Lack of cardiovascular or ventilatory effects of melatonin in rats

The cardiovascular and ventilatory effects of centrally and peripherally administered melatonin were examined in both normotensive rats (NTR) and in spontaneously hypertensive rats (SHR). In the experiments on anaesthetised NTR melatonin was administered intravenously at doses of 1, 10, and 100 mumol/kg, or intracerebroventricularly at doses of 0.01, 0.1, 1, and 10 mumol/kg. In the experiments on conscious SHR melatonin was administered orally at doses of approximately 2 mg per animal per day, or intracerebroventricularly at doses of 0.01, 0.1, 1, and 10 mumol/kg. Melatonin did not produce any significant cardiovascular or ventilatory effects in any of the experiments.     

Intrathecal bombesin is sympathoexcitatory and pressor in rat

Bombesin, a 14 amino-acid peptide, is pressor when administered intravenously in rat and pressor and sympathoexcitatory when applied intracerebroventricularly. To determine the spinal effects of bombesin, the peptide was administered acutely in the intrathecal space at around thoracic spinal cord level six of urethane-anesthetized, paralyzed, and bilaterally vagotomized rats. Blood pressure, heart rate, splanchnic sympathetic nerve activity (sSNA), phrenic nerve activity, and end-tidal CO(2) were monitored to evaluate changes in the cardiorespiratory systems. Bombesin elicited a long-lasting excitation of sSNA associated with an increase in blood pressure and tachycardia. There was a mean increase in arterial blood pressure of 52 ± 5 mmHg (300 μM; P < 0.01). Heart rate and sSNA also increased by 40 ± 4 beats/min (P < 0.01) and 162 ± 33% (P < 0.01), respectively. Phrenic nerve amplitude (PNamp, 73 ± 8%, P < 0.01) and phrenic expiratory period (+0.16 ± 0.02 s, P < 0.05) increased following 300 μM bombesin. The gain of the sympathetic baroreflex increased from -2.8 ± 0.7 to -5.4 ± 0.9% (P < 0.01), whereas the sSNA range was increased by 99 ± 26% (P < 0.01). During hyperoxic hypercapnia (10% CO(2) in O(2), 90 s), bombesin potentiated the responses in heart rate (-25 ± 5 beats/min, P < 0.01) and sSNA (+136 ± 29%, P < 0.001) but reduced PNamp (from 58 ± 6 to 39 ± 7%, P < 0.05). Finally, ICI-216,140 (1 mM), an in vivo antagonist for the bombesin receptor 2, attenuated the effects of 300 μM bombesin on blood pressure (21 ± 7 mmHg, P < 0.01). We conclude that bombesin is sympathoexcitatory at thoracic spinal segments. The effect on phrenic nerve activity may the result of spinobulbar pathways and activation of local motoneuronal pools.     

Effect of porcine gastrin releasing peptide on anterior pituitary hormone release

The effect of porcine gastrin releasing peptide (GRP), a heptacosapeptide with potent gastrin releasing activity which has recently been isolated from porcine non-antral gastric tissue, on pituitary function was investigated in the rat. Graded doses of synthetic porcine GRP were injected intravenously and the animals were killed at various intervals after injection. Growth hormones, LH, FSH, and TSH were measured in serum by specific radioimmunoassays. GRP had no significant effect on growth hormone or FSH serum concentrations at any dose or sampling time studied. In contrast, the heptacosapeptide significantly stimulated LH and suppressed TSH secretion in a dose-related fashion. Since there are striking structural similarities between GRP and bombesin, a tetradecapeptide from amphibian skin which shows amino acid homology with the C-terminal region of GRP, GRP may be the mammalian counterpart of bombesin.     

The role of cholecystokinin octapeptide (CCK-8) in regulating thyrotropin secretion in rats. In vivo studies

The effect of cholecystokinin octapeptide (CCK-8) on basal and TRH-stimulated secretion of TSH was investigated in 67 male Sprague-Dawley rats. Blood for TSH determinations was sampled by cannulation of right heart auricle in urethane narcosis before and four times during 60 minutes following CCK-8 administration. It was found that CCK-8 administered to the lateral brain ventricle at a dose of 0.5 microgram per animal caused a decrease in blood serum TSH concentration but did not change the response of TSH to TRH. Intravenous administration of CCK-8 at doses of 2 and 20 micrograms/kg had no effect on blood serum concentration of TSH.     

Intravenous histamine reduces bombesin-stimulated gastrin release in dogs

The effect of histamine on gastrin release was studied in 7 conscious mongrel dogs with chronic gastric and duodenal fistulas. Histamine-2 HCl was infused in doses of 0 (control), 20, 40, 80, and 160 micrograms/kg per h for 2 h on separate days. During the second hour, bombesin 500 ng/kg per h was infused intravenously. Intragastric pH was constantly kept at 2.5 by intragastric titration during each test. Leakage of gastric contents into the duodenum was prevented by a prepyloric balloon passed retrograde through a duodenal fistula. Gastrin release, as expressed by the integrated response during the last 50 min of the bombesin infusion was significantly (P less than 0.05) decreased by all doses of histamine, compared to control. The infusion doses of histamine studied, 20, 40, 80, and 160 micrograms/kg per h reduced bombesin-stimulated gastrin release 16%, 19%, 19%, and 30%, respectively. This effect was blocked by a histamine H-2 but not an H-1 receptor antagonist. We conclude that by an H-2 mechanism, exogenous histamine reduces bombesin-stimulated gastrin release in dog.     

Oxytocin decreases plasma levels of thyroid-stimulating hormone and thyroid hormones in rats

Oxytocin (OXT) administered to rats induces several long-lasting physiological and metabolic effects. The aim of the present study was to investigate the effects of oxytocin treatment on plasma levels of thyroid-stimulating hormone (TSH), free thyroxine (fT4) and free triiodothyronine (fT3). For this purpose, oxytocin or NaCl was administered intracerebroventricularly (i.c.v.) (0.3 micro g) or subcutaneously (s.c.) (1 mg/kg) once a day for 5 days to male rats. Five or ten days after the last injection, rats were decapitated, blood was collected and hormone levels were analyzed by fluoroimmunoassay. The oxytocin treatment i.c.v. decreased plasma levels of TSH (p<0.05), fT3 (p<0.01) and fT4 (p<0.05) when measured at day 5 after oxytocin treatment, whereas the effect was abolished when measured at day 10. Oxytocin treatment s.c. did not affect plasma levels of TSH, fT3 or fT4. Thus, the effect seems to have been mediated within the central nervous system, and TSH and the thyroid hormones may be involved in some of the metabolic effects in response to oxytocin.     

Studies on bombesin-induced grooming in rats

The gross behavior induced by centrally administered bombesin in rats was compared to that elicited by ACTH-(1–24) and the somatostatin analog, des AA^{1,2,4,5,12,13}[D-Trp⁸]-somatostatin (ODT8-SS). Bombesin (0.001–1 μg, ICV) caused dose-related excessive scratching which was qualitatively different from that associated with the other two groom-inducing agents. Bombesin-induced grooming was not markedly affected by behaviorally nondepressant doses of haloperidol, morphine, naloxone or neurotensin. Bombesin was active in genetically hypotrichotic (essentially furless) rats; and, again in such animals, even after numbing the area caudal to the shoulders with lidocaine. Tolerance and cross-tolerance studies with bombesin and ODT8-SS indicated that they produce scratching through different mechanisms. Bombesin caused scratching when injected directly into the periaqueductal gray, but not when administered intravenously. Neither hypophysectomy nor adrenalectomy markedly affected bombesin-induced grooming. This behavior appears to be initiated in the central nervous system and is produced independently of the pituitary-adrenal axis.     

Bombesin-induced behavioural changes: Antagonism by neuroleptics

Adult male Sprague-Dawley rats were administered bombesin (BN) intracerebroventricularly (ICV), at a dose of 0.001, 0.01, 0.1, or 1.0 μg, and the behavioural effects monitored longitudinally across time for up to 24 hr. Administration of BN significantly increased the locomotor, rearing and grooming activity at all doses. The time-course of behavioural activation was dose-related (lasting up to 2.5 hr). There was no significant difference in the behavioural response of rats receiving the BN doses in an ascending or descending order. To test the effects of dopamine receptor blockade on the BN-induced behavioural changes, groups of animals were treated with fluphenazine or haloperidol (0.1 to 2.5 mg/kg, IP) 30 min prior to BN (1 μg, ICV) administration. The results revealed that the neuroleptics could effectively antagonize the BN-induced activation of locomotor, rearing and grooming activity. These data are concordant with the view that centrally administered BN stimulates spontaneous exploratory and grooming behaviours in rats, in a time- and dose-related manner. Furthermore, since neuroleptics block these effects, it remains possible that the BN-induced behavioural changes may be mediated, at least in part, through the dopaminergic system(s).     

The effect of beta-endorphin on basal and TRH-stimulated TSH release in conscious male rats

The inhibitory effect of beta-endorphin (EP) or other opioids on TSH secretion is, in contrast to their stimulating properties on PRL release, still a matter of debate. In the present study a dose of 1 microgram beta-EP injected intracerebroventricularly (IVT) in unstressed conscious male rats, though highly effective on PRL release, did not affect basal TSH levels, nor the TRH-induced TSH secretion. The previously reported inhibition of TSH release by opioids may therefore be an effect only seen when pharmacological doses are used.     

Combined use of dopamine and prostaglandin A1 in patients with acute renal failure and hepatorenal syndrome.

Dopamine and prostaglandin A1 were infused intravenously in 4 patients with the hepatorenal syndrome, in 1 patient with acute tubular necrosis, and 1 patient with cortical necrosis. Large doses of prostaglandin A1 decreased arterial blood pressure preventing increase in dosage; in contrast, high doses of dopamine elevated blood pressure. When the two drugs were administered conjointly, much larger doses of each agent could be administered without change in arterial blood pressure. Significant improvement of renal function was not observed in any of these critically ill patients during or within 24 hours after dopamine and prostaglandin A1 administration. This study demonstrated that extremely large doses of these vasodilating agents can be safely administered conjointly.     

Combined use of dopamine and prostaglandin A1 in patients with acute renal failure and hepatorenal syndrome

Dopamine and prostaglandin A₁ were infused intravenously in 4 patients with the hepatorenal syndrome, in 1 patient with acute tubular necrosis, and 1 patient with cortical necrosis. Large doses of prostaglandin A₁ decreased arterial blood pressure preventing increase in dosage; in contrast, high doses of dopamine elevated blood pressure. When the two drugs were administered conjointly, much larger doses of each agent could be administered without change in arterial blood pressure. Significant improvement of renal function was not observed in any of these critically ill patients during or within 24 hours after dopamine and prostaglandin A₁ administration. This study demonstrated that extremely large doses of these vasodilating agents can be safely administered conjointly.     

Stimulation and inhibition of food intake in sheep by centrally-administered hypothalamic releasing factors

Short-term effects of hypothalamic releasing factors on feeding behavior and digestive motility patterns were assessed in hay-fed sheep trained to eat more than half the total amount eaten over 8 h within the first 3 h after food presentation. Thyrotropin-releasing hormone (TRH) given intracerebroventricularly (ICV, 30 ng/kg) or intravenously at higher doses (IV, 3 micrograms/kg) reduced food consumption by 20 p. cent. The ICV or IV TRH-induced reduction was associated with behavioral excitation and stimulation of antroduodenal motor activity without changes in water intake. The ovine corticotropin releasing factor (oCRF 41) decreased food and water intake by 30-50% when administered ICV (60 ng/kg) but was not active when given systemically at doses up to 6 micrograms/kg. The synthetic human growth hormone releasing factor (hGRF 44) administered centrally (60 ng/kg) increased the amount of food intake and the antral motor activity without behavioral excitation. The results indicate a centrally-mediated facilitation of food intake by GRF and its inhibition by CRF which also affect water consumption. The presence of digestive motor effects suggests that extrapituitary pathways may be involved, as for TRH, in the action of both GRF and CRF.     

Dose-related involvement of CCK in bombesin-induced pancreatic growth.

We examined the role of CCK in bombesin-induced pancreatic growth in rats using the CCK receptor antagonist L-364,718. Rats (155 +/- 1 g, 8-10 per group) received subcutaneous injections every 8 h for 5 days with bombesin (0.6, 1.7 and 5 nmol/kg) or bombesin in combination with L-364,718 (1 mg/kg). After 5 days the pancreas was removed and pancreatic weight, protein content, DNA, amylase and chymotrypsin contents were determined. Bombesin produced a significant increase (48-475%) of pancreatic weight, tissue contents of protein, DNA, amylase and chymotrypsinogen (F = 82, P less than 0.001). When a large dose of bombesin (5 nmol/kg) was combined with L-364,718 a significant inhibition (up to 70%) of all tissue parameters was observed (P less than 0.001). L-364,718 did not affect the growth response to a small dose of bombesin (0.6 nmol/kg). Plasma CCK levels 15 min after a single injection of bombesin (0.6, 1.7 and 5 nmol/kg) were significantly increased in response to the 5 nmol/kg dose (2.0 +/- 0.7 to 3.4 +/- 0.8 pM, F = 6.9, P less than 0.01). No increases of CCK plasma levels were found in response to the 0.6 and 1.7 nmol/kg doses of bombesin, corresponding to the lack of effects of L-364,718 on growth parameters at these doses. Measuring the time-course of CCK plasma levels after a single injection of 5 nmol/kg bombesin revealed an increase from basal values of 1.4 +/- 0.3 pM to maximal levels of 3.5 +/- 0.5 pM after 15 min (F = 7.1, P less than 0.001). Values returned to basal after 60 min. These results suggest that low doses of bombesin act directly at the acinar cell or through release of non-CCK growth factors whereas high doses of bombesin act in part through CCK release.     

The effect of bombesin on basal, alpha-methyl-p-tyrosine, haloperidol, morphine, bremazocine and stress-induced prolactin secretion

Intravenously administered bombesin lowered basal PRL levels in conscious male rats and prevented the morphine, bremazocine and stress-induced PRL secretion. The same dose of bombesin had no effect on PRL levels in alpha-methyl-p-tyrosine pretreated rats and did not affect haloperidol-stimulated PRL release. These results show that bombesin given intravenously acts as an inhibitor of PRL secretion and suggests that it does not act on the lactotrope itself but rather by an increase of the inhibitory dopaminergic tone.     

Bombesin inhibits insulin release from isolated pancreatic islets of rats in vitro.

The effect of bombesin on insulin release from isolated pancreatic islets of rats was examined in vitro. Bombesin, at the doses ranging from 10 ng/ml to 1 microgram/ml, significantly inhibited 16.7 mM glucose-induced insulin release, while bombesin had no inhibitory effect on insulin release at 8.3 mM and 3.3 mM glucose. Moreover, bombesin also suppressed insulin release elicited by 10 mM arginine at the doses of 100 ng/ml and 1 microgram/ml. These results indicate that bombesin has a direct inhibitory action on insulin release.     

Further Observations on the Effect of Synthetic Thyrotrophin-releasing Hormone in Man

Synthetic thyrotrophin-releasing hormone (TRH) given intravenously in doses of 50 μg or more causes a significant rise in serum thyroid-stimulating hormone (TSH) levels but has no effect on serum growth hormone, plasma luteinizing hormone, or plasma 11-hydroxycorticosteroids under carefully controlled basal conditions.The peak TSH response to intravenous TRH occurs at 20 minutes. The mild and transient side effects, which occur only after intravenous TRH, include nausea, a flushing sensation, a desire to micturate, a peculiar taste, and tightness in the chest. There is considerable variability in response to a given dose of TRH in the same subject on different occasions and in different subjects. Oral administration of TRH in doses of 1 mg and above causes a rise in serum TSH, maximal at two hours, a consistent response being obtained at doses of 20 mg and above. A rise in serum protein-bound iodine (P.B.I.) follows that of TSH, a consistent response being observed at 40-mg doses of TRH orally. Measurements of serum TSH after intravenous administration of TRH or of serum TSH or serum P.B.I. after oral TRH should prove useful tests of pituitary TSH reserve.     

Effect of synthetic porcine gastrin-releasing peptide on plasma levels of immunoreactive cholecystokinin pancreatic polypeptide and gastrin in dogs

This study was conducted to determine if synthetic porcine gastrin-releasing peptide (GRP) stimulates the release of immunoreactive cholecystokinin (CCK), pancreatic polypeptide (PP) and gastrin in dogs. Three doses (0.01, 0.1 and 0.5 μg/kg-hr) of synthetic porcine GRP were administered intravenously to six conscious dogs. Synthetic procine GRP stimulated the release of each hormone in a dose-related manner. The effect of GRP on the response of gastrin was greater than its effect on CCK and PP responses. This study indicates that the biological action of synthetic porcine GRP is similar to the bombesin, an amphibian peptide shown previously to stimulate the release of gastrointestinal peptides.     

The effects of proglumide on cholecystokinin-, bombesin-, and glucagon-induced satiety in the rat

Intraperitoneal (IP) administration of the glutaramic acid derivative proglumide inhibited satiety induced by all IP doses of cholecystokinin octapeptide (CCK-OP) in 3-hour food-deprived intact rats. Proglumide did not influence satiety when administered alone and did not inhibit satiety induced by IP glucagon. While proglumide did not inhibit satiety induced by low doses of IP bombesin, it partially and significantly inhibited the satiety effects produced by high doses of this peptide. Since bombesin is a known secretagogue for CCK in several species, these results indicate that while bombesin and CCK act independently to induce satiety, the effect induced by high doses of bombesin is mediated, in part, by the release of endogenous CCK or a structurally related peptide. Furthermore, these results illustrate that proglumide is a specific antagonist of CCK-induced satiety and is, therefore, a potentially useful tool for investigating the physiologic role of this peptide in the control of food intake.     

Interactions of nimodipine and cocaine on endogenous catecholamines in the squirrel monkey

The effects of nimodipine on the cocaine-induced alterations in blood pressure, heart rate, and plasma catecholamines were studied in the squirrel monkey. Cocaine in intravenously administered doses of 0.5, 1, and 2 mg/kg produced significant increases in blood pressure and significant decreases in heart rate. These cardiovascular changes were associated with transient episodes of arrhythmias and with significant increases in plasma concentrations of dopamine, epinephrine, and norepinephrine. Nimodipine, 1 micrograms/kg/min for 5 min administered intravenously 5 min after cocaine, corrects the cardiovascular and plasma catecholamine concentration changes induced by this alkaloid. The same dose of nimodipine administered 5 min before cocaine prevents elevations of blood pressure. Plasma catecholamine increments are also prevented except for the highest dose of cocaine. Cardiovascular changes induced by cocaine administration in the squirrel monkey are temporally associated with significant increments in plasma catecholamines. Administration of nimodipine prevents or minimizes these endocrine and physiologic changes.