequately sampled episodic FSH time series could be analyzed for FSH pulsatility by an appropriately constrained, objective computerized algorithm with reasonable (less than 10-15%) false negative and false positive errors, such that resultant sensitivity and positive accuracy exceed 85-90%. Of interest, computer simulations and the in vivo animal model exhibited similar discriminative capabilities. We conclude that increasing half-times of hormone (e.g. FSH) removal do impair hormone peak detection sensitivity and positive accuracy. Nevertheless, gonadotropin time series can be analyzed for FSH pulsatility in a valid manner with adequately constrained false negative and false positive error rates.

Interactions between the Noradrenergic and Opioid Peptidergic Systems in Controlling the Electrical Activity of Luteinizing Hormone-Releasing Hormone Pulse Generator in Ovariectomized Rats

The effects of noradrenergic and opioid peptidergic receptor blockade, either alone or in combination, on the electrical activity of luteinizing hormone-releasing hormone (LHRH) pulse generator were studied in ovariectomized rats fitted with chronically implanted electrode arrays in the medial basal hypothalamus. Both alpha- and beta-adrenergic receptor antagonists, i.e. phenoxybenzamine (5 mg/kg i.v.) and propranolol (5 mg/kg i.v.), respectively, significantly increased the intervals between characteristic increases (volleys) in hypothalamic multiunit activity (MUA), which were associated with the initiation of LH pulses. In contrast to this, an opioid receptor antagonist naloxone (2 mg/kg i.v.) significantly decreased the intervals between the MUA volleys. Naloxone given after the injection of propranolol induced MUA volleys with a latency of a few minutes. However, when given after the injection of phenoxybenzamine, naloxone failed to induce such immediate MUA volleys. These changes in the intervals between the MUA volleys were faithfully reflected by the pulsatile LH secretion. These results suggest that norepinephrine facilitates LHRH pulse generator activity through both alpha- and beta-adrenergic-receptors, and that the action of opioid peptides on it requires an alpha-adrenergic receptor-mediated mechanism.

Hypothalamic multiunit activity and LH secretion in conscious sheep.

The relationship between the pulsatile secretion of luteinizing hormone (LH) and multiunit activity (MUA) in the median eminence and retrochiasmatic area (RCh-ME) of the hypothalamus was investigated within 13 conscious, ovariectomized ewes during the anoestrous season. Eight of the ewes had been treated with oestradiol to reduce their LH pulse frequency. To test whether the activity had been recorded from sites involved in the control of GnRH release, we electrically stimulated the sites studied with the recording electrode and used the LH responses to classify the animals retrospectively for analysis of the MUA data. Following stimulation, LH secretion was either stimulated (Group STIM, n = 5 ewes), inhibited (Group INHIB, n = 4) or showed no response (Group NR, n = 4). Statistical analysis of the MUA data revealed that the onset of LH pulses was associated with an increase in cell activity in STIM ewes and a decrease in activity in both Group INHIB and Group NR ewes. Histological examination revealed that the electrodes were located near the midline in Group STIM ewes, but 1.25 or 2 mm lateral of the midline in Groups NR and INHIB, respectively. We concluded that the MUA in the RCh-ME probably reflects the activity of cells with fibres or terminals involved in the control of GnRH release. Activity which increased at the onset of LH pulses was detected in medial areas of the RCh-ME and may reflect the activity of GnRH neurones. The activity which decreased at the onset of LH pulses in lateral areas of the RCh-ME suggests that both stimulatory and inhibitory inputs may be involved in the release of GnRH pulses.

Hypothalamic multiunit activity correlates of the adrenocortical response to stress

The corticotropic response to stress was studied by means of multiple unit activity (MUA) recording from the adrenocorticotropic region of the hypothalamus, and plasma corticosterone (B) determination. MUA was permanently obtained and B was measured at 2, then 5 and 10 min intervals before and after neurogenic (electrical shocks) or systemic (ether inhalation) stress was applied. Experiments were made on steady unanesthetized, unrestrained thalamic pigeons. Post-stress alterations of MUA and B were closely in parallel, exhibiting a rapid and sustained increase in firing rate and shifting by 5–10 min, in corticosteronemia. Three successive and progressively decreasing peaks of MUA and B could be observed. Basal resting values were restored by approximately 90 min. Adrenocorticotropic responses to stress appear to be modulated through neural thalamic and/or rhombencephalic mechanisms.

Effects of gonadal hormones on the hypothalamic multi-unit response to glucose and saline in the rat.

The role of gonadal hormones in modulating the hypothalamic multi-unit activity (MUA) response to glucose and saline was studied in normal and gonadectomized male and female rats. In intact male rats, unlike intact females, all hypothalamic placements showed either an increase or a decrease in MUA following an injection of glucose. Gonadectomy alone did not produce a significant change in the pattern of hypothalamic response to glucose. Daily injections of estradiol-17beta resulted in a significant increase in the number of hypothalamic sites that did not show a glucose-induced change in MUA. In general, the effects on saline tended to parallel those on glucose, although the changes were not as extensive. The results are discussed of estradiol modulation of body weight (b.w.) and of the hypothalamic responsiveness to various stimuli, including glucosse and saline injections.