Atrial natriuretic peptide in the subfornical organ reduces drinking induced by angiotensin or in response to water deprivation.

Three experiments tested whether the subfornical organ (SFO) could be a site of action for the antidipsogenic effects of atrial natriuretic peptide (ANP) in rats. Pretreatment with 100, 230, or 500 pmol ANP in the SFO reduced drinking induced by 10 pmol angiotensin II in the SFO. Drinking in response to water deprivation was reduced by ANP in rats having cannulas in or near the SFO, but not in rats having cannulas distant from the SFO or in the ventricles. Finally, ANP had no effect on eating or drinking after food deprivation, suggesting that the rats in the other experiments were not acutely incapacitated. The SFO may mediate the central effects of ANP on drinking induced by angiotensin or in response to water deprivation and could play a similar role in the central effects of ANP on salt appetite, diuresis, vasopressin secretion, and blood pressure. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Regional differences in the morphology of the rat subfornical organ

Properties of whole milk and milk fractions from cows fed a diet that gave a greatly increased proportion of unsaturated fatty acid residues (especially of linoleic acid) in the milk lipids were studied, and this milk (high-linoleic milk) was compared with milk from cows on a control diet (control milk). The milk fractions were isolated by high-speed centrifugation of whole milk or cream and were examined by chemical analysis and electron microscopy. During centrifugation the globules of milk fat were disrupted and the membranes (fat-globule 'ghosts') floated as a layer beneath the free lipid. Membrane proteins from the 2 sorts of milk gave the same electrophoretic pattern and the amino acid compositions were the same. Lipid analysis of the membrane fraction from high-linoleic milk showed the expected increase in the proportion of unsaturated fatty acid residues in the neutral lipids, but there was an unexpected decrease in the proportion of unsaturated residues in the membrane phospholipids. No differences were found between high-linoleic and control milk in the ultrastructure of the milk-fat globules or the isolated membranes.     

Effect of glutamate and angiotensin II on whole cell currents and release of nitric oxide in the rat subfornical organ

Blood-borne angiotensin II (AngII) is known to mediate water-intake by its excitatory effect on neurons in the subfornical organ (SFO). Conversely, nitric oxide (NO) has exclusively inhibitory effects on rat SFO-neurons and on SFO-mediated water-intake. Extracellular and patch-clamp recordings from freshly dissociated rat SFO-neurons showed that glutamate activates AngII-sensitive SFO-neurons by opening ligand-gated cation channels. An immunocytochemical study showed that activation of glutamate receptors increased the concentration of the inhibitory second messenger cGMP in the SFO. A model is proposed suggesting that NO protects SFO-neurons from overexcitability by excitatory neurotransmitters.     

Effects of subfornical organ extracts of salt-water balance in the rat

The subfornical organ (SFO) is regarded as a neurosecretory structure but no information is available on the nature or biological effects of the secretory products(s). Supernatants of water homogenates of rat SFO were lyophilized and reconsittuted in artificial cerebrospinal fluid (CSF). Intracerebroventricular (IVT), but not subcutaneous, administration of this material to rats produced diuresis, natriuresis and kaliuresis in the following 8 h daylight period. During the overnight cycle, consummatory behavior and excretion of sodium and potassium were reduced. Similar responses were obtained after IVT administration of cerebellar cortex (CB) or large amounts of plasma. SFO, CB and cerebral cortex (CC) were incubated in potassium-enriched CSF to enhance release of secretory products. Urine volume was increased 8 h after IVT injection of SFO media; in the overnight cycle, food consumption, absolute urinary sodium and potassium, and [Na+-a1 were reduced. These effects were not produced by IVT injection of CC or CB media, or equal amounts of plasma proteins. Additional experiments demonstrated that choroid plexi and SFO effects were similar and that the active SFO material was dialyzable and thermal stable. These data suggest that SFO contains a water-soluble substance which is released into a posassium-enriched medium. The material is heat stable, has a relatively low molecular weight, and alters salt-water balance after injection into ventricular cerebrospinal fluid.     

Localization of receptors for the dipsogenic action of angiotensin II in the subfornical organ of rats.

Investigated the proposal that the subfornical organ (SFO) is a site of receptors for drinking induced by angiotensin II (AII). A total of 159 male albino Holtzman rats was used. Intracranial injections of physiological doses of AII elicited drinking if and only if applied directly to the SFO (Exp I). Ablation of the SFO selectively (Exp II) and permanently (Exp IV) eliminated drinking elicited by physiological doses of iv infused AII. Animals in which SFO had been ablated responded normally to cellular dehydration but reduced responding to the extracellular thirsts of beta-adrenergic activation and hyperoncotic colloid dialysis (Exp III). Infusion of saralasin, an AII antagonist, directly into the SFO selectively and reversibly antagonized iv AII drinking (Exp V). The hypothesis that the SFO contains dipsogenic receptors for circulating AII was supported. (2 p ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Differential neuronal responses to angiotensin III from the subfornical organ of normotensive and spontaneously hypertensive rats

We previously reported that chronic central administration of angiotensin III (AIII) fails to produce sustained drinking behavior in spontaneously hypertensive rats (SHR), possibly because of the development of early desensitization of the angiotensin receptors. The present study extended these findings to the cellular level, using brain-slice preparation from Wistar-Kyoto rats (WKY) and SHR, in conjunction with single-neuron recording in the subfornical organ (SFO), a target site for angiotensin II-induced drinking. We found that a majority of the SFO neurons studied (13/18 in WKY, 20/28 in SHR) responded in a dose-related manner to AIII, given in the range of 10(-6)-10(-5) M. This excitation was receptor-specific, since it was reversed by Ile7-AIII (10(-4)-10(-3) M), the selective AIII antagonist. Bestatin (10(-5)-10(-4) M), an aminopeptidase B inhibitor, did not discernibly affect basal spike frequency when delivered alone. Nevertheless, given in combination with the heptapeptide, bestatin reduced the intensity and duration of SFO neuronal response in WKY to the higher dose (10(-5) M), and in SHR to both doses (10(-6) or 10(-5) M), of AIII. These data suggest that the SFO may also be a central site of action for AIII. Moreover, prolonging the action of AIII by protecting it from being metabolized with bestatin may produce desensitization of the angiotensin receptors on SFO neurons. This was particularly so in the SHR, which are thought to be defective in the degradation of the heptapeptide in the brain.     

Interaction of hydration and subfornical organ lesions in sodium-depletion induced salt appetite.

The authors tested whether the level of hydration after furosemide diuresis and 22 hrs of sodium depletion affects the amount of water or 0.3 M NaCl solution consumed by rats with intact brains or with lesions of the subfornical organ (SFO). Rats received 2 (underhydrated) or 10 (euhydrated) ml/kg water by gavage as the only fluid input 2, 4, and 20 hrs after 10 mg/kg furosemide. These hydration treatments had little or no effect on the amount of saline consumed in 2 hrs by intact rats. SFO lesions reduced water intake regardless of hydration condition. Euhydrated, SFO-lesioned rats drank a normal amount of saline, but underhydrated, lesioned rats drank less saline than any other group. Thus, euhydration may facilitate salt appetite in SFO-lesioned rats, and the deficits in salt appetite noted in SFO-lesioned rats may result from deficits in water ingestion rather than from a destruction of angiotensin II receptor sites that directly provoke salt appetite. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Role of angiotensin II and vasopressin receptors within the supraoptic nucleus in water and sodium intake induced by the injection of angiotensin II into the medial septal area

In this study we investigated the effects of the injection into the supraoptic nucleus (SON) of non-peptide AT1- and AT2-angiotensin II (ANG II) receptor antagonists, DuP753 and PD123319, as well as of the arginine-vasopressin (AVP) receptor antagonist d(CH2)5-Tyr(Me)-AVP, on water and 3% NaCl intake induced by the injection of ANG II into the medial septal area (MSA). The effects on water or 3% NaCl intake were assessed in 30-h water-deprived or in 20-h water-deprived furosemide-treated adult male rats, respectively. The drugs were injected in 0.5 microliter over 30-60 s. Controls were injected with a similar volume of 0.15 M NaCl. Antagonists were injected at doses of 20, 80 and 180 nmol. Water and sodium intake was measured over a 2-h period. Previous administration of the AT1 receptor antagonist DuP753 into the SON decreased water (65%, N = 10, P < 0.01) and sodium intake (81%, N = 8, P < 0.01) induced by the injection of ANG II (10 nmol) into the MSA. Neither of these responses was significantly changed by injection of the AT2-receptor antagonist PD123319 into the SON. On the other hand, while there was a decrease in water intake (45%, N = 9, P < 0.01), ANG II-induced sodium intake was significantly increased (70%, N = 8, P < 0.01) following injection of the V1-type vasopressin antagonist d(CH2)5-Tyr(Me)-AVP into the SON. These results suggest that both AT1 and V1 receptors within the SON may be involved in water and sodium intake induced by the activation of ANG II receptors within the MSA. Furthermore, they do not support the involvement of MSA AT2 receptors in the mediation of these responses.     

Differential regulation of angiotensinogen and AT1A receptor mRNA within the rat subfornical organ during dehydration

The present study describes the differential rostro-caudal patterning of angiotensinogen (AoGen) and AT1A receptor mRNAs in the rat SFO using specific and validated oligodeoxynucleotide probes for in situ hybridization. Highest levels of AoGen-specific gene expression were observed in the rostral region of the SFO with gradually decreasing intensity towards the caudal region of this sensory circumventricular organ lacking blood-brain barrier function. AoGen-related hybridization signals proved to be specifically prominent above cells in lateral aspects of the SFO, surrounding septal venules. Maximal expression of the AT1A receptor-specific gene, on the other hand, could be detected in the neuron-enriched, ventro-medial core region and dorsal annulus of the SFO, with low-intensity hybridization signals in its rostral and caudal parts. Water deprivation for 48 h, leading to extracellular hypertonic hypovolemia with elevated circulating AngII concentrations within the physiological range, caused a significant increase in AoGen-specific hybridization signals in the rostral and medial SFO regions. AT1A receptor gene expression and AngII receptor binding were markedly stimulated in the medial and caudal regions of the SFO (core and annulus) as compared to euhydrated animals. These data indicate, that mild dehydration differentially up-regulates AoGen- and AT1A receptor-specific mRNA formation as well as AT1 receptor binding in distinct regions of the SFO, and supports the involvement of different cellular subgroups in the expression of two major components of the central nervous renin-angiotensin system in this sensory circumventricular organ.     

Peptide and acetylcholine action on neurones of the cat subfornical organ

This paper presents evidence for an IgG1 allotype detected by a sheep antibovine serum. The character which appears to be inherited in a simple Mendelian way has been named G1a1.     

Short exposure to endotoxin increases the constriction induced by angiotensin II in rat aorta

The contraction elicited by angiotensin II (ANG II) was studied by using standard isometric tension techniques in aortic rings exposed for 1 h to 1 or 10 micrograms/ml Escherichia coli lipopolysaccharide endotoxin (LPS). This contraction was 18 and 71% greater for the two doses of LPS, respectively, than in unexposed control rings. In endothelium-denuded rings, the LPS-induced increase in contraction in response to ANG II was completely abolished. Because the contraction induced by ANG II is modulated by the simultaneous release of prostaglandins, we tested the hypothesis that LPS interferes with this modulation. We found that the LPS-induced increase in contraction to ANG II was inhibited in the presence of the cyclooxygenase inhibitor indomethacin (10(-5) M) or the prostaglandin H2/thromboxane A2-receptor antagonist SQ-29548 (2 x 10(-7) M). Conversely, the LPS-induced increase in contraction in response to ANG II was not inhibited by the presence of dexamethasone (10(-6) M), which inhibits new protein synthesis. In addition, there was no loss of vasodilator response to the endothelium-dependent receptor agonist acetylcholine (10(-8)-10(-4) M) or in the constrictor responses to norepinephrine (10(-9)-10(-5) M) and KCl (20-100 mM). We conclude that short exposure to LPS produces a specific increase in the constrictor response to ANG II via mechanisms mediated by prostaglandin H2/thromboxane A2. This effect could be a LPS-induced shift in favor of constrictor prostanoids in the balance of dilator/constrictor prostanoids, the release of which is associated with stimulation by ANG II.     

Angiotensin I modulates Ca-transport systems in the rat heart through angiotensin II

The objective of this study was to determine the effect of angiotensin I (Ang I) treatment in vivo on two major Ca-transport systems-the L-type voltage dependent calcium channel (L-VDCC) and the Na/Ca exchanger in rat heart. For our experiments we used four groups of rats, treated differently with saline, Ang I, the ACE inhibitor enalapril and/or combination of both for 6 days, every 24 h. We observed an increase in the activity, and also in mRNA expression of the Na/Ca exchanger, after repeated administration of Ang I in vivo. The maximal binding capacity of Ca-antagonist PN 200-110, which binds to the alpha 1 subunit of the L-VDCC was elevated from 0.8-1.85 pg/mg protein. mRNA expression of the voltage-dependent calcium channels of L-type system was also upregulated by Ang I administration, but not when enalapril was applied simultaneously with Ang I. These results demonstrate that in vivo application of the Ang I significantly modulates not only the activity, but also expression of the Na/Ca exchanger and the L-VDCC in rat hearts through angiotensin II (Ang II). Since in the in vitro experiments on the isolated cardiomyocytes, Ang II (100 nM) increased the calcium uptake after depolarization, and the AT1 receptor agonist losartan prevented this increase, we assume that this regulation might involve the AT1 receptors.     

Effects of angiotensin II and its blockers Sar1-Ile8-angiotensin II and DuP 753 on drinking in ducks in relation to properties of subfornical organ neurons

Properties of systemically applied angiotensin II in stimulating water intake of normally hydrated ducks were studied and the results compared with properties of angiotensin II-responsive neurons of the subfornical organ which are considered as targets for circulating angiotensin II acting as a dipsogen. Following intravenous infusion of hypertonic saline (2000 mosmol.kg-1 at 0.3 ml.min-1 for 1 h), intravenous infusion of 0.3 ml.min-1 isotonic saline with angiotensin II (200 ng.min-1), starting 1 h later, stimulated drinking in each case at an angiotensin II plasma level of about 1400 pg.ml-1. Without hypertonic priming, the same angiotensin II infusion did not stimulate drinking in each experiment; however, if effective, repeated infusions of ANGII induced stable dipsogenic responses. Angiotensin II infusions did not alter plasma levels of antidiuretic hormone. Sar1-Ile8-angiotensin II, a non-selective angiotensin II antagonist, acted weakly as a partial agonist when infused at a dose 200-fold higher than angiotensin II and effectively blocked the dipsogenic action of angiotensin II; this corresponds to the inhibition of angiotensin II-induced excitation by Sar1-Ile8-angiotensin II observed in duck subfornical organ neurons. DuP 753 (losartan), an angiotensin II antagonist specifically blocking AT1 receptors in mammals, had equivocal effects on angiotensin II-induced drinking in ducks at rates 50- and 200-fold higher than angiotensin II, which corresponds to the weak inhibitory action of this compound on angiotensin II-induced neuronal excitation in the duck SFO. Blood pressure was only marginally elevated by the applied angiotensin II dose and Sar1-Ile8-angiotensin II had no effect.     

Pharmacological analysis of local anaesthetic tolycaine-induced convulsions by modification of monoamines in rat brain

The effects of a local anaesthetic, tolycaine, on brain monoamine levels were investigated during the convulsive process in rats. The influence of central monoamine modifications on tolycaine-induced convulsions was also examined. Tolycaine (140 mg/kg, intraperitoneally) produced a significant elevation of noradrenaline and 5-hydroxytryptamine levels in all brain regions in the convulsive state from the levels in the non-convulsive state. Their levels returned to normal during the postconvulsive state. Dopamine levels were depleted in the cerebral cortex, the striatum, and the ponsmedulla oblongata during the convulsive process and increased in the cerebellum. Pretreatment with alpha-methyl-p-tyrosine, which depletes brain catecholamine, suppresses the tolycaine-induced convulsions, as shown by a decrease in the incidence; L-3,4-dihydroxyphenylalanine and bis-(1-methyl-4-homopiperazinyl-thiocarbonyl)-disulfide, which increase brain catecholamine, intensified the convulsions, as shown by shortening of the latency and increase in the mortality. Antagonists of beta-adrenergic and dopamine receptors, such as propranolol, chlorpromazine and pimozide, markedly suppressed the convulsions, but an antagonist of alpha-adrenergic receptor, phenoxybenzamine, had no effect. Furthermore, 5-hydroxytryptophan, which increases brain 5-hydroxytryptamine, suppressed the convulsions, and DL-p-chlorophenylalanine, which depletes brain 5-hydroxytryptamine, intensified them. Antagonists of 5-hydroxytryptamine receptor, methysergide and methiothepin, suppressed the convulsions. These results suggest that brain noradrenaline and 5-hydroxytryptamine are major regulators in the tolycaine-induced convulsive process and that central catecholaminergic neurones act in a stimulatory way on the tolycaine-induced convulsions, while serotonergic neurones act suppressively.     

Localization of AT2 angiotensin II receptor gene expression in rat brain by in situ hybridization histochemistry

To localize the gene expression of AT2 angiotensin II receptors in rat brain we performed in situ hybridization histochemistry using 35S-labeled antisense riboprobes. The AT2 receptor mRNA expression pattern was compared in consecutive brain sections, from 2 week old rats, with the receptor expression by means of [125I]Sar1-ANG II binding and displacement with AT2 selective ligands followed by autoradiography. Expression of AT2 receptor mRNA was found in several thalamic nuclei (ventral posterolateral, mediodorsal, central medial, paracentral, and paraventricular), the medial geniculate nuclei, the nucleus of the optic tract, the subthalamic nucleus, the interposed nucleus of the cerebellum, and in the inferior olive. In these areas the AT2 receptor gene expression corresponds well with [125I]Sar1-ANG II binding. In addition, AT2 receptor mRNA expression was found in the red nucleus where no [125I]Sar1-ANG II binding was present. No significant hybridization of the AT2 receptor antisense probe was found in septal nuclei, the locus coeruleus, the dorsolateral geniculate nucleus, or the cerebellar cortex, areas rich in [125I]Sar1-ANG II binding. Our results indicate that some brain regions may be involved in AT2 receptor formation, transporting the receptor protein to other brain areas. However, in most structures, both the formation and expression of receptors occur, suggesting the existence of local AT2 receptor circuits, or that of AT2 autoreceptors. Other structures express only the receptor protein, indicating that these AT2 receptors are produced elsewhere. Our present data are the basis for further studies on the clarification of AT2 receptor pathways in the brain.     

Nigrostriatally induced motor reactions in the rat. I. Rotational behavior and posture asymetry after intracerebral injection of apomorphine and dopamine

Using a rotameter described by Ungrstedt, the influence of pretreatment with 6-hydroxy-dopamine and transections of the Capsula interna on the asymmetry of the animal's poise and movement following systemic and intracerebral administration of dopamine and apomorphine was studied. After lesion of the nigrostriatal tract, i.p. administered apomorphine caused the animals to rotate towards the damaged side. After injection of apomorphine in the Nucleus caudatoputamen of healthy animals, initial rotations towards the injection side with subsequent opposite rotation were observed, whereas dopamine injected into the Nucleus caudatoputamen and the Substantia nigra initiated rotations in contralateral direction only. Pretreatment with haloperidole nullified the effect of apomorphine. The results have proved the effectiveness both in the Nucleus caudatoputamen and the Substantia nigra of drugs stimulating the dopamine receptors. With intact rats, the two sides of the nigrostriatal system are functionally asymmetric, which is reflected by the quantitative differences of responses following stimulation of dopamine-sensitive receptors and the individually different preference of one rotational direction. These individual behavioural patterns are modified by experimental influences.     

Characterization of phosphoinositide hydrolysis products induced by hexachlorocyclohexane isomers in rat brain cortex

Water-soluble inositol metabolites were separated by anion-exchange chromatography in order to determine whether or not gamma-hexachlorocyclohexane (gamma-HCH, lindane) and related compounds affect phosphatidylinositol hydrolysis in rat brain cortex slices. Hydrolysis was increased by delta- and gamma-HCH, while alpha- and beta-HCH were inactive. Muscarinic receptor stimulation of rat cortical slices with carbachol increases inositol phosphates formation. The combined effect of carbachol and the hexachlorocyclohexane isomers together were approximately equal to the sum of the effect of each one separately. The results suggest that lindane stimulates phosphoinositide phospholipase C and/or inhibits the phosphatases implicated in dephosphorylation of inositol phosphates.     

Differential effects of angiotensin II on the activities of suprachiasmatic neurons in rat brain slices

The effects of angiotensin II (AII) on the firing rates of suprachiasmatic neurons were determined in rat brain slices. AII in pmol ranges stimulated 25% and inhibited another 25% of 52 irregular firing neurons, while it stimulated 23% and inhibited 4% of 30 regular firing neurons. Three "oscillating" neurons whose firing rates oscillated with rather constant amplitudes and periods were recorded. AII induced the occurrence of oscillation in one unit and modulated the oscillation amplitude of the other two. Pretreatment with saralasin, an AII antagonist, effectively blocked (100%) the actions of AII (n = 5). The present findings suggest that AII may act as an important mediator in the suprachiasmatic nucleus and its mode of action may be variable in different neurons.