Intracerebroventricular injection of stresscopin-related peptide enhances cardiovascular function in conscious rats.

Stresscopin-related peptide (SRP), which is a member of the corticotropin-releasing factor (CRF) family, is a high-affinity ligand for the type 2 corticotropin-releasing factor receptor (CRF-R2) and is involved in stress-coping responses. Central treatment with SRP suppresses food intake, delays gastric emptying and decreases heat-induced edema, but the effects of central administration of SRP on the cardiovascular system are unclear. Here we examined the effects of intracerebroventricular (i.c.v.) administration of SRP on cardiovascular function, and compared the cardiovascular effects of SRP and stresscopin (SCP). Our results showed that i.c.v. administration of SRP (0.5nmol) increased mean arterial blood pressure (MABP) and heart rate (HR), but failed to increase plasma norepinephrine and epinephrine levels. Compared with an equivalent dose of SCP, the area under the curve (AUC) values for the changes in MABP and HR were significantly smaller with SRP, indicating that the cardiovascular effects of SRP were weaker than those mediated by SCP. Pre-treatment with a selective CRF-R2 antagonist, antisauvagine-30 (4nmol, i.c.v.) abolished the SRP and SCP induced changes in MABP and HR. These results indicate that central administration of SRP induces a weaker enhancement of cardiovascular function through CRF-R2 than that induced by SCP and that these effects are mediated without increasing plasma norepinephrine and epinephrine levels.

Effects of stresscopin on rat hypothalamic paraventricular nucleus neurons in vitro.

The effects of stresscopin (SCP) on rat paraventricular nucleus (PVN) neurons were examined using whole-cell patch-clamp recordings and single-cell reverse-transcription multiplex polymerase chain reaction (SC-RT-mPCR) techniques. Under current-clamp conditions, bath application of SCP (100 nM) induced inhibition in 35.2% (37/105) of putative magnocellular neurons and 24.7% (20/81) of putative parvocellular neurons, and excitation in 5.7% (6/105) of putative magnocellular neurons and 18.5% (15/81) of putative parvocellular neurons. SCP-induced inhibition persisted in the presence of a mixture of TTX, a voltage-gated Na+ channel blocker, CNQX, an AMPA/kainate receptor antagonist and bicuculline, a GABA(A) receptor antagonist, whereas SCP-induced excitation of PVN neurons was reversed by the mixture. The SCP-induced inhibition of PVN neurons was abolished by bath application of antisauvagine-30, a selective CRF receptor 2 (CRF-R2) antagonist. Under voltage-clamp conditions, SCP evoked outward currents at the holding potential (-60 mV), which reversed near the potassium equilibrium potential. The SCP-evoked membrane currents were completely blocked by bath application of tertiapin-Q, a selective blocker of G protein-activated inwardly rectifying potassium (GIRK) channels. SC-RT-mPCR analysis indicated that all the SCP-sensitive PVN neurons (57 SCP-inhibited neurons, 21 SCP-excited neurons) expressed CRF-R1 and CRF-R2 mRNAs. Among SCP-hyperpolarized PVN neurons, oxytocin (OT) mRNA was detected in 91.8% of putative magnocellular neurons and 45.0% of putative parvocellular neurons. OT mRNA was also detected in 26.6% of SCP-depolarized parvocellular neurons, but not in SCP-depolarized magnocellular neurons. These results indicate that SCP inhibits a subpopulation of PVN neurons, especially OTergic magnocellular neurons, by enhancing the activity of GIRK channels via CRF-R2.

Corticotrophin-releasing factor inhibits neuromedin U mRNA expressing neuron in the rat hypothalamic paraventricular nucleus in vitro.

In the present study, we examined the effects of corticotrophin-releasing factor (CRF) on neuromedin U (NMU) mRNA-expressing neurons in the rat paraventricular nucleus (PVN) by whole-cell patch-clamp recordings and single-cell reverse transcription-multiplex polymerase chain reaction (single-cell RT-mPCR) techniques. In total, of 116 PVN putative parvocellular neurons screened for NMU mRNA, 14.7% (17/116) of them expressed NMU mRNA. The electrophysiological properties observed in the NMU mRNA-expressing neurons were generation of a low-threshold Ca(2+) spike (LTS) and robust low voltage-activated (T-type) Ca(2+) currents. Under current-clamp conditions, CRF (100 nM) induced a reversible decrease in spike firing and significantly diminished the LTS in 88.2% (15/17) of NMU mRNA-expressing neurons. Extracellular application of 1 µM α-helical CRF-(9-14) (α-hCRF), a selective CRF receptor antagonist, completely blocked the CRF-induced decrease in spike firing in the NMU mRNA-expressing neurons. Under voltage-clamp conditions, CRF (100 nM) significantly decreased the peak value of the T-type Ca(2+) currents by 35.6±7.8%. These findings suggest that CRF decreases neuronal excitability and diminishes T-type Ca(2+) currents in a population of rat PVN NMU phenotype neurons in vitro.

Effect of hypertonic saline on rat hypothalamic paraventricular nucleus parvocellular neurons in vitro.

The effects of hypertonic saline on hypothalamic paraventricular nucleus (PVN) parvocellular neurons were examined using whole-cell patch-clamp technique. Under current-clamp, 50% (41/82) of parvocellular neurons were depolarized than the predicted values by hypertonic saline, and associated with increasing action potential frequency. Under voltage-clamp, unless hypertonic saline induced a shift of reverse potential to more positive values, neither mannitol nor hypertonic saline obviously increased the conductance in parvocellular neurons. Moreover, spontaneous excitatory postsynaptic currents (sEPSCs) were increased by isotonic increases in [Na(+)](o) in the parvocellular neurons. Bath application AMPA receptor antagonist CNQX or non-selective glutamate antagonist kynurenic acid almost completely blocked the sEPSCs. Extracellular application of gadolinium (Gd(3+)) blocked the hypertonic saline-induced response. These results suggested that subpopulation of PVN parvocellular neurons are selectively sensitive to NaCl. Hypertonic saline excited the PVN parvocellular neurons through Na(+)-detection and the excitatory glutamatergic synaptic input.

Involvement of vasopressin V1b receptor in anti-anxiety action of SSRI and SNRI in mice.

Arginine vasopressin (AVP) is critical in the regulation of hypothalamic-pituitary-adrenal axis activity, a major component of the stress response. The vasopressin V1b receptor (V1bR) mediates the stimulatory effect of AVP on adrenocorticotropin release. Previous studies showed that AVP facilitates aggression while serotonin inhibits aggression by blocking the activity of the vasopressin system. To examine whether the interaction of the V1bR and serotonin in the central nervous system controls anxiety-related behavior, we investigated the effects of acute and chronic treatment with a selective serotonin reuptake inhibitor (SSRI) and with a serotonin noradrenalin reuptake inhibitor (SNRI) on V1bR knockout (KO) mice and on V1bR antagonist (SSR149415)-treated mice. The effects were evaluated in experiments using an elevated plus-maze (EPM) test and a hole-board (HB) test, well established tests for evaluating anxiety-like behavior. For both the V1bR KO mice and V1bR antagonist-treated mice, acute treatment with either SSRI or SNRI did not change the time spent on the EPM open arms or the number of head dips in the HB. Chronic treatment of V1bR KO mice with SSRI did not change the amount of time spent on the open arms, the number of head dips, or the number of rearings, while chronic treatment with SNRI significantly increased the time spent on the open arms and the number of head dips. These results suggest that the anti-anxiety action of 5-HT reuptake inhibitors might partly involve V1bR regulating the anxiety behaviors.

Molecular regulation of antioxidant ability in the hippocampus of EL mice.

We recently found that the antioxidant ability was remarkably decreased in the hippocampus (Hipp) of EL at 8 weeks of age utilizing ESR spectroscopy. In this study, in addition to evaluating the extracellular glutamate concentration, we tried to determine whether or not changes in the expression of cystine/glutamate exchanger (xCT) and glutamate transporter take place in the Hipp of EL. EL mice and DDY mice at 5, 10, and 20 weeks of age were used for Exp. I and II, respectively. Exp. I: During the interictal state, dialysate was collected from the ventral Hipp using a microdialysis technique, and an extracellular concentration of glutamate ([Glu](o)) was measured with HPLC-ECD. Exp. II: The hippocampal expression of the glutamate transporter and xCT was estimated by Western blots. Exp. I: The level of [Glu](o) at 10 weeks of age was remarkably higher at other ages of EL mice, while [Glu](o) of DDY was unchanged as a result of age. Exp. II: The excitatory amino acid carrier-1 (EAAC-1) and xCT of EL mice at 10 weeks of age decreased more than those of DDY. GLAST and GLT-1 of EL mice at 5 weeks of age decreased more than those of DDY at the same age. No differences were found between EL and DDY for GLAST and GLT-1 at other ages. According to previous studies, the decreased endogenous antioxidant potential observed at 10 weeks of age is a very likely explanation for ictogenesis. The decreased xCT expression at 10 weeks of age could provide the molecular mechanism to explain the depletion of the endogenous antioxidant ability of EL mice during ictogenesis. In addition to the depletion of antioxidant ability, decreased EAAC-1 at this period could be one reason for the collapse of the molecular action of inhibition. These molecular findings support the idea that the elevation of [Glu](o) at 10 weeks of age triggers ictogenesis.

Different response between production of free radicals induced by central and peripheral administration of interleukin-1beta in conscious rats.

We examined whether central or peripheral administration of interleukin-1beta (IL-1beta) might change levels of nitric oxide (NO) and hydroxyl radical (*OH) in the medial prefrontal cortex (mPFC). Extracellular levels of NO metabolites (NOx(-)) and 2,3-dihydroxybenzoic acid (2,3-DHBA), as a marker of *OH production, were determined with an in vivo microdialysis technique in conscious rats. In the mPFC, central administration of IL-1beta into the mPFC resulted in dose-dependent increases in levels of both NOx(-) and 2,3-DHBA. In contrast, peripheral administration of IL-1beta significantly increased NOx(-) levels but not 2,3-DHBA levels. Perfusion of Mn(III) tetrakis (4-benzoic acid) porphyrin chloride, a superoxide (O(2)(-)) dismutase mimic, into the mPFC reduced the increases in levels of 2,3-DHBA induced by centrally administered IL-1beta, but enhanced the increases in levels of NOx(-) induced by centrally administered IL-1beta. The present results show a different response in free radical productions in the mPFC between central and peripheral administration of IL-1beta. This finding should be useful for our understanding of the response of NO and free radicals such as *OH and O(2)(-) in the mPFC after central and peripheral administration of IL-1beta.

The effects of centrally administered dexmedetomidine on cardiovascular and sympathetic function in conscious rats.

BACKGROUND: The alpha2-receptor is expressed in the brain, including the hypothalamus, where it is implicated in autonomic nervous system control. The effects of systemic administration of dexmedetomidine (DEX) on cardiovascular responses are well known; however, little is known about the effects of central administration of DEX on cardiovascular responses in conscious animals. In this study, we explored the effects and the mechanism of intracerebroventricularly (icv) administered DEX on cardiovascular responses and sympathetic nerve activity in conscious, unrestrained rats. METHODS: We administered DEX (0.5, 1, and 2 microg/kg) icv and measured the mean arterial blood pressure (MAP), heart rate (HR), and plasma catecholamine in conscious rats (n = 58). Rats were also administered atropine (n = 8), propranolol (n = 8), or hexamethonium (n = 8) to assess the influence of vagal or sympathetic efferent activity in the DEX-induced responses. Some of the rats underwent carotid sinus and aortic nerve denervation to exclude the effect of the baroreceptor reflex. RESULTS: Intracerebroventricular administration of DEX dose-dependently decreased MAP, HR, and plasma norepinephrine. Large dose of DEX decreased plasma epinephrine. The amplitude of MAP reduction induced by DEX was reduced by hexamethonium or propranolol. The amplitude of HR reduction was reduced by atropine or propranolol. The amplitude of MAP and HR reduction induced by DEX were smaller in hexamethonium-pretreatment rats than in intact ones. The amplitude of MAP and HR reduction induced by DEX were larger in sinus and aortic nerve denervation rats than in intact ones. CONCLUSIONS: These results indicate that icv administration of DEX decreases MAP by sympathetic inhibition and decreases HR by sympathetic inhibition and vagal stimulation.

Activation of a G protein-coupled inwardly rectifying K+ current and suppression of Ih contribute to dexmedetomidine-induced inhibition of rat hypothalamic paraventricular nucleus neurons.

BACKGROUND: Alpha2-adrenoceptor agonist has been reported to produce inhibition of arginine vasopressin release, diuresis, and sympatholytic effects. However, its mechanisms of central action remain incompletely understood. Hypothalamic paraventricular nucleus (PVN) neurons, which are in direct contact with noradrenergic synapses and are controlled by the hyperpolarization-activated currents, are called Ih (H current). The effect of dexmedetomidine, a highly selective and potent agonist, at alpha2 adrenoceptors on Ih is unknown. The purpose of this study was to examine the effects of dexmedetomidine on the PVN neuron, which is involved in the arginine vasopressin release and autonomic regulation. METHODS: The authors investigated the effects of dexmedetomidine on the membrane properties in PVN magnocellular neurons and an Ih in PVN parvocellular neurons with a whole cell patch clamp technique using a rat brain slice preparation. RESULTS: Dexmedetomidine dose-dependently hyperpolarized PVN magnocellular neurons. In the voltage clamp mode, dexmedetomidine induced an outward current, with a reversal potential of -94 mV, and this was shown to depend on the external concentration of K. Pretreatment with Ba or peptide toxin tertiapin blocked hyperpolarization induced by dexmedetomidine. The effect of dexmedetomidine was blocked by an alpha2-adrenoceptor antagonist, yohimbine. Ih was suppressed dose dependently by dexmedetomidine in PVN parvocellular neurons. Pretreatment with Cs occluded the Ih suppression by dexmedetomidine. Yohimbine blocked the Ih suppression by dexmedetomidine. The Ih sensitive to dexmedetomidine was weakly modulated by intracellular cyclic adenosine monophosphate. CONCLUSIONS: Dexmedetomidine inhibited PVN magnocellular neurons by activation of the G protein-coupled inwardly rectifying K current and inhibited PVN parvocellular neurons by suppression of Ih.

Age-dependent changes in the hippocampal antioxidant ability of EL mice.

Electron spin resonance (ESR) spectroscopy combined with in vivo microdialysis was used to analyze the antioxidant ability in the hippocampus of mice in an interictal state of EL mice utilizing decay ratio of an exogenously applied nitroxide radical (3-methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine-1-oxyl (PCAM)). In EL mice with a history of frequent seizures, the half-life of the electron paramagnetism of PCAM in the hippocampus was prolonged. These results revealed decreased antioxidant ability, suggesting vulnerability against oxidative stress. Our data suggest that epileptogenesis in EL mice with chronic seizures is associated with functional failure due to the oxidized redox state and revealed that the decreased hippocampal antioxidant ability is related to the regional vulnerability to oxidative stress in the limbic system of EL mice during epileptogenesis.

Effects of intracerebroventricular administration of neuropeptide W30 on neurons in the hypothalamic paraventricular nucleus in the conscious rat.

We demonstrated that intracerebroventricular (i.c.v.) administration of NPW30 increases the arterial blood pressure (ABP), heart rate (HR), and plasma catecholamine concentrations in conscious rats. NPW has been reported to be an important stress mediator in the central nervous system that modulates the hypothalamus-pituitary-adrenal (HPA) axis and sympathetic outflow. To examine the effects of NPW30 on the neural activity of the hypothalamic paraventricular nucleus (PVN), which is an integrative center of the autonomic and endocrine functions relevant to stress responses, we simultaneously recorded the single-unit activity in the PVN, ABP, and HR in conscious freely moving rats. Of the non-phasic (irregular) PVN neurons (n=35) examined, NPW30 (i.c.v. 3 nmol) elicited excitation in 22 neurons, inhibition in 7 neurons, and no response in 6 neurons, accompanied with increases in ABP and HR, whereas low-dose NPW30 (i.c.v. 0.3 nmol) did not affect the unit activity, ABP, or HR. Neurons that were affected by NPW30 were then further examined for their responses to perturbation in ABP and systemic administration of cholecystokinin-8 (CCK). The majority of neurons also showed responses to CCK, phenylephrine (PE), or nitroprusside (SNP). Our data suggest that central NPW30 modulates PVN neuronal activities, which might be involved in the regulation of cardiovascular function and energy balance through the autonomic nervous system, particularly, under stress-related conditions.

[Central action of orexins on sympathetic outflow and cardiovascular function with a focus on the paraventricular nucleus of the hypothalamus].

Orexins were initially reported as regulators of food intake. More recent reports suggest that they might play roles in the multiple functions of neuronal systems, causing medical conditions such as narcolepsy, a sleep disorder. Orexins and their receptors (OX1R and OX2R) are distributed in the neural tissue and brain regions involved in the autonomic and neuroendocrine control. Within the hypothalamus, orexin fibers and orexin receptors, especially OX2R, are found extensively in the paraventricular nucleus (PVN) of the hypothalamus. The PVN is an integrative center of the autonomic nervous system and the neuroendocrine system. Thus, orexins may play a role in the regulation of cardiovascular and autonomic nervous systems. This article provides a summary of our studies, in which we used direct recording of renal sympathetic nerve and PVN neuronal activities in conscious freely-moving rats and the in vitro whole cell patch-clamp technique to examine the direct effect of orexins on PVN neurons using a hypothalamic slice. Functional studies demonstrated that intracerebroventricularly (i.c.v.) administered orexins evoke increases in blood pressure, heart rate, and sympathetic nerve activity and depolarize both the magno- and parvo-cellular neurons through the activation of non-selective cation channels. The present studies suggest that PVN plays a role as one of the efferent pathways of orexin-induced activation of the sympathetic outflow.

Cardiovascular actions of central neuropeptide W in conscious rats.

Neuropeptide W (NPW) is a novel hypothalamic peptide that activates the orphan G protein-coupled receptors, GPR7 and GPR8. Two endogenous molecular forms of NPW that consist of 23- and 30-amino acid residues were identified. Intracerebroventricular (i.c.v.) administration of NPW is known to suppress spontaneous-feeding at dark-phase and fasting-induced food intake and to decrease body weight and plasma growth hormone and to increase prolactin and corticosterone; however, little is known about its effect on other physiological functions. We examined the effects of i.c.v. administration of NPW30 (0.3 and 3 nmol) on the mean arterial pressure (MAP), heart rate (HR), and plasma norepinephrine and epinephrine in conscious rats. NPW30 (3 nmol) provoked increases in MAP (85.12+/-3.16 to 106.26+/-2.66 mm Hg) and HR (305.75+/-13.76 to 428.45+/-26.82 beats/min) and plasma norepinephrine (138.1+/-18.1 to 297.2+/-25.9 pg/ml) and epinephrine (194.6+/-21.4 to 274.6+/-22.7 pg/ml). Intravenously administered NPW30 (3 nmol) had no significant effects on MAP and HR. These results indicate that central NPW30 increases sympathetic nervous outflow and affects cardiovascular function.

Enhanced cardiovascular alteration and Fos expression induced by central salt loading in a conscious rat transgenic for the metallothionein-vasopressin fusion gene.

The present study is an investigation of the responses of the cardiovascular system and Fos expression to intracerebroventricular (i.c.v.) administration of hypertonic saline (HS) in conscious arginine vasopressin (AVP)-overexpressing transgenic (Tg) and control rats. Central HS (0.3, 0.67, or 1.0M NaCl, 1 microl/min for 20 min) significantly increased the mean arterial blood pressure (MABP) and Fos-like immunoreactivity (FLI) in the paraventricular nucleus (PVN) and supraoptic nucleus (SON) of the hypothalamus, the area postrema (AP), the median preoptic nucleus (MnPO), and the organum vasculosum laminae terminalis (OVLT) in both Tg and control rats. The changes in MABP and FLI were significantly larger in Tg rats than in control rats. i.c.v. pretreatment with the AVP V1 receptor antagonist, OPC-21268, blocked the increase in MABP and significantly decreased the Fos expression in the PVN (posterior magnocellular (pm) component) induced by 0.3 M HS in the Tg rats. The present study demonstrates an increased responsiveness to i.c.v. administration of HS in AVP Tg rats, suggesting the relationship between the vasopressinergic drive and central cardiovascular response via, at least in part, the V1 receptor in the PVN magnocellular neurons.

Corticotrophin-releasing factor augments the I(H) in rat hypothalamic paraventricular nucleus parvocellular neurons in vitro.

The goal of this study was to characterize the effects of corticotrophin-releasing factor (CRF) on rat paraventricular nucleus (PVN) putative parvocellular neurons using whole cell patch-clamp recordings and single-cell reverse transcription-multiplex polymerase chain reaction (single-cell RT-mPCR) techniques. Under current clamp, CRF (10-600 nM) increased the neuronal basal firing rate and depolarized neurons in a dose-dependent manner. CRF-induced depolarization was unaffected by co-perfusion with TTX, 6-cyano-7-nitroquinoxaline-2 3-dione (CNQX), and bicuculline but was completely inhibited by ZD7288. Under voltage clamp, 300 nM CRF significantly increased the hyperpolarization-activated cation current (I(H)) in a voltage-dependent manner, shifted the I(H) conductance-voltage relationship (V 1/2) toward depolarization by approximately 7.8 mV, and enhanced the I(H) kinetics without changing the slope constant (k). Extracellular application of ZD7288 completely blocked I(H) and the CRF-induced increase in I(H). Furthermore, CRF-induced effects were completely blocked by extracellular application of 1 microM alpha-helical CRF-(9-14) (alpha-hCRF), a nonselective CRF receptor antagonist, but were not affected by extracellular application of antisauvagine-30, a selective CRF-receptor 2 antagonist. Single-cell RT-mPCR analysis showed that these neurons co-expressed CRF receptor 1 mRNA and CRF receptor 2 mRNA. Furthermore, CRF-sensitive neurons co-expressed HCN1 channel mRNA, HCN2 channel mRNA, and HCN3 channel mRNA, but not HCN4 channel mRNA. These results suggest that CRF modulates the subpopulation of PVN parvocellular neuronal function by CRF-receptor 1-mediated potentiation of HCN ion channel activity.

Neuromedin U receptor-2 mRNA and HCN channels mRNA expression in NMU-sensitive neurons in rat hypothalamic paraventricular nucleus.

We have characterized the neuromedin U (NMU)-sensitive neurons in the rat paraventricular nucleus (PVN) using whole-cell patch-clamp recordings and single-cell reverse transcription-multiplex polymerase chain reaction (single-cell RT-mPCR). Following completion of whole-cell recording, the NMU-sensitive neurons were examined for oxytocin (OT), vasopressin (VP), and corticotrophin-releasing hormone (CRH) mRNA expression using single-cell RT-mPCR. Of the NMU-sensitive neurons (n=23), 82% expressed OT mRNA, 9% expressed VP mRNA, 9% did not express the detected specific phenotypes mRNA. Further, the NMU-sensitive neurons (23/23) predominantly expressed NMU-receptor 2 (NMUR-2) mRNA, co-expressed HCN1 channel mRNA, HCN2 channel mRNA, and HCN3 channel mRNA but not HCN4 channel mRNA. These results suggest that NMU modulates the function of the PVN putative parvocellular neurons and is involved in the regulation of OTergic and VPergic neurons by enhanced HCN ion channels activity via NMU-receptor 2.

Regional differences in the expression of Fos-like immunoreactivity after central salt loading in conscious rats: modulation by endogenous vasopressin and role of the area postrema.

In this study, we examined the quantitative relationship between centrally administered hypertonic saline (HS) concentrations and the expression of Fos-like immunoreactivity (FLI) in brain regions involved in the homeostasis of body fluids. The regions examined were the organum vasculosum laminae terminalis (OVLT), the median preoptic nucleus (MnPO), the subfornical organ (SFO), the paraventricular nucleus (PVN), the supraoptic nucleus of the hypothalamus, the nucleus of the solitary tract (NTS), and the area postrema (AP). The experiments were performed in conscious rats with attention to the actual changes in central [Na(+)]. Hypertonic saline (0.3, 0.67, or 1.0 M) was delivered at 1 microl/min for 20 min. The changes in cerebrospinal fluid [Na(+)] during i.c.v. administration of 0.3 M hypertonic saline were compatible with those expected for thermal dehydration. FLI increased in a dose-dependent manner in the dorsomedial cap of the PVN and NTS. Although the pressor responses during central salt loading were not significantly affected by pretreatment with the peripheral vasopressin V(1) receptor antagonist OPC-21268, FLI expression in the PVN was significantly augmented. In addition, in AP-lesioned rats, FLI expression in the lateral magnocellular part of the PVN and NTS was significantly enhanced after central salt loading. These results suggest that the peripheral vasopressin system participates in negative feedback to modulate neuronal activities in the PVN, probably through the AP or direct action at the PVN in response to central osmotic and/or Na(+) stimulation.

Central stresscopin modulates cardiovascular function through the adrenal medulla in conscious rats.

Stresscopin (SCP or urocortin III), a member of the corticotropin-releasing factor (CRF) neuropeptide family, is a high-affinity ligand for the type 2 CRF receptor (CRF(2)). When administered peripherally, SCP suppresses food intake, delays gastric emptying and decreases heat-induced edema. Central administration of CRF produces marked hypertension and increased plasma catecholamine. However, the effects of SCP on the cardiovascular system are unknown. Thus, the present study compared the effects of intracerebroventricular (i.c.v.) administration of CRF and SCP on cardiovascular function. Central administration of SCP (0.05 or 0.5 nmol) elicited transient increases in mean arterial blood pressure (MABP) and heart rate (HR), and the higher dose of SCP (0.5 nmol) resulted in increased plasma epinephrine. In contrast, central administration of CRF provoked long-lasting increases in MABP, HR and plasma catecholamine levels (norepinephrine and epinephrine). Intravenously administered CRF and SCP (0.5 nmol) did not elicit significant changes in MABP and HR. Therefore, these data suggest that centrally administered SCP modulates cardiovascular function, likely through the sympatho-adrenal-medullary (SAM) system.

The alpha 1D-adrenergic receptor modulates cardiovascular and drinking responses to central salt loading in mice.

To characterize the involvement of specific alpha(1)-adrenergic receptor (alpha(1)-AR) subtypes in hypertension, parameters related to central salt- or angiotensin II (ANG II)-induced hypertension were investigated in alpha(1D)-AR-deficient mice (knockout). Baseline daily water intake and food intake were larger in alpha(1D)(-/-) mice than in alpha(1D)(+/+) mice. Intracerebroventricular (i.c.v.) administration of NaCl (0.67 M NaCl, 1 microl) elicited smaller increases in mean arterial blood pressure (MABP), heart rate, and water intake in alpha(1D)(-/-) mice than it did in alpha(1D)(+/+) mice. I.c.v. administration of ANG II (10 pmol) resulted in increases in MABP and water intake that were similar in alpha(1D)(-/-) mice and alpha(1D)(+/+) mice. These results suggest that alpha(1D)-AR is, at least in part, involved in central salt-induced but not ANG II-induced hypertension and water intake.

Effect of hypertonic saline on rat hypothalamic paraventricular nucleus magnocellular neurons in vitro.

The effect of hypertonic saline on rat hypothalamic paraventricular nucleus (PVN) magnocellular neurons was examined using a whole-cell patch-clamp technique. Under a current-clamp, 58/68 of magnocellular neurons were depolarized by hypertonic stimulation. Under a voltage-clamp, hypertonic saline produced an inward current via increased non-selective cationic conductance and shifting of the reversal potential to more positive values. Furthermore, hypertonic saline even without a change in osmolality increased spontaneous excitatory postsynaptic currents (sEPSCs). A bath application of CNQX almost completely blocked EPSCs. Extracellular application of gadolinium blocked the hypertonic saline- and mannitol-induced response. These results suggest that PVN magnocellular neurons are responsive to osmolality and Na+ concentrations. Hypertonic saline excited PVN magnocellular neurons via osmo-reception, Na+ -detection, and excitatory glutamatergic synaptic input.