Central endogenous vasopressin induced by central salt-loading participates in body fluid homeostasis through modulatory effects on neurones of the paraventricular nucleus in conscious rats.

Peripherally secreted arginine vasopressin (AVP) plays a role in controlling body fluid homeostasis, and central endogenous AVP acts as a neurotransmitter or neuromodulator. The limbic system, which appears to exert an inhibitory effect on the endocrine hypothalamus, is also innervated by fibres that contain AVP. We examined whether central endogenous AVP is also involved in the control of body fluid homeostasis. To explore this possibility, we examined neuronal activity in the paraventricular nucleus of the hypothalamus (PVN), periventricular parts of the PVN and limbic brain areas, as well as AVP mRNA expression in the PVN and the peripheral secretion of AVP after central salt-loading in rats that had been pretreated i.c.v. with the AVP V(1) receptor antagonist OPC-21268. Neuronal activity in the PVN evaluated in terms of Fos-like immunoreactivity (FLI), especially in the parvocellular subdivisions, was suppressed. On the other hand, FLI was enhanced in the lateral septum, the bed nucleus of the stria terminalis and the anterior hypothalamic area. Similarly, AVP mRNA expression was enhanced in the magnocellular subnucleus of the PVN, despite the lack of a significant difference in the peripheral AVP level between OPC-21268- and vehicle-pretreated groups. We recorded renal sympathetic nerve activity (RSNA) as sympathetic nerve outflow during central salt-loading. The suppression of RSNA was significantly attenuated by i.c.v. pretreatment with OPC-21268. These results suggest that the suppression of RSNA during central salt-loading might be the result of a decrease in neuronal activity in the parvocellular subdivisions of the PVN via the inhibitory action of central endogenous AVP. The parvocellular and magnocellular neurones in the PVN might show different responses to central salt-loading to maintain body fluid homeostasis as a result of the modulatory role of central endogenous AVP.

Neuronal nitric oxide strongly suppresses sympathetic outflow in high-salt Dahl rats.

OBJECTIVE: To investigate the effects of a selective inhibitor of neuronal nitric oxide synthase (nNOS), 7-nitroindazole, on peripheral sympathetic outflow in Dahl rats. DESIGN AND METHODS: Dahl salt-sensitive and salt-resistant rats were fed either a regular-salt (0.4% NaCl) or a high-salt (8% NaCl) diet for 4 weeks. In chronically instrumented conscious rats, renal sympathetic nerve activity (RSNA) was measured in both baroreceptor-loaded and baroreceptor-unloaded states. The baroreceptor unload was performed by decreasing arterial pressure with occlusion of the inferior vena cava. RESULTS: 7-Nitroindazole (307 micromol/kg intraperitoneally) increased resting RSNA from 24 +/- 3% to 38 +/- 6% with an increase in mean arterial pressure of 15 +/- 3 mmHg, and increased baroreceptor-unloaded RSNA from 100% to 278 +/- 16% in salt-sensitive Dahl rats receiving a high-salt diet However, 7-nitroindazole did not increase resting RSNA, but did increase baroreceptor-unloaded RSNA from 100% to 179 +/- 15%, 177 +/- 15%, and 133 +/- 4% in salt-sensitive Dahl rats receiving a regular-salt diet, salt-resistant Dahl rats receiving a high-salt diet, and salt-resistant Dahl rats receiving a regular-salt diet, respectively. The high-salt diet significantly increased the baroreceptor-unloaded RSNA more than the regular-salt diet did, in both salt-sensitive and salt-resistant rats. After administration of the vehicle for 7-nitroindazole (peanut oil), L-arginine (100 micromol/kg per min for 10 min) decreased both resting and baroreceptor-unloaded RSNA, whereas after pretreatment with 7-nitroindazole, the L-arginine-induced suppression was reversed, in Dahl salt-sensitive rats receiving a high-salt diet. CONCLUSIONS: Neuronal nitric oxide may suppress the sympathetic discharge generated before baroreflex-mediated inhibition in all rats. This neuronal nitric oxide-mediated suppression was enhanced by the salt load in both salt-resistant and salt-sensitive Dahl rats. Finally, the neuronal nitric oxide-mediated suppression in tonic peripheral sympathetic outflow may be greatly enhanced in salt-sensitive hypertension.

Expression and electrophysiological function of actin in chick cerebellar neurons.

Among several monoclonal antibodies obtained by immunizing Balb/c mice with cerebellar synaptic membrane fractions from E20 chick embryos, the antibody, named M35, suppressed Ca-spikes in immature cultured chick cerebellar neurons. M35 immunoprecipitated 43 kDa protein from a 125I-labeled embryonic crude cerebellar membrane fraction. Immunohistochemically, the M35 antigen was expressed most intensively in Purkinje cells, but its expression was limited to highly motile structures at developmental neuronal remodeling. Electrophysiologically, M35 facilitated current responses to AMPA and inhibited the responses to GABA in cultured cerebellar Purkinje neurons. The several peptides derived from the affinity-purified 43 kDa protein were found to have homologous amino acid sequences to non-muscle actins. These results suggest that the antigen recognized by M35 may play an essential role probably as membrane ion channels modulating synaptic functions in not only the development and growth but also the neuronal activity of chick cerebellar Purkinje cells.

Detection of novel carbohydrate binding activity of interleukin-1.

Tamm-Horsfall glycoprotein (THGP) and the oligosaccharide fraction liberated from THGP by hydrazinolysis inhibited tetanus toxoid-induced T cell proliferation. Intact THGP showed approximately 100-fold more inhibitory activity than the free oligosaccharides. After fractionating the oligosaccharides by anion-exchange column chromatography, the inhibitory activity could be detected in a sialidase-resistant acidic oligosaccharide fraction (fraction AR). The inhibitory activity of fraction AR was not observed when the fraction was added to the T cell culture medium 24 h after the addition of tetanus toxoid. Increased concentration of interleukin (IL) 1beta and decreased concentration of IL-2 were observed in the T cell culture medium after the addition of fraction AR. The oligosaccharides in fraction AR also inhibited the growth of an IL-1-dependent cell line, D10-G4. These results strongly suggested that the oligosaccharides in fraction AR bind to IL-1beta and suppress its cytokine activity. IL-1beta actually bound to the fraction AR immobilized on an amino-bonded thin layer plate. Fractionation of the oligosaccharides indicated that only oligosaccharides containing an N-acetylgalactosamine residue and a sulfate residue bound specifically to IL-1beta. Removal of either the sulfate residue or the N-acetylgalactosamine residue from the oligosaccharides abolished both the proliferation-inhibition and IL-1beta binding activities. Since IL-1beta did not bind to thyroid-stimulating hormone, which has the sulfate group at C-4 of the N-acetylgalactosamine residue in its N-linked sugar chains, the binding of IL-1beta toward oligosaccharides in fraction AR was considered to be highly specific.