Antagonism of ethanol-induced decrease in rat brain cGMP concentration by histidyl-proline diketopiperazine, a thyrotropin releasing hormone metabolite.

Intraperitoneal administration of thyrotropin releasing hormone (50 μmol/kg) produced an approximately 2-fold increase in rat brain cGMP concentration within 15 min. Histidyl-proline diketopiperazine, a metabolite of thyrotropin releasing hormone, produced a similar effect, but the response was faster and shorter-lasting. Intraperitoneal administration of ethanol (1.5 g/kg) decreased brain cGMP concentration approximately 50% within 10–15 min; thyrotropin releasing hormone or histidyl-proline diketopiperazine, injected 5 min after ethanol, antagonized the ethanol-induced decrease in cGMP. Antagonism of the ethanol-induced decrease in the cGMP level required 10 μmol/kg of thyrotropin releasing hormone but was observed with 5 μmol/kg of histidyl-proline diketopiperazine. These data suggest that the metabolic conversion of thyrotropin releasing hormone to histidylproline diketopiperazine might explain the previous observation that thyrotropin releasing hormone elevated the level of brain cGMP and antagonized the ethanolinduced decrease in brain cGMP concentration.     

The effects of thyrotropin releasing hormone and histidyl-proline diketopiperazine on delta-9-tetrahydrocannabinol-induced hypothermia

The effects of central and peripheral administration of thyrotropin releasing hormone (TRH) and its postulated metabolite, histidyl-proline diketopiperazine (HPD) on △9-tetrahydrocannabinol (THC) induced hypothermia in mice were investigated. Intraperitoneal administration of THC produced hypothermia. The peak response was observed between 1 and 2 hours and the hypothermia lasted for 5 to 6 hours. Intracerebral or intraperitoneal administration of TRH prior to THC injection antagonized the hypothermic response of the latter. Similar effects were produced by histidyl-proline diketopiperazine given intracerebrally. However, HPD was completely ineffective when given intraperitoneally. The antagonism of THC-induced hypothermia by TRH may be mediated by its conversion to HPD in the central nervous system.     

Muscimol induced feeding: A model to study the hypothalamic regulation of appetite

Intraventricular administration of the GABA agonist, muscimol, reliably induces feeding in sated rats in a dose dependent manner over a range from 50 to 500 ng. We used this pharmacological stimulant of appetite to examine the interrelationships of the peptides and monoamines involved in the hypothalamic regulation of appetite. Eating induced by muscimol (500 ng ICV) was suppressed by the opiate antagonist, naloxone; the dopamine antagonist, haloperidol; the cholinergic antagonist, atropine and by calcitonin. We could demonstrate no effect of thyrotropin releasing hormone or its metabolite histidyl-proline diketopiperazine or cholecystokinin-octapeptide, bombesin, isoproterenol, or phentolamine in doses known to suppress appetite. Based on these experimental results we propose a model of intrahypothalamic appetite regulation.     

Endogenous opiates inhibit gastric acid secretion induced by central administration of Thyrotropin-Releasing Hormone (TRH)

Thyrotropin-releasing hormone (TRH) administered intraventricularly (ICV) to rats causes a dose-dependent increase in gastric acid secretion over a range of 0.01 μg to 10 μg in the pyloris ligated rat. The maximum increase in gastric acid secretion occurs in the first hour. This effect of TRH is not mediated by its metabolites, histidyl-proline diketopiperazine or pyroglutamyl-histidyl-proline (acid TRH). β-endorphin, D-alanine-methionine-enkephalin and the leucine-enkephalin precursor, dynorphin, all inhibit TRH-induced gastric acid secretion. Bombesin, which reduces basal gastric acid secretion had no effect on TRH-induced secretion.     

Actions of intracerebroventricular administration of kyotorphin and an analog on thermoregulation in the mouse

Intracerebroventricular (ICV) administration of kyotorphin (L-Tyr-L-Arg) and cyclo (N-methyl-L-Tyr-L-Arg), its analog, produced significant dose-dependent hypothermic responses in mice at an ambient temperature of 24°C. The hypothermic action of kyotorphin was much greater than that of Met-enkephalin (Met-ENK) but less than that of cyclo NMTA. This action was slightly but not significantly reversed by intraperitoneally administered naloxone (8 mg/kg), an opioid receptor antagonist. Met-ENK utilized as a control peptide in this study also produced a dose-dependent hypothermia which was slightly antagonized by naloxone (8 mg/kg, IP). Thyrotropin releasing hormone (TRH) injected ICV produced hyperthermia dose-dependently. The hypothermia induced by kyotorphin, its cyclic analog and Met-ENK was prevented by a small dose of TRH (0.18 μg=0.5 nmol/animal) which by itself had little effect on body temperature. A TRH neuronal system in the brain may explain the mechanism of kyotorphin-induced hypothermia. However, there was little evidence of involvement of opioid receptors. The present study demonstrates a potent action of kyotorphin and its analog on thermoregulation.     

Modification of pentobarbital effects by natural and synthetic polypeptides: dissociation of brain and pituitary effects.

Thyrotropin releasing hormone (TRH) antagonizes pentobarbital sedation and hypothermia; somatotropin release inhibiting factor amplifies them. Other hypothalamic polypeptide releasing factors, Substance P and a variety of amino acids are ineffective. Three congeners of TRH antagonize pentobarbital; one amplifies it. The potencies of congeners as pentobarbital antagonists are unrelated to their potencies as pituitary thyrotropin releasers.     

Multiple expressions of the activity of guanine nucleotide regulatory protein in human thyroid adenylate cyclase

Adenylate cyclase (ATP pyrophosphate-lyase, EC 4.6.1.1) in plasma membranes from human thyroid was highly responsive to thyrotropin. Pretreatment of thyroid plasma membranes with 5′-guanylylimidodiphosphate (Gpp(NH)p) in the presence of Mg²⁺ led to a temperature-dependent activation, which was seen neither in the absence of Mg²⁺ nor at 4 °C. By contrast, thyrotropin bound to its receptors regardless of the temperature and produced its maximal effect after 2 min of preincubation in the absence or presence of Mg²⁺. Furthermore, activation was seen after treatment with thyrotropin and Gpp(NH)p even carried out in the absence of Mg²⁺ or at 4 °C. However, the full activation by Gpp(NH)p required Mg²⁺, hormone, and elevated temperature. These observations suggest that there appears to be two types of nucleotide interaction responsible for the Gpp(NH)p activation in human thyroid membrane; one type seen in the absence of hormone may represent the system uncoupled from hormone receptor, while the fully coupled hormone-sensitive adenylate cyclase accounts for the second type of interaction which requires the presence of hormone.     

Thyrotropin releasing hormone: direct evidence for stimulation of prolactin production by pituitary cells in culture

Addition of thyrotropin releasing hormone (TRH) to the medium of 2 clonal strains of functional rat pituitary cells stimulated the production of prolactin and inhibited growth hormone production. There was no effect on cell growth. Stimulation of prolactin production by TRH was detected within 4 hr, it reached a maximum level (2–5 times control) at 24–48 hr and persisted for at least 20 days in the continued presence of TRH. Stimulation was observed with a concentration of TRH as low as 0.10 ng/ml.     

Subcellular localization of alpha-melanocyte-stimulating hormone in the rat hypothalamus.

Homogenates of male rat hypothalami were fractionated by means of differential centrifugation, and α-melanocyte-stimulating hormone (α-MSH) in the various fractions was quantified by radioimmunoassay. Of the total quantity of α-MSH in the homogenate, 36% was recovered in the 11,500 g pellet and 31% sedimented between 11,500 and 105,000 g. α-MSH was not detected in the 105,000 g supernatant fluid. When the 900 g supernatant fluid was fractionated on continuous sucrose density gradients at non-equilibrium conditions, two populations of particles containing α-MSH were observed. When fractionated at equilibrium conditions, the two populations were recovered in a single band. These sedimentation characteristics indicate that the particles that contain α-MSH differ in size but are similar in density. After hypo-osmotic shock, the large particles containing α-MSH were not demonstrable, whereas the small particles appeared to be resistant to such treatment. In their sedimentation, the particles containing α-MSH were indistinguishable from particles containing thyrotropin releasing hormone (TRH) but were separable from those that contained luteinizing hormone releasing hormone (LHRH). It is suggested that the large particles containing α-MSH are synaptosomes.     

Unchanged response to stimulation of pituitary hormone release after serial UV irradiation in men

A group of 24 healthy young men were evaluated before and after serial suberythematous ultraviolet (UV) radiation: group I, control (no irradiation); groups II and III, 12 radiations in 4 weeks with two different spectra (both containing UV-B). Before the first and 2 days after the last exposure all the volunteers were given an intravenous injection of thyrotropin releasing hormone (TRH, protirelin 0.2 mg) and luteinizing hormone releasing hormone (LH-RH, gonadorelin 0.1 mg). The serum concentrations of TSH, follicle stimulating hormone, LH and prolactin were measured at 0, 20, 30, 45 and 60 min by radioimmunoassay. Neither basal nor stimulated levels of the pituitary hormones showed significant changes after UV radiation. The results showed that exposure to suberythematous doses of UV did not influence the regulation of pituitary hormones in these healthy individuals. Accepted: 24 October 1996     

The involvement of central cholinergic mechanisms in cardiovascular responses to intracerebroventricular and intravenous administration of thyrotropin-releasing hormone

Intracerebroventricular (i.c.v.) administration of thyrotropin-releasing hormone (TRH) in a range from 0.1 to 100 micrograms induced a dose-related increase in blood pressure in conscious rats, whereas TRH-free acid (TRH-OH) and histidyl-proline diketopiperazine (His-Pro-DKP), metabolites of TRH, did not. The blood pressure responses to intravenous (i.v.) injection of 5 mg/Kg TRH were similar to those induced by TRH (i.c.v.). Pretreatment with atropine (50 micrograms, i.c.v.) significantly reduced the pressor effect of TRH administered through either route. Hemicholinium-3 (50 micrograms, i.c.v.), an inhibitor of choline uptake, also prevented the increase in blood pressure induced by TRH (10 micrograms, i.c.v.). These results indicate that both centrally and peripherally administered TRH have pressor effects that are mediated by central cholinergic mechanisms, probably by activating cholinergic neurons.     

Effect of bombesin on basal and stimulated secretion of some pituitary hormones in humans

The effect of bombesin (5 ng/kg/min X 2.5 h) on basal pituitary secretion as well as on the response to thyrotropin releasing hormone (TRH; 200 micrograms) plus luteinizing hormone releasing hormone (LHRH; 100 micrograms) was studied in healthy male volunteers. The peptide did not change the basal level of growth hormone (GH), prolactin, thyroid-stimulating hormone (TSH), luteinizing hormone (LH) and follicle-stimulating hormone (FSH). On the contrary, the pituitary response to releasing hormones was modified by bombesin administration. When compared with control (saline) values, prolactin and TSH levels after TRH were lower during bombesin infusion, whereas LH and FSH levels after LHRH were higher. Thus bombesin affects in man, as in experimental animals, the secretion of some pituitary hormones.     

Hypothalamic hypopituitarism after hemiparesis.

Anterior pituitary insufficiency developed over 15 years in a man who had had a sudden episode of left hemiparesis and slurred speech at the age of 45 years. At age 60 radiographic studies revealed no evidence of a sellar or suprasellar neoplasm, and endocrinologic studies, including stimulation with thyrotropin releasing hormone and luteinizing hormone releasing hormone, yielded results consistent with hypopituitarism of hypothalamic origin. Response to thyroid, adrenocortical and testosterone therapy was good.     

Alteration of stimulated TSH and prolactin response in children treated with growth hormone.

The plasma TSH and prolactin responses to thyrotropin releasing hormone (TRH) were measured in 5 children with isolated growth hormone deficiency prior to, during and after the administration of human growth hormone (hGH). TSH and prolactin secretory patterns were not uniformly concordant. TSH responses to TRH infusion were suppressed in 4 subjects after 5 days or 1 month of hGH administration despite normal serum thyroxin concentrations. Prolactin responses were suppressed in all 5 subjects after 5 days of hGH administration. After 8 months of hGH therapy both TSH and prolactin responses returned toward pre-hGH values. Our finding that suppression of the TRH-induced TSH and prolactin secretory responses are reversible during hGH administration supports the concept of altered neuroregulation in this form of hypothalamic disorder.     

Neurotensin and bombesin: differential effects on body temperature of mice after intracisternal administration

Intracisternal administration of neurotensin or bombesin produces a significant hypothermic response in rodents in an ambient temperature of 23°C or below; bombesin has been reported to produce a significant hyperthermic response in rats at 36°C, but no change in colonic temperature at ambient temperatures between 31 and 33°C. In this study we compared the effects of the two neuropeptides on colonic temperature of mice exposed to different ambient temperatures to determine whether neurotensin also produces a poikilothermic state. From a series of experiments conducted at ambient temperatures of 4, 23, 26, 30, 34 and 38°C, in which mice received an intracisternal injection of an equimolar dose (0.6 nmol) of neurotensin or bombesin (or vehicle), we noted that the two neuropeptides produce different effects on colonic temperature. At ambient temperatures of 26°C and below, both neurotensin and bombesin produce a significant hypothermic response; however, at higher temperatures bombesin has no effect (30°C) or produces hyperthermia (34°C). In contrast, neurotensin produces hypothermia at 30°C and no significant effect at 34 and 38°C. In addition, a wide range of doses of neurotensin failed to produce the poikilothermic effects characteristic of centrally administered bombesin.     

Comparative studies of thyrotropin releasing hormone diazomethyl ketone and chloromethyl ketone analogs

Diazomethyl ketone and chloromethyl ketone analogs of thyrotropin releasing hormones have been synthesized and studied for their inhibitory effects on thyrotropin releasing hormone-induced release of radioactive ¹²⁵I-labelled hormones from the thyroid gland of eight-week old male Long-Evans rats. When Long-Evans rats were pretreated with thyrotropin releasing hormone diazomethyl ketone (TRH-DMK) or the chloromethyl ketone derivative (TRH-CMK), a dose-related inhibition of thyrotropin releasing hormone-induced ¹²⁵I release was observed which could be partially reversed by thyrotropin stimulating hormone (TSH). The diazomethyl ketone was a more effective inhibitor than the chloromethyl ketone. These compounds may act as an active-site directed antagonists whose effects are unique to the hypothalamo-pituitary-thyroid system.     

Episodic and TRH induced growth hormone release in primary hypothyroidism of man and rat.

In 27 hypothyroid subjects studied over 20 to 120 minutes, the concentration of serum growth hormone (GH) was variable with the amplitude and frequency of the secretory patterns similar to those reported by others for normal individuals. Serum GH, after the administration of thyrotropin releasing hormone (TRH) did not differ from values observed as spontaneous surges, in contrast to a consistent increase in thyrotropin and prolactin. Episodic secretion of GH persisted in thyroidectomized rats and did not differ significantly from that present in intact controls. It is concluded that episodic GH secretion is not abolished in primary hypothyroidism and that TRH is not a constant GH secretagogue in human subjects with hypothyroidism.     

Long term treatment with luteinising hormone releasing hormone agonists and maintenance of serum testosterone to castration concentrations.

Serum concentrations of luteinising hormone and testosterone were measured by radioimmunoassay one, two, four, seven, and 24 hours after the subcutaneous administration of 500 micrograms of the luteinising hormone releasing hormone agonist [D-Trp6, des-Gly-NH2(10)] LHRH ethylamide or [D-Ser(TBU)6, des-Gly-NH2(10)]LHRH ethylamide in patients who had previously received daily treatment with these peptides for 0, 1, 6, 12, 18, and 24 months. No increase in the serum concentrations of luteinising hormone or testosterone were detected at any time between one and 24 months'' treatment. The data show that daily subcutaneous administration of the two luteinising hormone releasing hormone agonists used at the appropriate dose can maintain concentrations of serum androgens equivalent to those after castration during long term treatment.     

Effects of bombesin on behavior

This report describes the influence of bombesin on the gross behavior of goldfish, frogs, mice, rats, guinea pigs, rabbits, chicks, pigeons and monkeys. Goldfish, frogs, chicks and pigeons were overtly unaffected by bombesin given centrally and/or peripherally. Mice, rats, guinea pigs, rabbits and monkeys responded quickly to intracerebroventricular (i.c.v.) and/or intrathecal (i.th.) administration of bombesin by displaying a range of behaviors suggestive of altered skin sensation. In mice, bombesin was essentially equipotent as a scratch inducer by i.c.v. and i.th. routes (A50 = 0.010-0.019 microgram) but 6800 times less potent i.p. In rats, bombesin-induced grooming and scratching behaviors were shown to be qualitatively different from those associated with ACTH-(1-24) and thyrotropin releasing hormone. Spantide and [D-Arg1, D-Pro2, D-Trp7,9, Leu11]substance P (both at 0.20, 0.50 and 0.80 microgram i.c.v.), two proposed bombesin receptor antagonists, did not markedly influence bombesin-induced scratching or hypothermia in rats.     

Changes in cardiovascular β-adrenoceptor responses during hypothermia

The purpose of this study was to determine cardiovascular β-adrenergic responses during hypothermia. In the present study, we used isoproterenol (Iso), a nonselective, potent β-adrenoceptor agonist, well known for its positive chronotropic and inotropic pharmacologic actions at normothermia. Rats were instrumented to measure mean arterial pressure (MAP) and left ventricular (LV) pressure–volume changes using a Millar pressure–volume conductance catheter. Core temperature was manipulated from 37 (normothermia) to 24 °C (hypothermia) and back to 37 °C (rewarming) using both internal and external heat exchangers. During cooling at each temperature (33, 30, 27, and 24 °C), central hemodynamic variables and MAP were measured while intravenously infusing Iso (doses of 1.7, 5, 10, and 20 ng/min). Seven animals underwent all phases of the protocol. At normothermia Iso infusion resulted in a significant, dose-dependent increase in heart rate (HR), stroke volume (SV), cardiac output (CO), LV dP/dt_max (left ventricular maximum derivative of systolic pressure over time) but no change in MAP. During cooling Iso infusion caused no dose-dependent change in any of the hemodynamic variables. After rewarming, baseline HR and LV dP/dt_max were increased, whereas SV was significantly reduced when compared with their pre-hypothermic baseline values. This study shows that physiological cardiovascular responses mediated by the β-adrenoceptor are significantly diminished during core hypothermia.