Autonomic Neuroscience: Basic & Clinical

Published by Elsevier
Online ISSN: 1566-0702
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Article
Previously, we found that the nitrate synthesized from glycerin, 2-nitrate-1,3-dibuthoxypropan (NDBP), increased NO levels in rat aortic smooth muscle cells, inducing vasorelaxation in mesenteric artery. However, its effects on blood pressure and heart rate as well as on autonomic function were not investigated. This study evaluated the action of NDBP on these cardiovascular parameters in spontaneously hypertensive (SHR) and Wistar Kyoto (WKY) rats. We found that NDBP causes a biphasic response: hypotension and bradycardia followed by hypertension and tachycardia in WKY and SHR rats. Atropine (2mg/kg) blunted the hypotension induced by NDBP (15mg/kg) in WKY and SHR (-75±9 vs -12±3mmHg, n=6; -101±6 vs -7±2bpm, n=6; respectively, p<0.05) and the pressor response to the compound was potentiated. Furthermore, vagotomy reduced the bradycardia in WKY and SHR (-136±8 vs -17±2, n=4, p<0.05; -141±9 vs -8±2, n=6, p<0.05). Moreover, hexamethonium (30mg/kg) reduced both bradycardia (-278±23 vs -48±3 in WKY; -285±16 vs -27±19 in SHR, n=4; p<0.05) and pressor response (28±8 vs -9±5-WKY, n=6; 42±7 vs -19±8-SHR, n=5; p<0.05). In addition, administration of methylene blue (4mg/kg) attenuated the hypotensive and bradycardic responses to the NDBP in all groups. In conclusion, NDBP induces bradycardia by direct vagal stimulation and pressor response by increasing sympathetic outflow to the periphery.
 
Article
We investigated how agonists at purinoceptors may affect tumour cell metabolism. This was investigated in vitro in tumour cell lines by microphysiometry, which method monitors extracellular acidification rate (ECAR), on-line. The cell lines investigated were the murine sarcoma, MCG 101, and the human colon cancer, HT-29. In MCG 101, adenosine-5'-triphosphate (ATP) or uridine-5'-triphosphate (UTP) caused a concentration-dependent increase in ECAR, most likely due to the ligation of P2Y(2) receptors, which response was blocked by suramin. In HT-29, ATP or UTP elicited a concentration-dependent, biphasic change in ECAR (increase/decrease). The pharmacological analysis suggests the involvement of P2Y(2) receptors, although other P2 receptor subtypes cannot be entirely excluded. This biphasic response to UTP or ATP was resistant to suramin. The expression of P2Y(2) receptors was demonstrated in both cell lines by immunocytochemistry and Western blot. The current study, thus, shows the functional and morphological expression of a purinoceptor subtype with partly different effects on metabolism in two different tumour cell lines.
 
Article
In light of the nonequilibrium thermodynamics by I. Prigogine, the autonomic nervous system as a whole may be viewed as a dissipative structure progressively assembled in the course of evolution, plastically and rhythmically interfaced between forebrain, internal and external environments, to regulate energy, matter and information exchanges. In the present paper, this hypothesis is further pursued to verify whether the two main divisions of the autonomic nervous system, the sympathetic and parasympathetic systems, may support different types of exchange with the external environment. Previous data from hypothalamic stimulation experiments, studies of locus coeruleus function and available data on behavioral functional organization indicate that (1) tight engagement with the external environment, (2) high level of energy mobilization and utilization and (3) information mainly related to exteroceptive sensory stimulation characterize a behavioral prevalence of sympathoadrenal activation. On the other hand, (1) disengagement from the external environment, (2) low levels of internal energy and (3) dominance of proprioceptive information characterize a behavioral prevalence of vagal tone. Behavioral matter exchanges such as feeding, drinking, micturition and defecation are equally absent at the extreme of sympathoadrenal and vagally driven behaviors. The autonomic nervous system as a whole is genetically determined, but the sympathoadrenal system has been mainly designed to organize the visceral apparatus for an action to be performed by the biological system in the external environment and to deal with the novelty of task and of the environment, while the functional role of the parasympathetic is to prepare the visceral apparatus for an action to be performed by the biological system on itself, for recovery and self-protection (homeostasis), and is reinforced by repetition of phylo- and ontogenetically determined patterns. The available clinical data further support this interpretation indicating that an increased sympathetic and a decreased vagal tone may represent a consistent risk factor for cardiovascular diseases.
 
Article
The role of the autonomic nervous system, the central and peripheral chemoreceptors, and the arterial baroreceptors was examined in the cardiovascular response to hypercapnia in conscious rats chronically instrumented for the measurement of arterial blood pressure (ABP), heart rate (HR), and renal sympathetic nerve activity (RSNA). Rats were exposed to hypercapnia (6% CO2), and the cardiovascular and autonomic nervous responses in intact and carotid chemo- and/or aortic denervated rats were compared. In intact and carotid chemo-denervated rats, hypercapnia induced significant increases in mean ABP (MABP) and RSNA, and a significant decrease in HR. The HR decrease was reversed by atropine and eliminated by bilateral aortic denervation, which procedure, however, did not affect the MABP or RSNA response. Bilateral carotid chemo-denervation did not affect the baroreflex control of HR, although this control was attenuated by aortic denervation. Hypercapnia did not affect baroreflex sensitivity in intact rats. These results suggest that hypercapnia induces an increase in MABP due to an activation of sympathetic nervous system via central chemoreceptors and a decrease in HR due to a secondary reflex activation of the parasympathetic nervous system via arterial baroreceptors in response to the rise in ABP. In addition, carotid chemoreceptors do not play a major role in the overall cardiovascular response to hypercapnia in conscious rats. The mechanism responsible for the parasympatho-excitation may also involve CO2 induced aortic chemoreceptor simulation.
 
Article
Our daily life not only involves calm, resting states but is filled with perturbations that induce active conditions, such as movements, eating, and communicating. During such active periods, cardiorespiratory regulation must be adjusted for bodily demands, which differ from those during resting states, by modulating or resetting baseline levels. To explore neural mechanisms of state-dependent adjustments of central autonomic regulation, we recently focused on the following two states: 1), stress-induced defense (fight-or-flight) responses, because stressors induce both cognitive, emotional, and behavioral changes and autonomic alterations, and 2), sleep/wake differences. Basal respiration and respiratory reflex regulation significantly differ during waking and sleep states. In this review, we will summarize our recent findings with orexin knockout and orexin neuron-ablated mice to determine possible contributions of orexin, a hypothalamic neuropeptide, to state-dependent adjustments of central autonomic regulation. The diversity of synaptic control of cardiovascular and respiratory neurons appears to be necessary for animals to adapt to ever-changing life circumstances and behavioral states. The orexin system likely functions as one essential modulator for coordinating circuits controlling autonomic functions and behaviors.
 
Article
The term 'metanephrines' is used to indicate the two catechol 3-O-methylated metabolites of epinephrine (E) and norepinephrine (NE): metanephrine and normetanephrine (NMN). The corresponding 3-O-methylated metabolite of dopamine is usually referred to as 3-methoxytyramine rather than 3-methoxydopamine and is not generally considered a "metanephrine". O-Methylation occurs outside the sympathetic neuron and neuroeffector junction. Metanephrines are products of the enzyme catechol-O-methyltransferase (COMT). Subsequent conjugation with sulfate or deamination by monoamine oxidase (MAO) followed by reduction to vanilmandelic acid (VMA) facilitates urinary excretion. For the clinician, measurement of normetanephrine provides an index of norepinephrine released during sympathetic nervous system activity, whereas metanephrine concentration provides an indication of adrenal medullary metabolism of epinephrine prior to its discharge into the circulation. Plasma epinephrine concentration is the preferable index of adrenal medullary epinephrine discharge. Pheochromocytomas, with their protean clinical manifestations, may be diagnostic challenges, but assay of metanephrines, especially plasma metanephrine, can be particularly helpful in diagnosis. These COMT metabolites may also help in elucidation of still undiscovered genetic and acquired disorders of catecholamine metabolism.
 
Article
The insular cortex interacts with the bulbar autonomic nuclei providing autonomic manifestations accompanying several neurological and psychosomatic disorders. The aim of our study was to identify the sites within the insular cortex, which could be responsible for the gastrointestinal, respiratory and cardiovascular responses. The main methods used were microinjections of HRP into several parts of the bulbar dorsal vagal complex and microstimulation of the insular cortex. It has been found that a compact group of neurons located in the middle level of the rat insular cortex projects directly to the specific "gastric" part of the dorsal vagal complex. Retrograde labelled cell bodies were revealed in the V layer of the disgranular and agranular insular cortex. Microstimulation of the sites within the middle level of the rat insular cortex produced gastric motor responses and a decrease in inspiratory airflow without significant alteration in respiratory cycle duration. More caudal microstimulation produced an increase in respiratory airflow and decreased respiratory cycle duration. These responses were usually accompanied by changes in the level of blood pressure. It is concluded that autonomic representation in the rat insular cortex is organised in a viscerotopic manner. The inhibitory respiratory zone overlaps with the gastrointestinal control area in the middle part of the insular cortex. More caudally, the excitatory respiratory zone overlaps with the cardiovascular area. On the basis of these experimental results and the data of others authors the general scheme of autonomic representation in the rat insular cortex is discussed.
 
Article
Chronic intermittent stimulation of the vagus nerve (VNS) is an approved adjunctive therapy of refractory epilepsy. Nevertheless, the circuits triggered by VNS under the variable conditions used in patients are not well understood. We analyzed the effect of increasing pulse frequency on physiological variables (intragastric pressure, cardiac and respiratory frequencies) and neuronal activation in the solitary tract nucleus (NTS), the entry level of peripheral vagal afferents, in the rat. For this purpose, we compared the subnuclear distribution of Fos-like immunoreactivity within the NTS following VNS at frequencies selected for their low (1 Hz) or high (10 Hz) therapeutic efficacy. In addition, NADPH diaphorase histochemistry was conducted in double-labeling experiments to check whether activated neurons may express nitric oxide (NO). We demonstrated that increasing pulse frequency had a major influence on the cardiorespiratory response to VNS and on the amount of activated neurons within NTS subdivisions engaged in cardiorespiratory control. These data, in line with clinical observations, suggested that within the range of therapeutic frequency, VNS may favor the regulation by vagal inputs of cortical activities within limbic areas involved in both epileptogenesis and cardiorespiratory afferent control. Furthermore, we did not find any evidence that anticonvulsant VNS might trigger NOergic neurons in the NTS.
 
Article
Severe cardiac autonomic denervation occurs in the acute Chagas' disease in rats. The present study aims at verifying whether this denervation was accompanied by impairment of heart function. Scorpionic (Tityus serrulatus) crude venom was used for neurotransmitter release in isolated hearts (Langendorff's preparation). In control hearts, the venom induced significant bradycardia followed by tachycardia. In infected animals, despite the severe (sympathetic) or moderate (parasympathetic) cardiac denervation, the venom provoked similar bradycardia but the tachycardia was higher. The hearts of infected animals beat at significantly lower rate. Atropine prevented this lower rate. Our results demonstrated sympathetic dysfunction during the acute phase of Trypanosoma cruzi infection in rats, the parasympathetic function being spared.
 
Article
Space motion sickness is experienced by 60% to 80% of space travelers during their first 2 to 3 days in microgravity and by a similar proportion during their first few days after return to Earth. Space motion sickness symptoms are similar to those in other forms of motion sickness; they include: pallor, increased body warmth, cold sweating, malaise, loss of appetite, nausea, fatigue, vomiting, and anorexia. These are important because they may affect the operational performance of astronauts. Two hypotheses have been proposed to explain space motion sickness: the fluid shift hypothesis and the sensory conflict hypothesis. The fluid shift hypothesis suggests that space motion sickness results from the cranial shifting of body fluids resulting from the loss of hydrostatic pressure gradients in the lower body when entering microgravity. The cranial fluid shifts lead to visible puffiness in the face, and are thought to increase the intracranial pressure, the cerebrospinal-fluid pressure or the inner ear fluid pressures, altering the response properties of the vestibular receptors and inducing space motion sickness. The sensory conflict hypothesis suggests that loss of tilt-related otolith signals upon entry into microgravity causes a conflict between actual and anticipated signals from sense organs subserving spatial orientation. Such sensory conflicts are thought to induce motion sickness in other environments. Space motion sickness is usually treated using pharmaceuticals, most of which have undesirable side effects. Further studies elucidating the underlying mechanism for space motion sickness may be required for developing new treatments.
 
Article
Apelin peptides are now known to be endogenous ligands at the orphan G-protein coupled receptor, APJ. Apelin and its receptor have been found in the brainstem and shown to have a role in haemodynamic homeostasis when injected intravenously. The physiological role of this peptide and its receptor centrally are yet to be elucidated. In this study, urethane anaesthetised, paralysed and ventilated male Sprague-Dawley rats (350-450 g, n=4) were used to investigate the action of apelin-13 microinjected directly into the nucleus tractus solitarius (NTS) and the rostral ventrolateral medulla (RVLM) on arterial pressure and phrenic nerve activity. Apelin-13 microinjections (4 mmol/l, 50 nl) into the NTS resulted in either apnea or decreased phrenic nerve discharge amplitude by up to 30%. In the RVLM, apelin-13 caused either a 100-200% increase, or a 10-30% increase in phrenic nerve discharge amplitude depending on the exact site of injection. Increases of 10-20 mm Hg in arterial pressure were also evoked from both the NTS and the RVLM. These data suggest a role for apelin-13 in arterial pressure and respiratory control in the NTS and the RVLM.
 
Article
A 14-day, 6 degrees head-down bed rest (HDBR) study was conducted with 12 healthy young men to determine whether there are transient responses of the cardiovascular autonomic regulatory system including cardiovascular, autonomic nervous, and cardiac baroreceptor reflex functions in the acute phases of HDBR and post-HDBR. Compared with the supine position before bed rest, the high-frequency band power (HF(RRI)) of RR intervals (RRIs) decreased significantly at 3, 6, and 24 h of HDBR. This tendency went on until 24 h post-HDBR. Three kinds of cardiac baroreceptor reflex sensitivity (BRS) were estimated from closed-loop approaches to simultaneously recorded spontaneous RRI and systolic arterial pressure (SAP) fluctuations. BRSsequence is based on the simultaneous changes between RRI and SAP. alphaLF and alphaHF are based on a cross-spectrum analysis for low- and high-frequency bands of RRI and SAP. Although BRSsequence decreased significantly at acute phases of both HDBR and post-HDBR, neither alphaLF nor alphaHF decreased significantly at any of the acute phases of HDBR and post-HDBR. Our results suggest that HF(RRI) and BRSsequence can be used effectively to reveal reductions in cardiac vagal nervous modulation on the sinus node and cardiac BRS within 24 h of both HDBR and post-HDBR.
 
Article
CNI-1493 is a potent anti-inflammatory agent, which deactivates macrophages and inhibits the synthesis of proinflammatory mediators. The objective of the present study was to identify the role of the central nervous system (CNS) and efferent vagus nerve signaling in CNI-1493-mediated modulation of acute inflammation in the periphery. CNI-1493 was administered either intracerebroventricularly (i.c.v., 0.1-1,000 ng/kg) or intravenously (i.v., 5 mg/kg) in anesthetized rats subjected to a standard model of acute inflammation (subcutaneous (s.c.) injection of carrageenan). I.c.v. CNI-1493 significantly suppressed carrageenan-induced paw edema, even in doses at least 6-logs lower than those required for a systemic effect. Bilateral cervical vagotomy or atropine blockade (1 mg/kg/h) abrogated the anti-inflammatory effects of CNI-1493 (1 microg/kg, i.c.v. or 5 mg/kg, i.v.), indicating that the intact vagus nerve is required for CNI-1493 activity. Recording of the efferent vagus nerve activity revealed an increase in discharge rate starting at 3-4 min after CNI-1493 administration (5 mg/kg, i.v.) and lasting for 10-14 min (control activity=87+/-5.4 impulses/s versus CNI-1493-induced activity= 229+/-6.7 impulses/s). Modulation of efferent vagus nerve activity by electrical stimulation (5 V, 2 ms, 1 Hz) of the transected peripheral vagus nerve for 20 min (10 min before carrageenan administration and 10 min after) also prevented the development of acute inflammation. Local administration of the vagus nerve neurotransmitter, acetylcholine (4 microg/kg, s.c.), or cholinergic agonists into the site of carrageenan-injection also inhibited acute inflammation. These results now identify a previously unrecognized role of efferent vagus nerve activity in mediating the central action of an anti-inflammatory agent.
 
Article
We studied autonomic functions in orexin-deficient mice and found abnormalities in the emotional state-dependent adjustment of the central autonomic regulation on circulation and respiration. These are summarized as follows. 1) Orexin-deficient mice exposed to a stressor exhibited an attenuated fight-or-flight response, including increases in respiration and blood pressure and stress-induced analgesia. 2) Stimulation to the amygdala (AMG) or the bed nucleus of the stria terminalis (BNST), both of which are implicated in the stress-induced autonomic responses, induced long-lasting cardiorespiratory excitation in wild-type mice but not in the orexin neuron-ablated mice. Hence, it is likely that the orexin system is one of the essential modulators required for orchestrating the neural circuits controlling autonomic functions and emotional behaviors.
 
Article
Vasostatin I (CgA(1-76)) is a naturally occurring biologically active peptide derived from chromogranin A (CgA), and is so named for its inhibitory effects on vascular tension. CgA mRNA is expressed abundantly in sympathoexcitatory catecholaminergic neurons of the rostral ventrolateral medulla (RVLM). CgA microinjection into the RVLM decreases blood pressure (BP), heart rate (HR) and sympathetic nerve activity (SNA). Proteolytic fragments of CgA are thought to be responsible for the cardiovascular effects observed. We hypothesised that vasostatin I is one of the fragments responsible for the central effects of CgA. We examined the role of a vasostatin I fragment, CgA(17-76) (VS-I((CgA17-76))), containing the portion important for biological effects. The effects of VS-I((CgA17-76)) delivered by intrathecal injection, or microinjection into the RVLM, on cardio-respiratory function in urethane anaesthetised, vagotomised, mechanically ventilated Sprague-Dawley rats (n=21) were evaluated. The effects of intrathecal VS-I((CgA17-76)) on the somato-sympathetic, baroreceptor and peripheral chemoreceptor reflexes were also examined. At the concentrations used (10, 100 or 200 μM, intrathecal; or 5 μM, RVLM microinjection) VS-I((CgA17-76)) produced no change in mean arterial pressure, HR, splanchnic SNA, phrenic nerve amplitude or phrenic nerve frequency. All reflexes examined were unchanged following intrathecal VS-I((CgA17-76)). In the periphery, VS-I((CgA17-76)) potentiated the contractile effects of noradrenaline on rat mesenteric arteries (n=6), with a significant left-shift in the dose response curve to noradrenaline (3.7×10(-7) vs 7.7×10(-7)). Our results indicate that VS-I((CgA17-76)) is active in the periphery but not centrally, and is not a central modulator of cardiorespiratory function and physiological reflexes.
 
Article
Among the many benefits of long-term hormone replacement therapy to postmenopausal women is a significant reduction in risk for and progression of cardiovascular disease. However, long-term estrogen replacement therapy has been associated with several undesirable, and likely dose-dependent, side-effects. There is some evidence to suggest that the dose of estrogen which confers optimal beneficial effects on the cardiovascular system is much lower than that which is currently prescribed for postmenopausal women. The following experiments were conducted to determine the dose-response relationship of acutely administered estrogen on autonomic tone and reflex control of heart rate in ovariectomized Sprague-Dawley female rats. Rats were anaesthetized with sodium thiobutabarbital (100 mg/kg) and instrumented to record blood pressure, heart rate and efferent parasympathetic and sympathetic nerve activities. The sensitivity of the cardiac baroreflex was tested using intravenous injection of either phenylephrine hydrochloride (0.025-0.1 mg/kg) or sodium nitroprusside (0.0025-0.01 mg/kg). Intravenous injection of estrogen produced dose-dependent increases in the magnitude of the baroreflex sensitivity and parasympathetic tone while reducing sympathetic tone with a maximal effect observed at 1 x 10(-3) mg/kg. Prior administration of the selective estrogen receptor antagonist, ICI 182,780 blocked the estrogen-induced changes in baroreflex sensitivity and autonomic tone. These results demonstrate that acutely administered, low-dose estrogen has beneficial effects on autonomic tone and cardiovascular reflexes.
 
Article
Autonomic function was investigated in five affected and five at-risk members of a single kinship of pallidopontonigral degeneration (PPND), which is a progressive syndrome of parkinsonism and frontotemporal dementia resulting from a mutation in the N279K tau gene on chromosome 17. Affected subjects reported symptoms including hyperhidrosis, sialorrhea, urinary frequency or incontinence, thermal intolerance, male sexual dysfunction, lacrimation, and dryness of the eyes or mouth. None had orthostatic hypotension. Autonomic testing revealed mild-to-moderate abnormalities in all five affected subjects and minor abnormalities in the three oldest, asymptomatic, at-risk subjects. Findings in affected subjects consisted of preganglionic sudomotor dysfunction in all five, impaired cardiovagal function in three, and reduced or absent pupillary near responses in four. Tests of adrenergic function were normal in all subjects. The degree of autonomic dysfunction correlated significantly with disease duration and with indices of disease severity. In conclusion, there is evidence in PPND of a disturbance in the central autonomic network.
 
Article
Heptanol and 18 beta-glycyrrhetinic acid (18 beta GA) block gap junctions, but have other actions on transmitter release that have not been characterised. This study investigates the prejunctional and postjunctional effects of these compounds in guinea pig and mouse vas deferens using intracellular electrophysiological recording and confocal Ca(2+) imaging of sympathetic nerve terminals. In mice, heptanol (2 mM) reversibly decreased the amplitude of purinergic excitatory junction potentials (EJPs; 52+/-5%, P<0.05) while having little effect on spontaneous excitatory junction potentials (sEJPs). Heptanol (2 mM) reversibly abolished the nerve terminal Ca(2+) transient in 52% of terminals. 18 beta GA (10 microM) decreased the mean EJP amplitude, and increased input resistance in both mouse (137+/-17%, P<0.05) and guinea pig (354+/-50%, P<0.001) vas deferens indicating gap junction blockade. Further, 18 beta GA increased the sEJP frequency significantly in guinea pigs (by 71+/-25%, P<0.05) and in 5 out of 6 tissues in mice (19+/-3%, P<0.05). Moreover, 18 beta GA depolarised cells from both mice (11+/-1%, P<0.01) and guinea pigs (8+/-1%, P<0.005). Therefore, we conclude that heptanol (2 mM) decreases neurotransmitter release (given the decrease in EJP amplitude) by abolishing the nerve terminal action potential in a proportion of nerve terminals. 18 betaGA (10 microM) effectively blocks the gap junctions, but the increase in sEJP frequency suggests an additional prejunctional effect, which might involve the induction of spontaneous nerve terminal action potentials.
 
Nucleotide sequences of primer sets and annealing temperature (T) were used for RT-PCR reactions. 
The expression of mRNA for IL-19, IL-1β, TNF-α, β-actin and GFAP were carried out using mRNA extracted from LPS-treated and untreated mouse astrocytes cultures and spleen MN cells studied using RT-PCR method. β-actin and GFAP were used as internal control and specific marker of astrocytes, respectively. Experiments were repeated 4 times.  
RT-PCR Reactions for IL-19, IL-1β, TNF-α, GFAP and β-actin were performed using mRNA isolated from cortex of adult C57BL/6 mice following i.p. administration of LPS. β-actin was used as internal control. Experiments were repeated 4 times. The data represent from at least four independent experiments (independently derived astrocyte cultures).  
The purity of astrocyte cultures was determined using indirect immunocytochemical staining with anti-GFAP antibody. (1.A) Confocal Microscopy analysis of primary astroglial cells stained with rabbit anti-GFAP antibody (green). The cell nuclei stained with DAPI (blue). (1.B) Negative control stained with secondary antibody and DAPI. Immunocytochemistry for determination of GFAP, were done for each astrocyte cultures.  
Article
Astrocytes are the most abundant glial cell type. In addition to their neurological roles, astrocytes also have immune functions. They have been involved in antigen presentation in the central nervous system (CNS). Activated astrocytes express adhesion molecules, chemokines and release several inflammatory mediators, pro-inflammatory cytokines, neurotrophic and neuroprotective factors, thus these cells have a dual role within the CNS: neuroinflammation and repair processes. IL-19, IL-20, IL-22, IL-24, IL-26, IL-28A, IL-28B, and IL-29 are members of the IL-10 family of cytokines. These cytokines have different biological functions in spite of partial amino acid sequences homology. Signal transduction of the IL-10 family of cytokines is through R1-type and R2-type receptors. No information has been available about the expression and regulation of IL-19 in mice astrocytes and brain. To investigate the expression of IL-19, we examined its expression in C57BL/6 mice astroglial cells in response to lipopolysaccharide (LPS), using reverse-transcription polymerase chain reaction (RT-PCR) method. We provide for the first time, evidence that astrocytes can express IL-19 mRNA following LPS stimulation. Furthermore, we have found the expression of IL-19 mRNA in the cortex of adult C57BL/6 mice following intraperitoneal (i.p.) administration of LPS. This finding will contribute to current knowledge on the function and behavior of cells and mediators during inflammatory conditions in the brain.
 
Article
In an attempt to know the functional role of alpha1A-adrenoceptors in adipose tissue, white adipocytes (WAT) of Wistar rats were used to investigate the change of glucose uptake after pharmacological activation of alpha1-adrenoceptors. Methoxamine enhanced the uptake of radioactive glucose into isolated WAT in a concentration-dependent manner. Translocation of glucose transporter (GLUT4) from cytosol to membrane was also stimulated with methoxamine. Action of methoxamine to raise glucose uptake was abolished in WAT pre-incubated with the antagonists, both tamsulosin and WB 4101, at concentrations sufficient to block alpha1A-adrenoceptors. However, chlorethylclonidine (CEC). the antagonist of alpha1B-adrenoceptors, showed the inhibition of methoxamine-induced action only at a higher concentration. Even under the treatment with maximal concentration of CEC, methoxamine can produce action about 80% of the vehicle-treated control. The major role of alpha1A-adrenoceptors in the stimulation of glucose uptake by methoxamine can thus be considered. In the presence of specific inhibitor of phospholipase C (PLC), U73312, methoxamine-stimulated glucose uptake into WAT was reduced in a concentration-dependent manner and U73343, the negative control of U73312, did not affect the action of methoxamine. Moreover, chelerythrine and GF 109203X diminished the methoxamine-stimulated glucose uptake at a concentration sufficient to inhibit protein kinase C (PKC). Inhibition of phosphoinositide-3 kinase (PI-3 kinase) by LY294002 also abolished methoxamine-stimulated glucose uptake. Therefore. the obtained data suggest that an activation of alpha1A-adrenoceptors, presence in WAT, by agonist and/or neurotransmitter may increase the glucose uptake via PLC-PKC pathway and the activation of PI-3 kinase.
 
Article
In an attempt to determine whether there is a specific vasopressin receptor (V(1b)) subtype involved in the regulation of body water balance and temperature, vasopressin V(1b) receptor knockout mice were used. Daily drinking behavior and renal excretory function were examined in V(1b)-deficient (V(1b)(-/-)) and control (V(1b)(+/+)) mice under the basal and stress-induced condition. In addition, body temperature and locomotor activity were measured with a biotelemetry system. The baseline daily water intake and urine volume were larger in V(1b)(-/-) mice than in V(1b)(+/+) mice. V(1b)(-/-) mice (V(1b)(-/-)) had significantly higher locomotor activity than wild-type, whereas the body temperature and oxygen consumption were lower in V(1b)(-/-) than in the V(1b)(+/+) mice. Next, the V(1b)(-/-) and V(1b)(+/+) mice were subjected to water deprivation for 48 hr. Under this condition, their body temperature decreased with the time course, which was significantly larger for V(1b)(-/-) than for V(1b)(+/+) mice. Central vasopressin has been reported to elicit drinking behavior and antipyretic action, and the V(1b) receptor has been reported to be located in the kidney. Thus, the findings suggest that the V(1b) receptor may be, at least in part, involved in body water balance and body temperature regulation.
 
Article
Peripheral interleukin-1beta has been implicated in the initiation of fever responses, yet the pathways by which it influences brain function are still unclear. Sectioning the abdominal vagus has been reported to inhibit fever after intraperitoneal administration of interleukin-1beta, suggesting that vagal afferents participate in signaling the brain to mount a fever response to interleukin-1beta. However, the inhibitory effect of subdiaphragmatic vagotomy could be due to alterations in pharmacokinetics such that the intraperitoneally injected cytokine does not reach the general circulation in sufficient quantities to activate the brain via blood-borne signaling. We measured both fever and plasma levels of interleukin-1beta in vagotomized and sham-operated rats after intraperitoneal administration of 1 microg/kg human recombinant interleukin-1beta to determine whether vagotomy reduces fever and levels of circulating interleukin-1beta after intraperitoneal injection. Plasma levels of human recombinant and endogenous rat interleukin-1beta were measured in separate enzyme-linked immunosorbent assays. While intraperitoneal administration of human recombinant interleukin-1beta elevated plasma levels of this cytokine similarly in vagotomized and sham-operated animals, only sham-operated rats responded with fever. Plasma levels of endogenous rat interleukin-1beta were unchanged by any treatment. These results demonstrate that the blockade of intraperitoneal interleukin-1beta-induced fever after subdiaphragmatic vagotomy cannot be accounted for by alterations of interleukin-1beta levels in the general circulation.
 
Renal (A), interscapular brown adipose tissue (IBAT) (B), splenic (C), and lumbar (D) sympathetic nerve discharge (SND) before (Con) and for 60 min after IL-1b administration (100 ng, iv) in rats with intact Fig. 1. Traces of integrated sympathetic nerve discharge (SND) bursts vagus nerves (filled circles) and in cervical-vagotomized rats (open recorded before (Control) and 60 min after IL-1b administration in rats circles). †Significantly different from Con. Z Significantly different from with intact vagus nerves (Intact) and in cervical-vagotomized (Vagotomy) rats with intact vagus nerves (P,0.05). ‡ Significantly different from rats. Horizontal calibration is 500 ms. vagotomized rats. 
Mean arterial pressure (MAP) (left) and heart rate (HR) (right) 
Article
Although the vagus nerve is an important neural pathway mediating immune-to-brain communication, the role of the vagus in mediating sympathetic nerve discharge (SND) responses to peripheral cytokines is not well established. In the present study we determined renal, interscapular brown adipose tissue (IBAT), splenic, and lumbar SND responses before and for 60 min after the intravenous administration of interleukin-1beta (IL-1beta, 100 ng) in chloralose-anesthetized, sham-vagotomized and cervical-vagotomized (bilateral) rats. In sham-vagotomized rats, IL-1beta administration increased (P<0.05) splenic and lumbar SND while renal and IBAT SND remained unchanged from control levels. Renal, splenic, and lumbar SND were increased (P<0.05) whereas IBAT SND remained unchanged from control after IL-1beta in vagotomized rats. Renal, splenic, and lumbar SND responses were significantly higher after IL-1beta in vagotomized compared with sham-vagotomized rats. These results demonstrate that regionally-selective SND (renal, splenic, and lumbar) responses to IL-1beta can occur in the absence of the vagus nerve and suggest that the vagus nerve provides a tonic inhibition to the discharges in these nerves in response to peripheral IL-1beta.
 
Article
Partial elimination of vagal sensory afferents by subdiaphragmatic vagal section has variously been reported to eliminate, to reduce, or to have no effect on fever produced by peripheral lipopolysaccharide and interleukin-1beta (IL-1beta). However, to adequately test the idea that vagal afferents convey immune information to the brain, all vagal input to the central nervous system must be eliminated. This was accomplished by bilateral electrolytic lesions of the nucleus tractus solitarius (NTS). Reflex bradycardia evoked by intravenous phenylbiguanide was eliminated in NTS-lesioned rats, verifying the lesion's effectiveness. IL-1beta (2 microg/kg) was given to conscious, unrestrained rats via an indwelling intraperitoneal catheter and produced rapid fever (approximately 1 degree C) with an onset latency of 15 min and peak response at 30 min, with a second, smaller peak at 130 min. NTS lesions attenuated the first fever peak, with a lesser, non-significant effect on the second peak. The thermogenic capacity of NTS-lesioned rats was evaluated using 3 different strategies: (1) thermogenesis evoked by CNS injections of prostaglandin E2, (2) 3 h exposure to a 4 degrees C environment, and (3) heat production of intrascapular brown fat produced by intravenous infusion of the beta3-adrenergic agonist BRL 37344. NTS-lesioned rats were equivalent, or even superior to control animals in their thermogenic response to these non-immune-related stimuli. Therefore, the impaired febrile response of NTS-lesioned rats to IL-1beta cannot be attributed to reduced thermogenic capacity. Finally, these results suggest that fever elicited by intraperitoneal IL-1beta is, at least in part, dependent on the integrity of NTS neurons, but also that mechanisms independent of vagal afferent projections to the NTS must also play a role in immune-to-brain signaling.
 
Article
Changes in arterial pressure and blood volume are sensed by baroreceptor and vagal afferent nerves innervating aorta and heart with soma in nodose ganglia. The inability to measure membrane potential at the nerve terminals has limited our understanding of mechanosensory transduction. Goals of the present study were to: (1) Characterize membrane potential and action potential responses to mechanical stimulation of isolated nodose sensory neurons in culture; and (2) Determine whether the degenerin/epithelial sodium channel (DEG/ENaC) blocker amiloride selectively blocks mechanically induced depolarization without suppressing membrane excitability. Membrane potential of isolated rat nodose neurons was measured with sharp microelectrodes. Mechanical stimulation with buffer ejected from a micropipette (5, 10, 20 psi) depolarized 6 of 10 nodose neurons (60%) in an intensity-dependent manner. The depolarization evoked action potentials in 4 of the 6 neurons. Amiloride (1 microM) essentially abolished mechanically induced depolarization (15 +/- 4 mV during control vs. 1 +/- 2 mV during amiloride with 20-psi stimulation, n = 6) and action potential discharge. In contrast, amiloride did not inhibit the frequency of action potential discharge in response to depolarizing current injection (n = 6). In summary, mechanical stimulation depolarizes and triggers action potentials in a subpopulation of nodose sensory neurons in culture. The DEG/ENaC blocker amiloride at a concentration of 1 microM inhibits responses to mechanical stimulation without suppressing membrane excitability. The results support the hypothesis that DEG/ENaC subunits are components of mechanosensitive ion channels on vagal afferent and baroreceptor neurons.
 
Article
This brief review will place recent findings on specific neurons and receptors identified as putative central chemoreceptors, namely glutamatergic and serotonergic neurons and purogenic receptors, into the context of our working hypothesis that central chemoreception is a distributed property.
 
Article
Relationship between respiratory morbidity and influenza virus has been well-documented in infants, whatever their age. However, in spite of severe cardiac and respiratory events of central origin, autonomic dysfunction potentially induced by this virus is poorly understood. We thus explored the autonomic nervous system (ANS) profile of infected infants during the 2009 pandemic influenza disease. A group of 10 consecutive infants (5<1 year and 5>1 year) presenting with nvH1N1 infection was evaluated with heart rate variability (HRV) and spontaneous baroreflex (SBR) through a recording during a 15-minute period of quiet sleep as well as 24h holter monitoring. The same recordings were performed in 10 control subjects, paired for gestational and postnatal age. HRV sympathetic and parasympathetic (VLF, LF and HF) indices were obtained using spectral-domain analysis, and spontaneous baroreflex sensitivity (SBR) using simultaneous non-invasive continuous measure of arterial pressure and heart rate. The group of infants less than 1 year had similar value as control group. Conversely, the group of infants more than 1 year showed significant lower spectral total power values (Ptot: 3347 vs. 5926 ms(2)/Hz, p<0.1) and significant lower VLF, LF and HF indices (all p<0.05) than the control group. nvH1N1 infection in child could be associated with severe central autonomic dysfunction. Due to potential severe consequences, a systematic evaluation of autonomic regulation should be performed in order to avoid dramatic events.
 
Article
Both spectral power within the low-frequency component, i.e., 0.04 to 0.15Hz, of systolic pressure and muscle sympathetic nerve activity are increased during head-up tilt. The nerve activity during tilt is altered after space flight and exposure to simulated microgravity. In the present study, correlations of the low-frequency component and the nerve activity were analyzed before and after 20days of -6° of head-down bed rest. Measurements were performed at -6° head-down bed rest, 0° (flat), and 30° and 60° head-up tilt (HUT). Mean arterial pressure during HUT was not different between pre- and post-bed rest, but muscle sympathetic nerve activity in post-bed rest significantly increased at tilt angles of -6°, 0°, 30°, and 60° compared with those during pre-bed rest. The low-frequency component of systolic pressure also significantly increased during post-bed rest compared with pre-bed rest at tilts of 0°, 30°, and 60°. The nerve activity and the frequency component were linearly correlated for individual (r(2)=0.51-0.88) and averaged (r(2)=0.60) values when the values included both pre- and post-bed rest. Thus, the low-frequency component of systolic pressure could be an index of the muscle sympathetic nerve activity during tilt during pre- and post-bed rest.
 
Article
Endothelin-1(1-31) (ET-1(1-31)) is a novel member of the endothelin family, which comprises 31 amino acids and derived from the selective hydrolysis of big ET-1 by chymase. Although ET-1(1-31) has been reported to be involved in biological effects via direct or indirect (converting to ET-1(1-21)) mechanisms, the cardiovascular effects of central ET-1(1-31) are not fully identified. The present study was designed to comparatively investigate the cardiovascular effects of intracerebroventricular (icv) application of ET-1(1-31) or ET-1(1-21) in anesthetized rats. Injection (icv) of ET-1(1-31) (500 pmol) produced a biphasic blood pressure response: an initial increase (from 118+/-8 to 138+/-14 mmHg, P<0.05) followed by a sustained decrease in BP (from 118+/-8 to 58+/-9 mmHg, P<0.05), which was very similar to BP response to icv injection of big ET-1 (500 pmol) or ET-1(1-21) (25 pmol)(.) The cardiovascular effects of icv injection of ET-1(1-31) or ET-1(1-21) were completely antagonized by ET(A) receptor antagonist BQ123 but not ET(B) receptor antagonist BQ788. Furthermore, pretreatment with ET converting enzyme inhibitor phosphoramidon (10 nmol) abolished the cardiovascular effects evoked by icv injection of ET-1(1-31) or big ET-1. In conclusion, the current data showed that central ET-1(1-31) produced the similar cardiovascular effects as those of central ET-1(1-21), and suggesting that the central cardiovascular effects of ET-1(1-31) resulted from it converting to ET-1(1-21) and then activating ET(A) receptors.
 
Article
Viktor Hamburger has just died at the age of 100. It is 50 years since he and Rita Levi-Montalcini laid the foundations for the study of naturally occurring cell death and of neurotrophic factors in the nervous system. In a period of less than 10 years, from 1949 to 1958, Hamburger and Levi-Montalcini made the following seminal discoveries: that neuron cell death occurs in dorsal root ganglia, sympathetic ganglia and the cervical column of motoneurons; that the predictions arising from this observation, namely that survival is dependent on the supply of a trophic factor, could be substantiated by studying the effects of a sarcoma on the proliferation of ganglionic processes both in vivo and in vitro; and that the proliferation of these processes could be used as an assay system to isolate the factor. This work provides a short review mostly of the early history of this subject in the context of the Hamburger/Levi-Montalcini paradigm. This acts as an introduction to a consideration of models that have been proposed to account for how the different sources of growth factors provide for the survival of neurons during development. It is suggested that what has been called the 'social-control' model provides the most parsimonious quantitative description of the contribution of trophic factors to neuronal survival, a concept for which we are in debt to Viktor Hamburger and Rita Levi-Montalcini.
 
Article
The present study was aimed at investigating whether the blood pressure-R-R interval relation obtained by ABPM may give useful information about autonomic control in the 24 h period. To this purpose ABPM was performed in 60 healthy young subjects (30 females and 30 males, mean age 21.8+/-1.0 years) and the collected variables were copied to a software program to convert heart rate into R-R interval values, for statistical analysis and graphic representation. The following data were calculated: 1) day and night means+/-SD; 2) difference and percent difference in mean night less mean day R-R interval (Delta y), diastolic and systolic blood pressures (Delta x) and their Delta y/Delta x ratios; 3) intercept (a_24 h), slope (b_24 h) and r coefficient (r_24 h) of the linear regressions of 24 h R-R interval over diastolic and systolic blood pressure values. In all subjects night, with respect to day, was characterized by R-R interval lengthening and blood pressure lowering. Despite this common pattern, day and night means and SDs, night and day differences, Delta y/Delta x ratios, a_24 h and b_24 h were different from individual to individual, but they were characteristic and reproducible in 20 out of the 21 subjects in which ABPM was repeated twice. Subjects could thus be classified according to their Delta y/Delta x ratios and slope (b_24 h). The 24 h blood pressure-R-R interval relation as calculated from ABPM yields individually characteristic indices of circadian sympatho-vagal reciprocity. This novel approach may be helpful in characterizing the 24 h autonomic control of several groups of patients.
 
Article
It has recently been demonstrated that SDNN of heart rate variability (HRV) is a useful independent prognostic tool in chronic heart failure (CHF). The purpose of the present study was to evaluate if spectral and non-linear analysis of 24-h HRV, considered markers of autonomic cardiac modulation, contain independent prognostic information in CHF patients. Twenty normal subjects and thirty consecutive outpatients with clinically stable CHF were studied for 2 years. Periods of 300 R-R intervals were analyzed from Holter recordings. The power spectral analysis, the slope of the linear relationship between log-power versus log-frequency (1/f), and the complexity content (using corrected conditional entropy; CCE) of the R-R series were calculated. The normalized power of the low frequency spectral component (LF) and the 1/f slope were significantly lower in patients compared to controls (respectively 30.1 +/- 3.0 vs. 48.6 +/- 3.4 and -1.27 +/- 0.04 vs. -1.08 +/- 0.05; P < 0.05). Moreover, the patients who died during the study presented a reduced LF (20.9 +/- 4.1 vs. 35.5 +/- 3.5 nu; P < 0.05) and a steeper 1/f slope (-1.40 +/- 0.09 vs. -1.21 +/- 0.04 nuts, P < 0.05) compared to survivors. These results remained significant in a logistic model including heart rate and SDNN. The information content present in spectral and non-linear analysis of HRV in CHF patients has prognostic relevance independently from the time domain measures of HRV. In particular, the reduction of LF power seems the best indicator among those considered.
 
Article
The facial feedback hypothesis suggests that feedback from cutaneous and muscular afferents influences our emotions during the control of facial expressions. Enhancing facial expressiveness produces an increase in autonomic arousal and self-reported emotional experience, whereas limiting facial expression attenuates these responses. The present study investigated differences in autonomic responses during imitated versus observed facial expressions. Thus, we obtained the facial electromyogram (EMG) of the corrugator muscle, and measured the skin conductance response (SCR) and pupil size (PS) of participants while they were either imitating or simply observing emotional expressions of anger. We found that participants produced significantly greater responses across all three measures (EMG, SCR, and PS) during active imitation than during passive observation. These results show that amplified feedback from facial muscles during imitation strengthens sympathetic activation in response to negative emotional cues. Our findings suggest that manipulations of muscular feedback could be used to modulate the bodily expression of emotion, including autonomic responses to the emotional cues.
 
Article
In this experiment, we examined the influence of the posterior hypothalamic adenosine A(2A) receptors on the central cardiovascular regulation of blood pressure (BP) and heart rate (HR). Posterior hypothalamic injection of drugs was performed in anesthetized, artificially ventilated male Sprague-Dawley rats. Injection of CGS-21680HCl (CGS; 20 nmol), an adenosine A(2A) receptor agonist, elicited a decrease of arterial BP and HR, while injection of 8-(3-Chlorostyryl)caffeine (CSC; 10 nmol), an adenosine A(2A) receptor antagonist, blocked the depressor and bradycardiac effects of CGS (20 nmol). To examine the mechanisms of cardiovascular regulation of adenosine A(2A) receptors in the posterior hypothalamus, we applied the adenylate cyclase and guanylate cyclase inhibitors, to the posterior hypothalamus. Pretreatment with MDL-12,330 (MDL; 10 nmol), an adenylate cylase inhibitor, attenuated the depressor and bradycardiac effects of CGS. However, pretreatment with, LY-83,583 (LY; 5 nmol), a soluble guanylate cyclase inhibitor, did not alter the effects of CGS. Additionally, we examined the modification of the cardiovascular effects of adenosine A(2A) receptors through the ATP-sensitive K+ channel in the posterior hypothalamus. Posterior hypothalamic administration of glipizide (20 nmol) significantly attenuated the cardiovascular depressor actions elicited by CGS. These results suggest that adenosine A(2A) receptors in the posterior hypothalamus play an inhibitory role in central cardiovascular regulation, and that adenylate cyclase, but not guanylate cyclase, mediates the depressor and bradycardiac actions of adenosine A(2A) receptors. Furthermore, ATP-sensitive K+ channels mediate the posterior hypothalamic cardiovascular regulation of adenosine A(2A) receptors.
 
Article
Many diseases are related to cerebrospinal fluid (CSF) hydrodynamics. Therefore, understanding the hydrodynamics of CSF flow and intracranial pressure is helpful for obtaining deeper knowledge of pathological processes and providing better treatments. Furthermore, engineering a reliable computational method is promising approach for fabricating in vitro models which is essential for inventing generic medicines. A Fluid-Solid Interaction (FSI)model was constructed to simulate CSF flow. An important problem in modeling the CSF flow is the diastolic back flow. In this article, using both rigid and flexible conditions for ventricular system allowed us to evaluate the effect of surrounding brain tissue. Our model assumed an elastic wall for the ventricles and a pulsatile CSF input as its boundary conditions. A comparison of the results and the experimental data was done. The flexible model gave better results because it could reproduce the diastolic back flow mentioned in clinical research studies. The previous rigid models have ignored the brain parenchyma interaction with CSF and so had not reported the back flow during the diastolic time. In this computational fluid dynamic (CFD) analysis, the CSF pressure and flow velocity in different areas were concordant with the experimental data.
 
Photographs of coronal sections throughout the CA1 area of hippocampus in all groups (Cresyl violet staining ×40, neurons are blue) A: Control-saline B: MDMA 2.5 mg/kg C: MDMA 5 mg/kg D: MDMA 10 mg/kg The highest decrease in the number of neurons was shown in response to MDMA with the dose of 5 mg/kg compared to the control-saline 
Photographs of coronal sections throughout the CA1 area of hippocampus in all groups (PTAH staining ×40, Astrocytes are purple) A: Control-saline B: MDMA 2.5 mg/kg C: MDMA 5 mg/kg D: MDMA 10 mg/kg 
Significant difference in the number of neurons between sham (Control-saline) and MDMA treated groups in all areas of the hippocampus 
Significant difference in the number of astrocytes between sham (Control-saline) and MDMA treated groups in all areas of the hippocampus 
Article
Introduction MDMA or ecstasy is a derivative of amphetamines used mostly by young people worldwide. Although the acute effects of this drug are known, the effect of chronic administration is not well studied. Therefor the aim of this study was to determine the effects of repeated (long term) administration of MDMA on rats’ memory and their hippocampal cell density. Method Young adult male Wistar rats 200 ± 20 g served as subjects. The rats were randomly distributed into three MDMA treated groups (3×2.5 mg/kg, 3×5 mg/kg, 3×10 mg/kg) and one control-saline group. All animals received MDMA intraperitoneally (3h apart; a challenge) 7th day of every week for consecutive 4 weeks. Animals were trained before and were tested after injections for their memory status using the standards passive avoidance method. Finally, 24hr after the memory test, rats were sacrificed and after tissue operations, the hippocampal astrocytes and neurons were counted. Results Results showed that the number of neurons in all experimental groups was lower than the control-saline group. The most decreased number of neurons was shown in 5 mg/kg MDMA group compared to control-saline in all the regions of hippocampus. Also we found that repeated administration of MDMA reduced the number of hippocampal astrocytes. Discussion It is concluded that repeated administration of MDMA can reduce density of neurons and astrocytes and this decrease is not dose dependence.
 
Article
It is well documented that the intrinsic enteric nervous system of the gastrointestinal (GI) tract sustains neuronal losses and reorganizes as it ages. In contrast, age-related remodeling of the extrinsic sympathetic projections to the wall of the gut is poorly characterized. The present experiment, therefore, surveyed the sympathetic projections to the aged small intestine for axonopathies. Furthermore, the experiment evaluated the specific prediction that catecholaminergic inputs undergo hyperplastic changes. Jejunal tissue was collected from 3-, 8-, 16-, and 24-month-old male Fischer 344 rats, prepared as whole mounts consisting of the muscularis, and processed immunohistochemically for tyrosine hydroxylase, the enzymatic marker for norepinephrine, and either the protein CD163 or the protein MHCII, both phenotypical markers for macrophages. Four distinctive sympathetic axonopathy profiles occurred in the small intestine of the aged rat: (1) swollen and dystrophic terminals, (2) tangled axons, (3) discrete hyperinnervated loci in the smooth muscle wall, including at the bases of Peyer's patches, and (4) ectopic hyperplastic or hyperinnervating axons in the serosa/subserosal layers. In many cases, the axonopathies occurred at localized and limited foci, involving only a few axon terminals, in a pattern consistent with incidences of focal ischemic, vascular, or traumatic insult. The present observations underscore the complexity of the processes of aging on the neural circuitry of the gut, with age-related GI functional impairments likely reflecting a constellation of adjustments that range from selective neuronal losses, through accumulation of cellular debris, to hyperplasias and hyperinnervation of sympathetic inputs.
 
Article
Current evidence indicates that rises in systemic levels of estrogen create in the uterus an inhibitory environment for sympathetic nerves. However, molecular insights of these changes are far from complete. We evaluated if semaphorin 3F mRNA, a sympathetic nerve repellent, was produced by the rat uterus and if its expression was modulated by estrogen. We also analyzed whether uterine nerves express the semaphorin 3F binding receptor, neuropilin-2. Uterine levels of semaphorin 3F mRNA were measured using real time reverse transcriptase-polymerase chain reaction in prepubertal rat controls and following chronic estrogen treatment. Localization of semaphorin 3F transcripts was determined by in situ hybridization and the expression of neuropilin-2 was assessed by immunohistochemistry. These studies showed that: (1) chronic estrogen treatment led to a 5-fold induction of semaphorin 3F mRNA in the immature uterus; (2) estrogen provoked a tissue-specific induction of semaphorin 3F which was particularly localized in the connective tissue that borders muscle bundles and surrounds intrauterine blood vessels; (3) two major cell-types were recognized in the areas where transcripts were concentrated, fibroblast-like cells and infiltrating eosinophil leukocytes; and (4) some delicate nerve terminal profiles present in the estrogenized uterus were immunoreactive for neuropilin-2. Temporal and spatial expression patterns of semaphorin 3F/neuropilin-2 are consistent with a possible role of this guidance cue in the remodeling of uterine sympathetic innervation by estrogen. Though correlative in its nature, these data support a model whereby semaphorin 3F, in combination with other inhibitory molecules, converts the estrogenized myometrium to an inhospitable environment for sympathetic nerves.
 
Relative expression of Epac in 1321N1 (Fig. 1A) and U87MG (Fig. 1B) cell lines in un-treated (C) and treated cells with 10 μg/ml fluoxetine (F) HPRT1 was used as internal control.  
Fluoxetine effects on Cx43 mRNA expression with and without cAMP-PKA signaling pathway. 1321N1 cells treated with F (Fluoxetine), SF (AC inhibitor+ Fluoxetine) and F+H89 (Fluoxetine + PKA inhibitor) for 24 hours and changes in transcript amount expression were determined by real-time qPCR. It revealed an up-regulation of Cx43 in all of groups (*p < 0.01 compared to untreated controls).  
Expression of Cx43 protein in the 1321N1 cells. Cx43 protein levels were normalized to that of β-actin protein . Data are expressed as means ± S.E.M. Each immunoblotting was performed in duplicate to increase the reliability of the measurements. Lower Panels show representative immunoblots of Cx43 (43 kDa). C (un-treated), F (  
Article
Recent studies have shown that astrocytes play major roles in normal and disease condition of the central nervous system including multiple sclerosis (MS). Molecular target therapy studies in MS have revealed that connexin-43 (Cx43) and Aquaporin-4 (AQP4) contents of astrocytes undergo expression alteration. Fluoxetine had some effects in MS patients unrelated to its known antidepressant effects. Some of fluoxetine effects were attributed to its capability of cAMP signaling pathway stimulation. This study aimed to investigate possible acute effects of fluoxetine on Cx43 and AQP4 expression in astrocyte. Astrocytoma cells were treated for 24 hours with fluoxetine (10 and 20 µg/ml) with or without adenyl cyclase (AC) and protein kinase A (PKA) inhibition. Cx43 expression at both mRNA and protein levels and AQP4 expression at mRNA level were evaluated. Acquired results showed that fluoxetine with and without AC and PKA inhibition resulted in Cx43 up-regulation both in mRNA and protein levels, whereas AQP4 expression have not changed. In conclusion, data showed that fluoxetine alone and in the absence of serotonin acutely up-regulated Cx43 expression in astrocytes that can be assumed in molecular target therapy of MS patients. It seems that cAMP involvement in fluoxetine effects need more researches.
 
Article
Although extracorporeal shockwave has been applied in the treatment of various diseases, the biological basis for its analgesic effect remains unclear. Therefore, we investigated the dorsal root ganglion neurons of rats following shockwave exposure to the footpad to elucidate its effect on the peripheral nervous system. We used activating transcription factor 3 (ATF3) and growth-associated phosphoprotein (GAP-43) as markers for nerve injury and axonal regeneration, respectively. The average number of neurons immunoreactive for ATF3 increased significantly in the treated rats at all experimental time points, with 78.3% of those neurons also exhibiting immunoreactivity for GAP-43. Shockwave exposure induced injury of the sensory nerve fibers within the exposed area. This phenomenon may be linked to the desensitization of the exposure area, not the cause of pain, considering clinical research with a particular absence of painful adverse effect. Subsequent active axonal regeneration may account for the reinnervation of exposed area and the amelioration of the desensitization.
 
Article
Cancer chemotherapy drugs, such as cisplatin, are extremely potent for producing nausea and vomiting. The acute effects of these treatments are partly controlled using anti-emetic drugs, but the delayed effects (>24 h), especially nausea, are much more difficult to treat. Furthermore, cisplatin induces a long-term (up to 48 h) increase in pica in rats. Pica is manifested as an increase in consumption of kaolin (clay) and is used as a measure of visceral sickness. It is unknown what brain pathways might be responsible for this sickness associated behavior. As a first attempt to define this neural system, rats were injected (i.p.) with 3, 6, or 10 mg/kg cisplatin (doses reported to produce pica) and sacrificed at 6, 24, or 48 h to determine brain Fos expression. The primary results indicate: 1) increasing the dose of cisplatin increased the magnitude and duration of brain Fos expression, 2) most excitatory effects on hindbrain nucleus of the solitary tract (NTS) and area postrema (AP) Fos expression occurred within 24 h after cisplatin injection, 3) 6 and 10 mg/kg cisplatin treatment produced large increases in Fos expression in the central amygdala (CeA) and bed nucleus of the stria terminalis (BNST), including 48 h after injection, and 4) cisplatin treatment produced little effect on Fos expression in the paraventricular and supraoptic nuclei of the hypothalamus. These results indicate that cisplatin activates a neural system that includes the dorsal vagal complex (NTS and AP), CeA, and BNST.
 
Article
Neuroendocrine secretory protein 55 (NESP55) is a soluble, acidic and heat-stable protein, belonging to the class of chromogranins. It is expressed specifically in endocrine cells and the nervous system, and is probably involved in both constitutive and regulated secretion. In the present study, we investigated the distribution of NESP55 in various rat sympathetic ganglia by immunohistochemistry. The expression of NESP55-IR was detected in a subpopulation of principal neurons in the rat SCG, which was also TH positive, and, thus, adrenergic. In the rat stellate ganglion, more than two thirds of NESP55 positive neurons were adrenergic. Colocalization of NESP55 and calcitonin gene-related peptide (CGRP) in cholinergic neurons was also observed. In the rat thoracic chain, however, the majority of NESP55 positive neurons appeared to lack TH. No detectable NESP55-IR was found in the mouse SCG. Furthermore, in the sexually dimorphic SCG, it was demonstrated that, 80% of the NESP55 positive principal neurons were also NPY positive in the male rat, while a slightly higher, but statistically significant proportion, 87%, was found in the female. Whether or not this small difference is physiologically significant is unknown. The present data provide basic knowledge about the expression of NESP55 in the sympathetic autonomic nervous system of rat, which may further our understanding of the functional significance of NESP55.
 
Article
To determine the effect of aging on the cardiovascular response to postural change, we examined the cardiovascular sympathetic and parasympathetic response to active standing in 610 healthy Japanese subject (6-83 years) measuring the initial heart rate (HR) response for 3 min in the supine and standing position, we also measured the coefficient of variation of R-R interval (CV(R-R)). As a result, the cardiovascular response to active standing demonstrated a different change with aging between sympathetic and parasympathetic. Sympathetic function was in a sthenia state in young subjects, and that this function declined with age increasing. Whereas, parasympathetic function was immature enough to inhibit the sympathetic tone in young subjects and matured at 20 years of age, and had an ability to inhibit sympathetic tone. CV(R-R) show a linear change that decline with age increasing. These results indicated that the cardiovascular parasympathetic response to active standing shows a characteristic change with aging that differs from cardiovascular parasympathetic at rest represented by CV(R-R). The present study is the first report to demonstrate the cardiovascular response to standing in relation to aging in large population. These results suggested that the cardiovascular response to postural change is dependent on subject's age.
 
Article
The effect of hepatic branch vagotomy on liver regeneration induced by ursodesoxycholic acid was examined in 66% hepatectomized rats. A standard feeding was obtained from animals with hepatic branch vagotomy, and all rats examined received pair food intake. Liver regeneration was evaluated by the hepatocyte mitotic index. When ursodesoxycholic acid 12.5 mg kg(-1) day(-1) was administered orally, a significant increase in the mitotic index was observed 2 and 3 days after hepatectomy, and the mitotic index response 2 days after hepatectomy was dose-dependent in the range of 0-25 mg kg(-1) day(-1). Hepatic branch vagotomy suppressed the mitotic index 2 days after hepatectomy, and the mitotic index increase due to ursodesoxycholic acid was blocked by hepatic branch vagotomy, but not by gastric branch vagotomy. It was also noted that hepatic branch vagotomy or ursodesoxycholic acid or both did not affect serum parameters indicating liver and kidney function. Because ursodesoxycholic acid exists in the bile juice, these results suggest that the hepatic vagus nerve is specifically active with endogenous bile acids in the control of liver regeneration.
 
Article
We have studied the influence of temperature and ARL 67156 on ATP hydrolysis in mouse and guinea pig vas deferens in order to explore the properties of the enzymatic inactivation mechanism proposed to regulate purinergic neurotransmission at the sympathetic neuromuscular junction of smooth muscle. The ectonucleotidase activity was determined by using the malachite green method to measure the inorganic phosphate (Pi) liberated with ATP used as a substrate. ATP hydrolysis in both species was found to be insensitive to ouabain (100 microM), sodium azide (1 mM), sodium vanadate (100 microM) and beta-glycerophosphate (10 mM) and was also found to depend on Ca2+ and Mg2+. V(MAX) of the ectonucleotidase activity for guinea pig and mouse vas deferens was 958.4+/-66.3 and 79.7+/-8.5 pmol/min/mg, while K(M) was 625.1+/-45.2 and 406.0+/-29.0 microM, respectively. Cooling the tissues from 35 to 25 degrees C reduced the enzyme activity significantly (P<0.01) by 52.7+/-9.2% in guinea pig vas deferens and 34.9+/-5.3% in mouse vas deferens. ARL 67156 (100 microM), the specific ecto-ATPase inhibitor, caused a reduction in enzyme activity in guinea pig and mouse vas of 54.1+/-16.4% and 53.0+/-7.6%, respectively (P<0.01). The degree of inhibition of ATP hydrolysis by lowered temperature and 100 microM ARL 67156 correlates well with the reported potentiation and prolongation of junction potentials under these conditions. It is concluded that ecto-ATPase or a closely related ectonucleotidase plays an important role in the physiological regulation of purinergic neurotransmission.
 
Article
Urodynamic and pharmacological studies were performed to investigate the effect of crystalluria on the micturition reflex and the involvement of glutamatergic transmission. The rats, which were given LP-805 (100 mg/kg/day) orally for 12 days, voided crystalluria. The pH of these crystalluria (LP-805 urine) was the same as normal urine. The amount of crystals was 70-100/division magnified 400 x. The end of the crystals was sharp. Intravesical administration of LP-805 urine induced hyperreflexia of the micturition reflex in normal rats. When the infusion solution was changed to LP-805 urine from saline, the latency was reduced to 57.6+/-2.1% of control in single cystometrogram (CMG) or was reduced to 51.4+/-0.9% of control in continuous CMG. The voiding volume was reduced to 52.1+/-3.6% of control in single CMG or was reduced to 62.5+/-0.8% of control in continuous CMG. These parameters were recovered after LP-805 urine was removed. Intravesical administration of acetic acid did not induce hyperreflexia of the micturition reflex in LP-805-treated rats. These data suggest that the chronic irritation by aculeate crystals might induce hyperreflexia of the micturition reflex, which increase afferent neuronal activity. Intravenous administration of MK-801 (0.001 to 1 mg/kg) inhibited the micturition reflex in a dose-dependent manner. The ID50 in LP-805-treated rats (0.03 mg/kg i.v.) was lower than that in normal rats (0.56 mg/kg i.v.). After chronic irritation of the bladder epithelium, MK-801 sensitivity was enhanced for the micturition reflex. These data suggested that crystalluria elicit hyperreflexia in the micturition reflex that mediated with NMDA glutamatergic receptors.
 
Article
The positive outcome that hypothermia contributes to brain and cardiac protection following ischemia has stimulated research in the development of pharmacological approaches to induce a hypothermic/hypometabolic state. Here we review three papers to highlight the role of the adenosine 1 receptor (A1AR) as a potential mediator and physiological regulator of a hypothermic state in both hibernating and non-hibernating mammals. We would like to emphasize the importance of comparative studies between hibernating and non-hibernating species that could lead to important discoveries on the mechanisms inducing hibernation and how they might be translated to induce a clinically useful hypothermic state.
 
Article
Several findings suggest that A1 noradrenergic neurons in the caudal ventrolateral medulla (CVLM) contribute to body fluid homeostasis and cardiovascular regulation. Recently we demonstrated that the renal vasodilation induced by infusion of hypertonic saline (HS) depends on the integrity of the A1 neurons. Here we determined the effect of lesions of these neurons on the inhibition of the renal sympathetic nerve activity (RSNA) induced by HS infusion. All experiments were performed in Wistar rats (280-350 g). A1 neurons were lesioned by microinjections of antidopamine-beta-hydroxylase-saporin (6.3 ng in 60 nl) into the CVLM (n=5), whereas sham rats received microinjections of free saporin (1.3 ng in 60 nl, n=10). Two weeks later, rats were anesthetized (urethane 1.2 g/kg, iv), and instrumented for recording of arterial pressure and RSNA. In sham rats, HS infusion (3 M NaCl, 0.18 ml/100 g bw, iv) induced a transient (</=30 min) hypertension (peak at 10 min; 9+/-5 mm Hg) and a fall in RSNA (-32+/-7% of baseline at 10 min). A1-lesions increased the duration of the pressor response induced by HS infusion (16+/-2 mm Hg at 60 min) and abolished the fall in RSNA (-6+/-8% of baseline at 10 min). Catecholaminergic lesions extensions were confirmed by immunocytochemistry. Unilateral renal denervation reduced the renal vasodilatation induced by HS infusion (112+/-7% in denervated rats versus 127+/-4% in sham, 20 min after HS). These results suggest that A1 noradrenergic neurons are involved in the sympathoinhibition and consequent renal vasodilatation to acute changes in the extracellular fluid compartment.
 
Top-cited authors
Winfried Neuhuber
  • Friedrich-Alexander-University of Erlangen-Nürnberg
Ronald P Gaykema
  • University of Virginia
Lisa E Goehler
  • University of Virginia
Linda Watkins
  • University of Colorado Boulder
Robert D Foreman
  • University of Oklahoma Health Sciences Center