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Biomedical subjects

E Knobil

Publications and source records attributed to E Knobil.

At least 37 records · Page 2Linked to original sources

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.

Animals↗

Hypoglycemic 'stress' and gonadotropin-releasing hormone pulse generator activity in the rhesus monkey: role of the ovary.

Observations of long standing have suggested that the 'stress' of chair restraint inhibits the GnRH pulse generator in normal female monkeys while this phenomenon is rarely observed in ovariectomized animals. The role of the ovary in the response of the GnRH pulse generator to the stress of insulin hypoglycemia was investigated in both intact and ovariectomized rhesus monkeys. Following an overnight fast the animals, previously habituated to restraint, were placed in primate chairs and GnRH pulse generator activity monitored electrophysiologically. Insulin-induced reductions in mean blood glucose concentrations of 10-40% of control values interrupted pulse generator activity in intact monkeys but were without effect in ovariectomized animals. With larger reductions in blood glucose, pulse generator activity was interrupted in both groups but the inhibition was twice as long in intact than in ovariectomized animals. The reduced responsiveness of ovariectomized animals to insulin hypoglycemia was significantly reversed by estradiol replacement. Naloxone administration did not prevent the hypoglycemia-induced inhibition of pulse generator activity in either intact or ovariectomized rhesus monkeys. It is concluded that hypoglycemic 'stress' inhibits the GnRH pulse generator by a nonopioidergic mechanism and that ovarian products, most probably estradiol, exacerbate this effect.

Animals↗

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.

Animals↗

Duration of phasic electrical activity of the hypothalamic gonadotropin-releasing hormone pulse generator and dynamics of luteinizing hormone pulses in the rhesus monkey.

The secretion of luteinizing hormone (LH) by the pituitary gland is a pulsatile phenomenon. In the rhesus monkey, each pulse of LH in the peripheral circulation is associated with a characteristic increase in multiunit electrical activity (MUA) recorded from the medial basal hypothalamus. These "volleys" of electrical activity initiate the release of gonadotropin-releasing hormone (GnRH) into the pituitary portal circulation from the terminals of neurosecretory cells. Their duration varies from 1-3 min in normal, adult intact females to 10-25 min in long-term ovariectomized monkeys. A variety of pharmacological interventions also modify volley duration. The purpose of this investigation was to determine the physiological significance of alterations in volley duration. The dynamics of LH pulses in ovariectomized animals were observed in a number of experimental circumstances in which MUA volley duration was reduced from a maximum of 23 min to a minimum of 4 min without significantly altering their frequency. The magnitude of each LH pulse was assessed by calculating the area under the curve delineated by the time course of LH above baseline. In eight experiments, a linear regression of these values on volley duration failed to reveal a significant correlation between MUA volley duration and the magnitude of LH pulses. These results suggest that all of the GnRH secreted per pulse is released at the onset of each MUA volley, the remainder of the increase in electrical activity having no further action on GnRH secretion, although effects on other systems cannot be excluded.

Activity Cycles↗

Corticotropin-releasing factor and gonadotropin-releasing hormone pulse generator activity in the rhesus monkey. Electrophysiological studies.

The effect of corticotropin-releasing factor (CRF) on the hypothalamic gonadotropin-releasing hormone (GnRH) pulse generator, the central neuronal system governing pulsatile pituitary luteinizing hormone (LH) secretion, was studied electro-physiologically in 6 ovariectomized rhesus monkeys bearing bilateral arrays of recording electrodes implanted in the mediobasal hypothalamus. 'Volleys' of increased multiunit activity (MUA) were recorded for 6-10 h in animals placed in primate chairs. The circulating concentrations of LH and cortisol were determined by radioimmunoassay in blood samples taken every 10 min for 3-4 h prior to the administration of CRF (200 micrograms, i.v.) and for 3-6 h thereafter. CRF resulted in a significant decrease in the frequency of pulse generator activity in 4 of 6 animals, a significant decrease in the duration of MUA volleys and a rise in circulating cortisol levels in all 6 monkeys. Treatment with metyrapone (30 mg/kg, i.m.), an inhibitor of adrenal steroidogenesis that prevented the CRF-induced rise in serum cortisol levels, did not reverse the inhibitory effects of CRF on the frequency or duration of MUA volleys. The opiate antagonist naloxone (0.8 mg/kg, i.v., 10 min prior to CRF followed by 0.8 mg/kg/h infusion) blocked the effects of CRF on MUA volley frequency in 2 of 3 animals, but failed to block the effect of CRF on MUA volley duration, suggesting that endogenous opioids may mediate the action of CRF on pulse generator frequency but not on duration.

Animals↗

Duration and frequency of multiunit electrical activity associated with the hypothalamic gonadotropin releasing hormone pulse generator in the rhesus monkey: differential effects of morphine.

The effects of morphine on the frequency and duration of the characteristic bursts or 'volleys' of multiunit electrical activity (MUA) associated with pulsatile pituitary luteinizing hormone (LH) secretion were studied in unanesthetized ovariectomized rhesus monkeys bearing bilateral arrays of electrodes implanted in the mediobasal hypothalamus. Morphine administration resulted in a dose-dependent decrease in MUA volley duration and frequency. When morphine was infused at 10 micrograms/kg/h, the inhibiting effect on volley duration was observed without a change in volley frequency. It is concluded that the frequency and duration of hypothalamic MUA volleys associated with pulsatile LH secretion may be independently regulated.

Animals↗

The electrophysiology of the GnRH pulse generator in the rhesus monkey.

The electrophysiological correlates of hypothalamic GnRH pulse generator activity in the rhesus monkey are described. They are characterized by abrupt increases in neuronal multiunit electrical activity (volleys) that are coincident with the initiation of LH pulses in the peripheral circulation. Both the frequency and duration of these volleys can be varied by a variety of endocrinological, pharmacological and environmental perturbations. Their activity throughout the rhesus monkey menstrual cycle as assessed by radiotelemetry is described.

Animals↗

The neuroendocrine control of ovulation.

In all species studied to date, follicular development is the consequence of pulsatile gonadotrophin stimulation occasioned by a GnRH pulse generator located in the mediobasal hypothalamus. This pulse generator operates within a narrow frequency range established by a neuronal system which, in the higher primates including man, discharges with a period of approximately 1 h. This frequency does not appear to change significantly as the follicular phase of the menstrual cycle progresses and plasma oestradiol concentrations rise. When these exceed a defined threshold for approximately 36 h, the pre-ovulating gonadotrophin surge is initiated and ovulation ensues some 24 h later, the so-called positive feedback action of oestrogen. This sequence of events can be replicated experimentally in subjects with various hypothalamic lesions which abolish endogenous GnRH production, simply by the administration of exogenous GnRH at an unchanging physiological frequency. This implies that the site of the positive feedback action of oestradiol is the pituitary gland and that GnRH plays but a permissive role in the initiation of the mid-cycle gonadotrophin surge. This process can be disrupted by a number of inputs which perturb the proper functioning of the hypothalamic GnRH pulse generator.

Animals↗

Unexpected responses of the hypothalamic gonadotropin-releasing hormone "pulse generator" to physiological estradiol inputs in the absence of the ovary.

In the rhesus monkey (Macaca mulatta), the frequency of pulsatile gonadotropic hormone release is relatively constant in the face of widely varying levels of estradiol (E2) in the peripheral circulation--e.g., in the course of the follicular phase of the menstrual cycle and after ovariectomy. This suggests that modulation of the hypothalamic gonadotropin-releasing hormone (GnRH) "pulse generator" by this steroid is not of major physiological importance. Herein is described the unexpected inhibition or total blockade of the electrical activity of this pulse generator in ovariectomized monkeys by physiological levels of exogenous E2. This inhibition began 2-4.5 hr after the initiation of E2 infusion and was noted 1 to 3 weeks after subcutaneous implantation of E2-containing capsules. Pulse generator activity was also arrested during the initiation and subsequent development of estrogen-induced surges of luteinizing hormone. We propose that this inhibition of hypothalamic GnRH pulse generator activity by E2 in ovariectomized monkeys reflects the absence of an ovarian factor that normally protects this neuronal system from the inhibitory action of estrogen during the menstrual cycle.

Action Potentials↗

On the short-loop feedback regulation of the hypothalamic luteinizing hormone releasing hormone 'pulse generator' in the rhesus monkey.

The characteristic electrophysiological manifestations of LHRH 'pulse generator' activity were unaffected by prolonged elevations in plasma LH concentrations achieved by the administration of a long-acting LHRH agonist. These findings do not lend support to the existence of a 'short-loop feedback' inhibition of LHRH 'pulse generator' activity by LH.

Animals↗

The effect of morphine on the electrophysiological activity of the hypothalamic luteinizing hormone-releasing hormone pulse generator in the rhesus monkey.

In ovariectomized rhesus monkeys, the electrophysiological manifestation of luteinizing hormone-releasing hormone (LHRH) pulse generator activity was arrested by morphine and reinitiated by naloxone. These responses were noted within 1 and 2 min, respectively, after the intravenous injection of the drugs. Naloxone given alone had no effect. These results support the view that opioids modulate pulsatile gonadotropin release by an action on the hypothalamic LHRH pulse generator.

Animals↗

Hypothalamic localization of multiunit electrical activity associated with pulsatile LH release in the rhesus monkey.

Abrupt increases in electrical activity can be recorded from the medial basal hypothalamus of the rhesus monkey and these bursts are correlated with a pulse of LH release from the anterior pituitary. In this study we have localized the tips of the electrodes from which such electrical activity can be recorded and have attempted to correlate the placement with the presence of immunoreactive LHRH neurons and axons. Electrodes associated with bursting activity were found as far rostral as the suprachiasmatic nucleus and as far caudal as the premammillary nuclei. The majority were found in the medial basal hypothalamus including the region of the arcuate nucleus, the retrochiasmatic zone and the dorsal aspect of the median eminence. The tips of the electrodes were associated with either LHRH neurons or axon bundles. Negative electrodes (those not associated with bursting electrical activity) which did not exit through the ventral surface of the brain nor enter the ventricle were found in the same regions of the hypothalamus as positive electrodes. Although it is tempting to associate the bursting electrical activity with the LHRH neurosecretory system, such a correlation cannot yet be made.

Animals↗

Electrophysiological manifestation of luteinizing hormone-releasing hormone pulse generator activity in the rhesus monkey: influence of alpha-adrenergic and dopaminergic blocking agents.

Characteristic increases in neuronal electrical activity associated with the initiation of each LH pulse were recorded from ovariectomized rhesus monkeys bearing multiple chronic electrodes in the medial basal hypothalamus. These electrophysiological manifestations of hypothalamic LHRH pulse generator activity were inhibited by the alpha-adrenergic blocker phentolamine or the alpha 1-adrenoceptor blockers phenoxybenzamine and prazosin. At the dosages used, the effects of single injections of these drugs ranged from a reduction in the frequency of LHRH pulse generator activity to its complete arrest. This was faithfully reflected in the pattern of pulsatile LH discharges. The dopaminergic blocking agent metaclopramide similarly reduced the frequency of the pulse generator or arrested its activity altogether. The alpha 2-adrenoceptor blocker yohimbine had no demonstrable effect on hypothalamic electrical activity at the doses studied. These findings support the view of a central action of alpha 1-adrenergic and dopaminergic blockade on LHRH pulse generator activity and the concept that central adrenergic and dopaminergic inputs can modulate the frequency of the LHRH pulse generator.

Adrenergic alpha-Antagonists↗

The role of prolactin in the regulation of the primate corpus luteum.

The role of PRL in the control of corpus luteum (CL) function was examined in hyperprolactinemic and euprolactinemic female rhesus monkeys with hypothalamic lesions in which ovulatory menstrual cycles were induced by the pulsatile administration of GnRH, in intact postpartum monkeys nursing their infants, as well as in animals treated with bromocriptine. In the lesioned monkeys, neither markedly elevated nor undetectable (bromocriptine-suppressed) PRL levels influenced the time courses of postovulatory plasma progesterone concentrations compared to those in monkeys with normal PRL concentrations. Plasma progesterone concentrations in the hyperprolactinemic monkeys, however, did not decline to undetectable levels at the end of the luteal phase, as is the case in normal animals, but remained slightly elevated for extended periods, a pattern characteristic of lactating animals. In both lesioned and postpartum animals, bromocriptine administration or removal of suckling infants from the latter resulted in complete functional luteolysis. These observations suggest that PRL does not play a significant role in the control of the CL of the menstrual cycle, but that in high concentrations, this hormone can partially maintain previously established CL.

Animals↗