Takamitsu Hanamori

Central actions of adrenomedullin on cardiovascular parameters and sympathetic outflow in conscious rats

Adrenomedullin (ADM) is reported to be a peripherally acting hypotensive peptide, but its central actions are unclear. We investigated the effects of centrally administered ADM on blood pressure (BP), heart rate (HR), and renal sympathetic nerve activity (RSNA) in conscious rats and sinoaortic-denervated (SAD) rats. We also investigated the receptors interacting with ADM using two putative antagonists. Intracerebroventricular administration of ADM in doses of 0.1 and 0.5 nmol/kg caused tachycardia and early inhibition of RSNA. Central ADM (1.0 nmol/kg) induced hypertension, tachycardia, and a decrease followed by an increase in RSNA. In SAD rats, increases in BP, HR, and RSNA at the late phase were enhanced by central ADM (1.0 nmol/kg), whereas the early decrease in RSNA remained. Thus the inhibition of RSNA via central ADM may be unrelated to the arterial baroreceptor reflex. Pretreatment with antagonists human calcitonin gene-related peptide-(8-37) and human ADM-(22-52) significantly suppressed the central actions of ADM. The findings suggest that ADM is involved as a neuropeptide in the receptor-mediated central regulation of the cardiovascular system and RSNA.Adrenomedullin (ADM) is reported to be a peripherally acting hypotensive peptide, but its central actions are unclear. We investigated the effects of centrally administered ADM on blood pressure (BP), heart rate (HR), and renal sympathetic nerve activity (RSNA) in conscious rats and sinoaortic-denervated (SAD) rats. We also investigated the receptors interacting with ADM using two putative antagonists. Intracerebroventricular administration of ADM in doses of 0.1 and 0.5 nmol/kg caused tachycardia and early inhibition of RSNA. Central ADM (1.0 nmol/kg) induced hypertension, tachycardia, and a decrease followed by an increase in RSNA. In SAD rats, increases in BP, HR, and RSNA at the late phase were enhanced by central ADM (1.0 nmol/kg), whereas the early decrease in RSNA remained. Thus the inhibition of RSNA via central ADM may be unrelated to the arterial baroreceptor reflex. Pretreatment with antagonists human calcitonin gene-related peptide-(8-37) and human ADM-(22-52) significantly suppressed the central actions of ADM. The findings suggest that ADM is involved as a neuropeptide in the receptor-mediated central regulation of the cardiovascular system and RSNA.

Effects of area postrema lesion and abdominal vagotomy on interleukin-1β-induced norepinephrine release in the hypothalamic paraventricular nucleus region in the rat

Peripherally administered interleukin-1 beta (IL-1 beta) has been shown to increase extracellular norepinephrine (NE) concentration in the paraventricular nucleus (PVN) of the hypothalamus. The present study was carried out using an in vivo microdialysis technique in conscious rats in order to examine the possible involvement of the area postrema (AP) and the abdominal vagal afferent nerves in this effect. Extracellular NE concentrations in the PVN region were measured by high performance liquid chromatography with electrochemical detection. In AP-lesioned or abdominal-vagotomized rats, the NE increase was significantly attenuated compared to that in sham-operated rats; this reduction was greater in abdominal-vagotomized rats than in AP-lesioned rats. The results suggest that the AP as well as the abdominal vagal afferent nerves is involved in intraperitoneal (i.p.) administered IL-1 beta-induced NE release in the PVN region.

Convergence of afferent inputs from the chorda tympani, lingual-tonsillar and pharyngeal branches of the glossopharyngeal nerve, and superior laryngeal nerve on the neurons in the insular cortex in rats

The responses of single neurons in the insular cortex to electrical stimulation of the chorda tympani (CT), lingual-tonsillar branch of the glossopharyngeal (LT-IXth) nerve, pharyngeal branch of the glossopharyngeal (PH-IXth) nerve, and superior laryngeal (SL) nerve were recorded in anaesthetized and paralyzed rats. Ninety-four neurons responding to stimulation of at least one of the four nerves were identified from the insular cortex. Most of the neurons were located in the posterior portion of the insular cortex; the mean location was 0.8 mm anterior to the anterior edge of the joining of the anterior commissure (AC) and was 1.4 mm dorsal to the rhinal fissure (RF). Of the 94 neurons, 84 (89%) received convergent inputs from two or more nerves, and the remaining 10 (11%) received inputs from one nerve. The neurons responding to the CT stimulation were distributed more anteriorly than those responding to other three nerves in the anterior-posterior dimension. Our results indicate that the neurons recorded mainly from the posterior portion of the insular cortex receive convergent inputs from the oropharyngolaryngeal regions.

Cardiovascular and sympathetic effects of proadrenomedullin NH2-terminal 20 peptide in conscious rats.

Proadrenomedullin NH2-terminal 20 peptide (PAMP) and adrenomedullin (AM), which are derived from the same gene, are novel vasodilative peptides and have been shown to exhibit hypotensive action in anesthetized animals. To avoid the modification via anesthesia, we investigated the effects of intravenously administered PAMP on mean arterial pressure, heart rate (HR), and renal sympathetic nerve activity (RSNA) relative to those of AM in conscious unrestrained rats. We also examined whether the arterial baroreceptor reflex was altered with the two peptides. Intravenous injection of rat PAMP (rPAMP) (10, 20 and 50 nmol/kg) and rat AM (rAM) (0.3, 1.0 and 3.0 nmol/kg) similarly elicited dose-related hypotension accompanied by increases in HR and RSNA. However, the responses to rPAMP were less potent in magnitude and shorter in duration than those to rAM. Moreover, rAM facilitated baroreflex control, whereas rPAMP attenuated it. These findings indicate that although PAMP, as well as AM, may play an important role as a circulating hormone in the systemic circulation of conscious rats, the two peptides derived from an identical origin might have different mechanisms responsible for their cardiovascular and RSNA actions.

Abdominal vagotomy attenuates interleukin-1β-induced nitric oxide release in the paraventricular nucleus region in conscious rats

Nitric oxide (NO) has recently been shown to modulate the hypothalamic-pituitary-adrenal axis response to interleukin-1 beta (IL-1 beta). We measured levels of nitrite (NO2-) and nitrate (NO3-) in the hypothalamic paraventricular nucleus (PVN) region using an in vivo brain microdialysis technique in conscious rats. Intraperitoneally administered IL-1 beta produced a significant increase in both NO2- and NO3- levels in the PVN region. We also examined the possible involvement of the abdominal vagal afferent nerves in this effect. In abdominal-vagotomized rats, the increase was significantly attenuated compared to that in sham-operated rats. Our results suggest that the abdominal vagal afferent nerves are involved in intraperitoneally administered IL-1 beta-induced NO release in the PVN region.

Fiber types of the lingual branch of the trigeminal nerve, chorda tympani, lingual-tonsillar and pharyngeal branches of the glossopharyngeal nerve, and superior laryngeal nerve and their relation to the cardiovascular responses in rats

The effect of repetitive electrical stimulation at 50 Hz for 20 s of the lingual branch of the trigeminal nerve (LN), chorda tympani (CT), lingual-tonsillar (LT-IXth) and pharyngeal (PH-IXth) branches of the glossopharyngeal nerve, and superior laryngeal nerve (SLN) on the changes in arterial blood pressure (BP) and heart rate (HR) were investigated in anesthetized and paralyzed rats. The compound action potentials in these nerves were simultaneously recorded to know the relationships between the fiber types and the cardiovascular responses. In all nerves except the CT, repetitive electrical stimulation of the nerve elicited a tachycardia and an increase in BP. These cardiovascular responses were mainly related to the component-2 in the compound action potentials in respective nerves. The conduction velocities of the component-2 in the five nerves examined in the present experiment were between 9.5 and 17.0 m/s (mean, n = 4-7). Other components which have faster (component-1) or slower conduction velocities (component-3 and -4) than the component-2 were not likely to elicit the cardiovascular responses. These results suggest that nociceptive and taste fibers of A-delta fibers innervating the oral cavity and pharyngolaryngeal region largely contribute to the cardiovascular responses.

Fluctuations of the spontaneous discharge in the posterior insular cortex neurons are associated with changes in the cardiovascular system in rats

Extracellular neuronal responses were recorded from the posterior insular cortex. In three of 20 neurons, fluctuations in their spontaneous discharge were observed during recording without stimulation. In these recordings, fluctuations were also observed in arterial blood pressure (BP) and heart rate (HR) recorded simultaneously. For 3 neurons, the relationships among the fluctuations in the spontaneous discharge of the insular cortex neuron (INSneu), mean arterial pressure (MAP), and HR were analyzed using Pearsons correlation coefficient (r). In unit A, there was a negative correlation between INSneu and MAP (r = -0.30). The r between INSneu and HR was -0.02. In unit B, the data were divided into two groups according to HR (HR < 400 and HR > or = 400). The differential results in the correlations for INSneu-MAP were obtained in the two groups (r = -0.2, HR < 400; r = 0.51, HR > or = 400). For unit C, the INSneu was positively correlated with BP (r = 0.31) and HR (r = 0.36). From the correlation analysis concerning the time, changes in INSneu seem to precede (or delay) changes in BP. These results showed that fluctuations in neuronal activity in the posterior insular cortex are positively or negatively correlated with BP (or HR). The data suggest that some of the neurons in the posterior insular cortex may play a role in the homeostatic (and/or regulatory) control of the autonomic system.

Effects of electrical and chemical stimulation of the amygdala on the spontaneous discharge in the insular cortex in rats

Both the amygdala (especially, the basolateral nucleus of the amygdala) and the insular cortex are important for conditioned taste aversion. From the anatomical view point, there are reciprocal connections between the insular cortex and the amygdala. In the present study, we investigated the effect of electrical and chemical stimulation of the amygdala on the spontaneous discharge of the insular cortex neurons in anesthetized rats. In most neurons (10 of 14), spontaneous discharge was decreased after a microinjection of glutamate (Glu). In these neurons, the injection site was within the basolateral nucleus of the amygdala (the basolateral/lateral nuclei). On the other hand, when a gamma amino-butyric acid (GABA) was microinjected into the basolateral nucleus of the amygdala, none of the 5 neurons showed any change in spontaneous discharge. Electrical train stimulation of the basolateral nucleus of the amygdala (100 Hz, 2-6 s) depressed the spontaneous discharge of the neurons in the insular cortex, as in the case of a Glu microinjection. These results indicate that activation of the basolateral nucleus of the amygdala could depress the neuronal activity in the insular cortex. Such results may yield data leading to the elucidation of the neuronal mechanisms of conditioned taste aversion.

Effects of Systemic Interleukin-1β Administration on Daily Drinking and Renal Excretory Function in Conscious Rats

To elucidate the roles of interleukin-1beta (IL-1beta), a cytokine with several diverse actions, in the control of body fluid balance, its effects on daily drinking behavior and renal excretory function were examined in conscious rats. Administration of IL-1beta (4 microg/kg, I.P.) resulted in the suppression of both daily drinking and food intake and a decrease in daily urinary sodium and potassium excretion, but had no effect on urine volume. The IL-1beta-induced decrease in sodium excretion was abolished in renal-denervated rats. Kainic acid was then injected into the anteroventral third ventricle region, including the organum vasculosum of the lamina terminalis, to examine whether neurons in this region are involved in the IL-1beta-induced responses; the effects on daily drinking and urinary sodium and potassium excretion were abolished, whereas the effects on food intake, although attenuated, were still present. In contrast, electrical lesion of the subfomical organ did not affect the IL-1beta-induced responses. Thus, IL-1beta seems exert its effects on body fluid balance at several distinct sites in the central nervous system.

Difference in water intake but not in renal sympathetic nerve activity in response to central salt-loading or angiotensin II in awake Dahl salt-sensitive and -resistant rats

Experiments were conducted to examine whether renal sympathetic nerve activity (RSNA) and water intake in response to central salt-loading or angiotensin II (A II) differ between freely-moving Dahl salt-sensitive (DS) and -resistant (DR) rats maintained on a low-salt diet. Intracerebroventricular (i.c.v.) administration of hypertonic saline (0.3 M, 1 microl/min, 20 min) or A II (100 ng/1 microl) evoked water intake, pressor response and suppression of RSNA in both strains. The cumulative water intake in DS rats over a 60-min period after i.c.v. infusion of hypertonic saline or A II was significantly attenuated compared with that in DR rats. The RSNA response did not show a significant difference between the strains. These results demonstrate that water intake, but not RSNA response to acute central salt-loading or A II differ between awake DR and DS rats.