Caloric Intake Stimulates Growth Hormone Secretion in Food-Deprived Rats with Anterolateral Deafferentation of the Medial Basal Hypothalamus or Administered Antiserum to Somatostatin

In rats, food deprivation inhibits episodic growth hormone (GH) secretion. On the basis of previous studies, we hypothesized that during a recovery from prolonged fasting, caloric intake stimulates the release of GH-releasing factor (GRF) and this process does not depend on the specific macronutrients in the meal, while protein in the meal acts to restore characteristic ultradian rhythmicity of GH secretion. To test this hypothesis, the effect of caloric intake on GH secretion was examined in fasted adult male Wistar rats devoid of somatostatin (SS) influence on GH secretion either by anterolateral deafferentation (ALC) of the medial basal hypothalamus (MBH) or administration of anti–SS goat serum (ASS). Rats were provided with an indwelling right atrial cannula and were deprived of food for 72 h. ALC was performed 2 weeks prior to the study. ASS was given i.v. 8 h and 7 h prior to refeeding, respectively. Serial blood specimens were collected every 10 min. fn rats with ALC (ALC rats) or rats given ASS (ASS rats), the blood GH level revealed irregularly occurring small fluctuations, instead of the usual high bursts and low trough level. The baseline GH level and the mean GH level of fasted ALC rats or fasted ASS rats were significantly lower than those of fed ALC rats or fed ASS rats. Feeding the isocaloric mixed meal, the protein meal or the protein-deficient meal increased the GH pulse frequency, the pulse amplitude, the baseline GH level and the mean GH level in 72–h fasted ALC rats. These changes in GH secretory pattern persisted during the period of observation and were independent of the type of meal ingested. Following feeding the mixed meal, similar changes in the GH secretory pattern demonstrated in 72-h fasted ALC rats were also observed in 72–h fasted ASS rats, suggesting that the stimulation of GH secretion following caloric intake is not limited to ALC rats. Since the influence of SS on GH secretion has been largely eliminated in ALC or ASS rats, it is highly unlikely that the augmentation of GH secretion following feeding after prolonged food deprivation was the consequence of inhibition of SS secretion. Although GRF measurement was not performed, it is conceivable that the signal of caloric intake is conveyed to the MBH and acts to stimulate GRF release.     

Effects of Somatostatin on the Growth Hormone-Releasing Factor-Induced Growth Hormone Secretion in Rats with Electrical and Chemical Inhibitions of Endogenous Growth Hormone-Releasing Factor and Somatostatin

The episodic pattern of growth hormone (GH) secretion of the male rat was simulated in rats exhibiting impaired GH-releasing factor (GRF) and Somatostatin (SRIF) secretion, by administering various combinations of human GRF-(1–44)NH₂ (hGRF) and SRIF. Electrical lesions of the arcuate nucleus resulted in a marked decrease in the amplitude of GH secretory bursts, while the administration of cysteamine (200 mg/kg) did not change the GH secretory profile in rats with arcuate nucleus lesions. Immunohistochemical examinations revealed a marked decrease of GRF and SRIF immunoreactivity in the median eminence of the cysteamine-treated rats with arcuate nucleus lesions. The intravenous injection of 5 μg of hGRF every 3 h caused equivalent surges of GH in the cysteamine-treated rats with arcuate nucleus lesions. The additional infusion of 4 μg/h of SRIF during the trough periods of GH secretion did not affect the amplitude of the GH surges. Hourly injection of 5 μg of hGRF caused transient desensitization to hGRF. Interestingly, the additional infusion of 4 μg/h of SRIF every 3 h enhanced the amplitude of the GH bursts induced by the fourth and the seventh hGRF injections. However, the more frequent injection of 5 μg of hGRF every 30 min caused constant desensitization to hGRF with time, and the additional infusion of 4 μg/h of SRIF every 3 h did not change the attenuated responses to hGRF. These results indicated that the simultaneous administration of hourly GRF and continuous SRIF with brief pauses was most effective for producing high GH peaks. This simulation model suggests that SRIF may play an important role not only in the production of GH troughs, but also in the maintenance of GH surges with distinct peaks in the male rat.     

Different Effects of Somatostatin on in vitro Growth Hormone Release in Two Porcine Breeds with Different Growth Rates1

A perifusion system of anterior pituitary (AP) tissue was used to investigate the temporal interaction of growth hormone-releasing factor (GRF) and somatostatin (SRIF) in the control of GH secretion in two pig breeds, Göttingen Miniature Pig (GMP), a small obese breed, and German Landrace (GLR), a conventional lean breed. AP tissue pieces derived from sexually mature ovariectomized animals were perifused (6 replicates per treatment) and fractions were collected at 10 min intervals. Basal GH release (ng-mP^-1 mg^-1 AP) in GLR was twice that of GMP (P < 0.001). Exposure to 10 min pulses of 1 nM GRF repeated 3 times at 2 h intervals resulted in rapid stimulatory GH responses (area under the curve) which became attenuated (P < 0.05) over time in GMP but not in GLR. Surprisingly, during and following the exposure of AP tissue from GMP to 10-, 20-, or 40-min pulses of 10 nM SRIF alone, GH release was markedly stimulated (P < 0.05), while AP tissue from GLR only showed a weak rebound GH release after SRIF pulses. With AP tissue from GLR low concentrations (0.1 nM SRIF) amplified GRF-induced GH release, whereas 1 nM or 10 nM SRIF inhibited GRF-induced GH release. However, concomitant exposure of AP tissue from GMP to 0.1, 1 or 10 nM SRIF during a GRF pulse markedly enhanced the GH response (P < 0.05), compared to 1 nM GRF alone, except for 1 nM SRIF which inhibited the GH response to the first GRF pulse. Thus the presence of SRIF, and not only its withdrawal, is an important factor in setting the timing and duration of GH pulses in both breeds. In GLR the concentration of SRIF is more important than the duration and/or type of SRIF pulse. In contrast, in GMP type and/or duration of SRIF pulses seem to be crucial to optimize pulsatile GH release and even determine peak height of GH pulses caused by GRF. These findings indicate clear breed differences in the role of SRIF and in the control of GH release by the interplay of GRF and SRIF. The ‘paradoxical’ effect of SRIF suggests that the role of SRIF is much more complex than that of a mere inhibitor and whose real role could be a modulator either of GH pulse and/or GRF action on GH release.     

Ovarian Steroid Regulation of Pulsatile Luteinizing Hormone Release During Early Gestation in the Rat

The object of this study was to examine ovarian regulation of pulsatile luteinizing hormone (LH) secretion during early gestation. This was done primarily by analyzing pulsatile LH release in rats that were either sham ovariectomized (OVX) on Day 7 of pregnancy, implanted with empty Silastic capsules, and bled on Day 8, or OVX on Day 7, immediately implanted with Silastic capsules producing plasma levels of estradiol and/or progesterone characteristic of Day 7 to 8 of pregnancy, and bled on Day 8. In addition, the role of progesterone in regulating pulsatile LH secretion was also examined by administration of the progesterone receptor antagonist, RU486, on Day 7 and examining pulsatile LH release on Day 8 of pregnancy. OVX caused a marked increase in LH pulse amplitude and frequency within 24 h. Replacement with physiological plasma levels of estradiol or progesterone alone had no suppressive effect on this OVX-induced increase in pulsatile LH secretion. Restoration of physiological plasma levels of both estradiol and progesterone returned LH pulse amplitude to values seen in sham OVX controls, and prevented the OVX-induced increase in LH pulse frequency. The group mean LH pulse frequency tended to be less in estradiol + progesterone-treated rats than in sham OVX controls, but this difference was not statistically significant. RU486 blocked uterine progesterone receptors as evidenced by endometrial hemorrhaging. In agreement with the OVX + steroid replacement data, RU486 administration also resulted in increases in LH pulse amplitude and frequency. These data demonstrate that the frequency and amplitude of LH pulses on Day 8 of gestation are held in check by negative feedback signals coming from the ovary. Neither steroid alone exerts any suppressive influence over pulsatile LH secretion during early gestation, but both steroids acting together exert a prominent negative feedback regulation on the pulsatile LH release process.     

Ultradian Rhythm of Growth Hormone Secretion in Unrestrained,Conscious Male Rabbits: Role of Endogenous Somatostatint†

The profiles of growth hormone (GH) secretion were examined by obtaining serial blood samples every 15 min for a 5 to 24 h observation period from freely-moving, conscious male rabbits chronically implanted with a right atrial cannula. The effects of restraint or surgical stress on GH secretion were also investigated in these animals. Four days after cannulation of the right atrium, plasma GH levels remained low without oscillation, during a 5 h observation period (1100 to 1600 h) with the mean (± SEM) value of 1.6±0.2 ng/ml. Individual rabbits exhibited a spontaneous, pulsatile GH secretion 7 days after the surgery. Mean 6 h GH levels were 5.6 ± 0.8 ng/ml at 7 days after the surgery, 6.3 ± 0.6 ng/ml at 14 days and 7.0 ± 1.2 ng/ml at 28 days. Therefore, the animals, 7 to 14 days after cannulation, were used to analyse the pulsatile pattern of GH secretion throughout 6 to 24 h. Two episodes of 45 min immobilization stress, separated by 75 min, caused a complete suppression of the spontaneous GH secretion (mean 6 h GH levels, 2.2 ± 0.1 ng/ml vs control, 5.0 ± 0.5 ng/ml, P<0.01). No surges appeared after the first restraint stress. In 14 non-treated rabbits, plasma GH levels fluctuated in an episodic manner throughout the study with the peaks of 14.2 + 0.7 ng/ml, the nadirs of 2.6 ± 0.2 ng/ml and the peak to peak intervals of 2.20 ± 0.17 h. The iv administration of normal goat λ-globulin (NGG) affected neither GH secretory patterns nor baseline levels of plasma GH. In contrast, the iv administration of anti-sornatostatin goat λ-globulin (ASG) caused a significant increase in the amplitude of plasma GH peaks (38.8±1.9 vs NGG-treated, 13.7 ± 0.8 ng/ml, P<0.001) as well as the trough level (13.5 ± 0.6 vs NGG, 2.9 ± 0.1 ng/ml, P<0.001) during a 24 h observation period. Also, ASG treatment increased numbers of plasma GH peaks per day (18.8±2.7 vs NGG, 12.2 ± 0.8, P < 0.05) with concomitant shortening of the peak to peak interval (1.25 ± 0.10 vs NGG, 2.03±0.12h, P<0.01). These findings suggest: 1) that GH is episodically secreted throughout the day in conscious male rabbits, 2) that surgical and restraint stresses suppress the spontaneous GH secretion, and 3) that endogenous somatostatin might rather play a tonic inhibitory role in GH release in conscious male rabbits, since ASG treatment resulted in sustained marked increases of plasma GH levels irrespective of the stage in GH pulsatile rhythm.     

Adrenergic and Noradrenergic Regulation of Pulsatile Luteinizing Hormone Release*

The object of this study was to further define the roles of both norepinephrine (NE) and epinephrine (EPIN) in regulating pulsatile luteinizing hormone (LH) release in 4-day ovariectomized rats, in particular to examine the effect of decreasing NE synthesis on pulsatile LH secretion in animals with already greatly depleted levels of brain EPIN. Rats were injected ip with vehicle or drug at -27, -20, -5 and - 3 h relative to the onset of a 3-h blood sampling period. Hypothalamic-preoptic area (HPOA) levels of NE and EPIN were determined by high-performance liquid chromatography. Compared to controls, FLA-63 (25 mg/kg, a dopamine-ß- hydroxylase inhibitor), given at - 3 h, produced 50% and 22% declines in HPOA-NE and EPIN, respectively, and reductions in pulse amplitude and frequency. LY134046 (50 mg/kg, a phenylethanolamine N-methyltransferase inhibitor), given at - 27, - 20 and - 5 h, or -27, -20, -5 and -3 h, produced no change in NE, 88% and 86% declines in EPIN, respectively, and reductions in pulse frequency only. Each LY134046 treatment protocol produced the same decline in EPIN and pulse frequency. Thus, EPIN levels were maximally decreased by three LY134046 injections. When rats were given LY134046 at -27, -20 and -5 h, and FLA-63 at -3 h, compared to rats treated with LY134046 alone, there was no further decrease in HPOA-EPIN (82% decline), a 46% decline in NE, a further reduction in pulse frequency and a reduction in pulse amplitude. This further suppression of LH release must be due to a reduction in HPOA-NE levels since no further decrease in EPIN levels occurred. These data demonstrate within the same animal that NE and EPIN are both stimulatory to pulsatile LH release. NE stimulates the amplitude and frequency, and EPIN stimulates the frequency of pulsatile LH secretion.     

Locus ceruleus lesions block pulsatile LH release in ovariectomized rats

Luteinizing hormone (LH) secretion during the reproductive cycle and in ovariectomized (OVX) rats is pulsatile and this pattern of secretion is determined by intermittent discharges of LH-releasing hormone (LHRH) into the hypophysial portal vessels. LHRH secretion is probably controlled by prior pulsatile norepinephrine (NE) release. The locus ceruleus (LC) is an important source of NE to the LHRH neurons. We have shown previously that LC lesions block the preovulatory LH surge and ovulation and also cause a decrease in plasma LH concentrations in OVX rats. The possible role of the LC in regulating pulsatile LH release has not been explored. Therefore, the aim of this work was to investigate, in OVX rats, the effects of LC lesions on pulsatile LH secretion. LC lesions were produced in adult female rats three weeks after OVX. On the next morning, the jugular vein was catheterized and, on the afternoon of the same day, blood samples (0.3 ml) were withdrawn every 5 min, during 90 min, from conscious freely moving rats. Plasma LH was measured by radioimmunoassay. LC lesions greatly suppressed pulsatile LH secretion by decreasing both LH pulse frequency and amplitude. The basal as well as total secretion of LH were also decreased. This inhibitory effect of the lesions was observed only when at least 50% of the nucleus was destroyed. Data from sham-operated animals as well as those with less than 50% destruction of the LC did not differ from those of the control rats without brain lesions. Since LC lesions induce a decrease in NE content in the preoptic area and median eminence, the inhibition of pulsatile LH release in ovariectomized rats with LC lesions occurs presumably as result of decreased pulsatile NE release into these areas of the brain that decreases both the frequency and the amount of LHRH released per pulse.     

Monosodium glutamate: Acute and chronic effects on rhythmic growth hormone and prolactin secretion, and somatostatin in the undisturbed male rat

The present investigation was designed to determine the chronic effects of neonatal monosodium glutamate (MSG) administration (4 g/kg s.c.) and the acute effects of MSG (1 g/kg i.p.) on episodic growth hormone (GH) and prolactin (PRL) secretion and brain somatostatin (SRIF) in unanesthetized, chronically cannulated male rats.Adult rats showed the typical physical characteristics that result from neonatal MSG administration. Analysis of episodic GH secretion showed a significant reduction in: (1) the amplitude of GH secretory peaks; and (2) the mean 5.5-h plasma level of GH. Bursts of plasma PRL were inhibited by MSG, but the mean 5.5-h plasma levels were not affected. SRIF concentrations in the medial basal hypothalamus were reduced by 60% after neonatal MSG. Acute administration of MSG to adult rats caused an immediate, long-lasting suppression of rhythmic GH secretion and a rapid, transient release of PRL.These results suggest: (1) neonatally administered MSG causes a marked disturbance in episodic GH and PRL secretion in adult rats; (2) MSG induces a decrease in hypothalamic SRIF and possibly GH-releasing factor; and (3) the acute effects of MSG on GH and PRL may be due to the inhibition and/or excitation of a complex neuronal network involving monoaminergic and peptidergic systems.     

Analysis of estradiol-independent and -dependent endogenous opioid peptide suppression of pulsatile LH release between the mornings of diestrus 2 and proestrus in the rat estrous cycle

The aim of this study was to analyze possible estradiol (E2)-independent and -dependent endogenous opioid peptide (EOP) suppression of pulsatile LH release between the mornings of diestrus 2 (D2) and proestrus by examining the LH response to naloxone infusions in the presence or absence of proestrous levels of E2. Pulsatile LH secretion remained unchanged between D2 and proestrus but mean blood LH levels, pulse amplitude and frequency increased within 24 hr following ovariectomy on D2. This increase was due in large part to the loss of E2 negative feedback, since restoration of physiological proestrous E2 levels returned LH pulse frequency to proestrous a.m. levels and greatly reduced pulse amplitude. In ovariectomized rats lacking E2 negative feedback, continuous infusion of the EOP receptor antagonist naloxone (0.5 and 2 mg/kg/hr) caused a further increase in pulse amplitude and frequency. This naloxone-induced increment in pulsatile LH release was exerted via centrally located EOP receptors since naloxone did not alter pituitary responsiveness to LHRH, and its stimulatory action on pulsatile release was diminished by simultaneous infusion with morphine. Naloxone also increased pulsatile LH release in E2-treated animals. The naloxone-induced increments in LH pulse amplitude were the same in the presence or absence of E2 negative feedback. Moreover, the increments in amplitude produced by naloxone in E2-treated rats were significantly less than those resulting from the combination of ovariectomy plus naloxone infusion in empty capsule-implanted rats. These data indicated that naloxone infusion in E2-implanted animals blocked an E2-independent EOP suppression of this parameter of pulsatile release.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pulsatile Luteinizing Hormone Secretion in the Rat: Methodological Aspects of Cannulation Route and Sampling Intensity

We have investigated some requirements for valid estimates of pulsatile luteinizing hormone (LH) secretion in the castrate male rat. The choice of cannulation route and the minimal sampling intensity required to produce stable, unbiased estimates of parameters of pulsatile LH secretion were examined. Castrate mature male Wistar rats with unilateral ligation of a carotid artery (an inevitable consequence of carotid artery cannulation) had a 47% reduction in mean LH levels (1.8 vs 3.4 ng/ml, P< 0.011) attributable to a 40% reduction in LH pulse frequency (6.2 vs 10.4 peaks/6 h P<0.006) without change in LH pulse amplitude or other parameters of pulsatile LH secretion. In another study, a range of sampling intensities were examined to derive estimates of pulsatile LH secretion. Parameter estimates derived from the most intensive regime (q10 min sampling for 6 h) were compared with estimates of pulsatile LH secretion parameters determined from less intensive derivative sub-series composed of either fixed sampling frequency, but variable total study duration (i.e. the first 1, 2, 3, 4 and 5 h at 10 min intervals), or a fixed total study duration but variable sampling frequency. LH pulse frequency (or its inverse, interpulse interval) was quite sensitive to sampling intensity whereas pulse amplitude, maximum, minimum and overall mean LH were estimated accurately at even relatively low sampling intensity (q20 min for one h duration). Lower frequency sampling (q20 min) gave marked underestimates of LH pulse frequency (4.9 vs 10.4 peaks/6 h, P<0.05) compared with q10min sampling over a 6h study. These results indicated that for stable and unbiased estimates of pulsatile LH secretion in the castrate mature male rat it is necessary (but may not be sufficient) to (1) avoid carotid artery cannulation and (2) undertake blood sampling at intervals of not more than 10 min for at least 3 h.     

Rostromedial septal area controls pulsatile growth hormone release in the golden hamster

Limbic forebrain inhibits growth and growth hormone (GH) secretion in mature golden hamsters as shown by acceleration of growth and increases in serum GH concentrations following the electrolytic lesions of septum, transection of the hippocampus and surgical separation of these two regions. The growth-inhibitory function of this circuit is most probably mediated by somatostatinergic (SRIF) neurons. Such lesions induce hypoactivity possibly due to damage to endogenous opiatergic (EOP) neurons. EOP neurons facilitate spontaneous running in hamsters and mediate exercise-induced acceleration of growth and GH pulses. The coincidence of hypoactivity and growth acceleration after such lesions suggested the coexistence of SRIF and EOP fibers within the growth-inhibitory limbic forebrain circuit which control the rate of growth in mature hamsters by the variable inhibition of SRIF neurons by the EOP neurons. This hypothesis posits that accelerated growth is due to increased GH pulse frequency, and hypoactivity due to damage to EOP neurons, and was tested in this study by measuring pulsatile GH release (and as a measure of specificity, pulsatile prolactin release) in the presence and in the absence of opiate-receptor blocker naloxone in 21 female hamsters which sustained electrocoagulative lesions of rostromedial septum and 30 hamsters subjected to control surgery. Lesions doubled GH but not PRL pulse frequency, neither of which was affected by naloxone. Results support the hypothesis that opiatergic neurons facilitate pulsatile GH release by inhibiting the action of somatostatin neurons.     

Effects of castration on LH-RH patterns in intrahypophysial microdialysates

A microdialysis system was used to monitor LH-RH patterns in the extracellular fluid of the adenohypophysis of testes-intact and short-term castrate rats. Male rats received guide cannulae implants fitted with stylets that extended into the anterior pole of the anterior pituitary gland. At the same time, animals were either castrated or received sham surgeries. On day 4 following surgeries, microdialysis probes were inserted into the guide cannulae and artificial CSF was pumped through the system at a flow rate of 2.5 μl/min. Continuous samples were obtained from each animal over 5- or 10-min intervals throughout 4–7 sessions. Placements of probe tips were verified by histological examination of stained tissue sections. In vitro tests of microdialysis probe performance revealed an exchange rate of 4% at the 2.5 μl/min flow rate. In vivo patterns of LH-RH in microdialysates obtained from sham-operated and castrate rats were pulsatile, as determined by the computer algorithm ULTRA. Pulses of LH-RH occured at a higher frequency (P < 0.05) in the castrates (1.30 ± 0.26pulses/h n = 6) versus the sham-castrates (0.87 ± 0.06pulses/h, n = 11). Mean LH-RH pulse amplitude (castratesδ0.24 ± 0.03 pg,testes-intactΔ 0.42 ± 0.06pg) and mean LH-RH levels (castrate0.37 ± 0.04 pg/10 min, intact0.48 ± 0.06 pg/10 min), however, were not significantly changed by castration (Δ =difference between trough and peak LH-RH value of an LH-RH pulse). Our observations are in direct agreement with previous LH-RH release measurements using hypothalamic push-pull perfusion, and support the contention that negative feedback actions of gonadal hormones are mediated, in part, via suppression of LH-RH pulse generation. These findings also confirm the usefulness of microdialysis in monitoring hypothalamic neurosecretory signals following delivery via the portal vasculature, viz. in the extracellular spaces of the anterior pituitary gland.     

Effects of lesions in the periventricular nucleus of the preoptic-anterior hypothalamus on growth hormone and thyrotropin secretion and brain somatostatin

Two experiments were performed to study the role of somatostatin (SRIF) neurons of the preoptic-anterior hypothalamic area (PO-AHA) in regulating growth hormone (GH) and thyrotropin (TSH) secretion in rats. Small lesions were placed in the periventricular (PV) zone and blood was collected at 24 h and 15 days after surgery. Blood samples were obtained at 3 min and at 15 min after ether exposure for assessing non-stress levels, respectively, of plasma GH and TSH. Non-stress blood samples were also collected at decapitation at 4 weeks. The brains from the first experiment were dissected and processed for measuring SRIF content in several regions. At 24 h and 15 days, non-stress GH and TSH levels were significantly elevated in rats with PV lesions. Stress-induced decrements in GH levels persisted in all groups. Although non-stress plasma GH and TSH levels returned to normal in lesioned rats at 4 weeks, SRIF content was decreased 83% in the median eminence and 33% in the hypothalamus. These results show that discrete lesions in the PV zone of the PO-AHA cause transient elevations in non-stress secretion of GH and TSH and that normal levels of such secretion can be reinstated despite reductions of SRIF in the median eminence and hypothalamus.     

Effect of dopamine receptor blockade or norepinephrine synthesis inhibition on acute, ovariectomy-induced increases in pulsatile luteinizing hormone release in the rat

The initial aim of the present studies was to examine the influence of blockade of dopamine (DA) receptors with pimozide or inhibition of norepinephrine (NE) synthesis with U-14,624 on acute, ovariectomy (OVX)-induced changes in pulsatile LH release. Either treatment instituted at the time of OVX suppressed or inhibited the rapid increase in LH pulse amplitude and frequency normally occurring within 24 hr following ovarian removal on diestrus 1. While administration of pimozide at either 24 hr or 48 hr following OVX suppressed pulsatile LH release by selectively reducing LH pulse frequency, by 8 days following OVX pimozide failed to exert any effect on LH pulse frequency and therefore on pulsatile LH secretion. To determine if there was a transient critical period following OVX of at least 2 days but less than 8 when endogenous DA was excitatory to pulsatile LH release, piribedil (a DA receptor agonist) was given 24 hr following OVX. Rather than increase LH secretion, piribedil markedly suppressed pulsatile LH release indicating that DA does not stimulate LH secretion in acutely ovariectomized rats. These experiments indicate that

1. (1) NE is involved in stimulating the acute, OVX-induced increase that occurs in pulsatile LH release;

2. (2) DA receptorblockade by pimozide has a differential effect on pulsatile LH secretion which depends on the time following OVX when the compound is administered;

3. (3) this differential effect cannot be explained by a transient critical period of a few days duration following OVX during which DA is excitatory to pulsatile LH release.

Intracerebroventricular administration of the rat growth hormone (GH) receptor antagonist G118R stimulates GH secretion: evidence for the existence of short loop negative feedback of GH

Pulsatile growth hormone (GH) secretion is regulated by three hypothalamic factors, growth hormone-releasing hormone (GHRH), somatostatin and the natural ligand for the GH secretagogue receptor (Ghrelin). These factors and their effects are, in turn, affected by short loop feedback of GH itself. To test the hypothesis that hypothalamic GH receptors are involved in the ultradian rhythmicity of pituitary GH secretion, the rat GH receptor antagonist (G118R) was administered to adult male rats by intracerebroventricular (i.c. v.) injection and the effects on spontaneous GH secretion were studied. Normal saline was administered i.c.v. to eight control rats. Mean GH concentrations increased significantly in the rat treated with G118R compared to rats that received normal saline. The pulse amplitude rose by a mean of 33.3 ng/ml and the total area under the curve increased by a mean of 15 061 ng/ml x min. The number of GH peaks did not change significantly following G118R. These data suggest that GH regulates its own secretion by acting directly on hypothalamic GH receptors.     

A Rapid Suppressive Effect of Estrogen in the Paraventricular Nucleus on Pulsatile LH Release in Fasting-Ovariectomized Rats

The paraventricular nucleus (PVN) and A2 are novel estrogen feedback sites where estrogen may modulate the neural response to adrenergic inputs during fasting. In the present study, the effects of local estradiol (E₂) perfusion through a microdialysis probe placed in the PVN or A2 on pulsatile luteinizing hormone (LH) secretion and on norepinephrine (NE) release in the PVN were examined in 48-h fasting ovariectomized (OVX) rats to determine whether local estrogen administered in the PVN or A2 rapidly inhibits LH secretion during fasting and whether this inhibition is mediated by an increase of NE release in the PVN. Five days after ovariectomy, animals (n=5 per group) stereotaxically implanted with a guide cannula for microdialysis in the PVN (experiment 1) or both PVN and A2 (experiment 2) were deprived of food for 48  h. Blood samples and dialysates were then collected every 6  min for 3  h and every 12  min (experiment 1) or 20  min (experiment 2) for 3  h, respectively. The PVN or A2 was perfused with E₂ (5  ng/ml in artificial cerebrospinal fluid) through a microdialysis probe after the first hour of sampling. E₂ perfusion in the PVN caused a rapid and significant suppression of mean plasma LH levels and LH pulse frequency in fasting rats but no changes in unfasting animals. NE release in the PVN was not affected by the local E₂ perfusion of the PVN in either fasting or unfasting groups. This perfusion in A2, however, did not cause any apparent changes in plasma LH and perfusate NE levels in the PVN and A2. The present results indicate that estrogen feedback action at the PVN suppresses LH secretion rapidly during fasting and does not involve an increase of NE release in the PVN.     

Effects of Various Types of Hypothalamic Deafferentation on Luteinizing Hormone Pulses in Ovariectomized Rats

The pulsatile luteinizing hormone (LH) secretion in chronically ovariectomized rats bearing various types of hypothalamic deafferentation was examined. Ovariectomized rats were subjected to complete, anterolateral or anterior hypothalamic deafferentation and bled every 6 min for 3 h through an indwelling atrial cannula 5 days after the brain surgery. Another group of ovariectomized animals was subjected to posterior-anterior hypothalamic deafferentation (PAD), which cut off the anterior part of the arcuate nucleus from the mediobasal hypothalamus, and bled 1, 3 or 5 weeks after the deafferentation. Coronal sections of the brain were immunostained with anti-LH-releasing hormone (LHRH) serum. The pulsatile LH secretion was observed in rats bearing anterior, anterolateral or complete hypothalamic deafferentation and these types of deafferentation did not affect the frequency of LH pulses. The mean LH level during the 3-h sampling period and the amplitude of LH pulses decreased as the incision extended postero-laterally. Rats bearing PAD showed an irregular fluctuating pattern in plasma LH concentration 1 week after PAD. Parameters of LH pulses were restored with time after PAD. This suggests that the system generating LHRH pulses severed by PAD had been reorganized. LHRH-immunopositive neuronal fibres were found in the external layer of the median eminence in the rats bearing any type of deafferentation. The present results suggest that the frequency of LH pulses could be controlled by the LHRH pulse generator, which consists of non-LHRH neurons and is located in the mediobasal hypothalamus.     

Intracerebroventricular administration of melanin-concentrating hormone suppresses pulsatile luteinizing hormone release in the female rat

Melanin-concentrating hormone (MCH) has been reported to be involved in the regulation of feeding behaviour in rats and mice. Because many neuropeptides that influence ingestive behaviour also regulate reproductive function, the present study was designed to determine if central administration of MCH changes pulsatile secretion of luteinizing hormone (LH) in the rats. Wistar-Imamichi strain female rats were ovariectomized and implanted with oestradiol to produce a moderate inhibitory feedback effect on LH release. The effects of i. c.v. injections of MCH on LH release were examined in freely moving animals. Blood samples were collected every 6 min for 3 h through an indwelling cannula. After 1 h of sampling, MCH (0.1, 1 or 10 microg/animal) or vehicle (saline) was injected into the third cerebroventricle. Because MCH is also reported to affect the hypothalamo-pituitary-adrenal (HPA) axis, which in turn, can influence reproductive function, plasma corticosterone concentrations were determined in the same animals at 30-min intervals during the first and last hours and every 12 min during the second hour of the 3-h sampling period. When expressed as per cent changes, mean plasma LH concentrations after MCH administration were significantly lower in the animals injected with all doses of the peptide compared with vehicle-treated animals; LH pulse frequency was significantly lowered by 1 microg of MCH. Per cent changes in mean plasma corticosterone levels were not significantly affected by MCH administration. These results in oestradiol-treated ovariectomized rats indicate that central MCH is capable of inhibiting pulsatile LH secretion. We have previously shown that 48-h fasting suppresses pulsatile LH release in the presence of oestrogen. Take together, these results raise the possibility that MCH could play a role in mediating the suppression of LH secretion during periods of reduced nutrition.     

Endogenous Catecholamines Modulate Growth Hormone Release in the Conscious Rat During Hypoglycaemia but not in the Basal State

To investigate the role of endogenous catecholamines and 5-hydroxytryptamine in the control of growth hormone (GH) secretion, secretory profiles of GH and prolactin were measured in conscious, male rats following intravenous administration of either 1) α₂ antagonist idazoxan 2 mg/kg, a dose that blocked α₂ agonist induced GH rise, 2) α₁ antagonist prazosin 1 mg/kg, 3) non-specific β-blocker propranolol 1.5 mg/kg, a dose that prevented β₂ agonist (salbutamol) induced inhibition, 4) serotonin antagonist cyprohep-tadine 0.5 mg/kg, a dose that inhibited serotonin agonist quipazine induced GH rise, or 5) control. No drug altered mean GH or prolactin levels and pulsatile GH release persisted. Unilateral injections of prazosin, propranolol and idazoxan were made into the medial basal hypothalamus and preoptic-anterior hypothalamic area and of cyproheptadine into the medial basal hypothalamus, all with no effect on short-term GH release. GH and prolactin secretory profiles were measured after giving rats 6 units/kg intravenous insulin. Blood glucose levels fell to less than 50% basal. Hypoglycaemia caused a non-significant 30% fall in mean 2 h GH. Intravenous idazoxan, prazosin, propranolol and cyproheptadine (doses as in first study) did not modify the blood glucose fall, but idazoxan produced a significant reduction of mean GH compared to insulin alone (4 ± 1.1 ng/ml SEM, idazoxan/insulin versus 16 ± 5.6 ng/ml, saline/insulin). The lack of an effect of a- and β-blockers on normal, pulsatile GH release is against a role for endogenous catecholamines in controlling this release. Catecholamines may be released during insulin-induced hypoglycaemia and preferentially bind to α₂ receptors to increase GH secretion, thus attenuating the suppression of GH levels that would otherwise occur.     

Masculinizing effect of dihydrotestosterone on growth hormone secretion is inhibited in ovariectomized rats with anterolateral deafferentation of the medial basal hypothalamus or in intact female rats

There is a striking sex difference in the pattern of growth hormone (GH) secretion in rats. Our previous studies showed that short-term administration of pharmacological doses of testosterone or dihydrotestosterone (DHT) masculinized the GH secretory pattern in ovariectomized (OVX) rats. The locus where testosterone or DHT interacts with the somatotropic axis is believed to be the hypothalamus. To obtain insights into this phenomenon, we administered a single dose of DHT s.c. to adult OVX rats at 0.01, 0. 1 or 1 mg/rat. Blood GH concentrations were measured in unanaesthetized rats. Six to 12 h after the s.c. administration of all three doses of DHT, the GH secretory pattern revealed a male-like secretory pattern as shown by episodic bursts occurring at 2-3-h intervals with low or undetectable trough levels. When anterolateral deafferentation of the medial basal hypothalamus (ALC) was performed, the blood concentrations revealed irregularly occurring small fluctuations, instead of the usual high bursts, but the basal GH concentration was significantly higher than that of OVX-sham-operated rats. DHT treatment did not elicit pulsatile GH secretion or alter GH concentrations in OVX rats with ALC. When intact adult female rats received DHT at a dose of 1 mg/rat, the male-like GH secretory pattern was not induced. These results suggest that neural inputs from the anterolateral direction to the medial basal hypothalamus are necessary for the masculinizing effect of DHT on the GH secretory pattern in OVX rats, and that oestrogen in intact female rats prevents the masculinizing effect of DHT.