PTH-receptors regulate norepinephrine release in human heart and kidney

Recent data suggests that chronic renal failure and hyperparathyroidism are associated with sympathetic overactivity. Since peptide hormones are known to modulate norepinephrine (NE) release by activating prejunctional receptors, this study investigates whether parathyroid hormone fragment (1-34) (hPTH(1-34)) increases neuronal NE release in human heart and kidney. Using specific PTH-receptor agonists and antagonists, this study furthermore highlights functional differences between PTH1 and PTH2 receptors. Human atrial and renal tissues were incubated with [(3)H]-NE and superfused. Three electrical stimulations (5Hz, 1min) induced a stable [(3)H]-NE release which was taken as an index of endogenous NE release. RT-PCR with specific primers for PTH1- and PTH2-receptor was performed in heart and kidney. hPTH(1-34) (0.01-0.1μmol/L) and a stable analog of its second messenger cAMP (8-bromo-cAMP) increased [(3)H]-NE release in human atria. This facilitatory effect of PTH was also observed in human renal cortex. The PTH1-receptor antagonist (D-Trp(12), Tyr(34))-pTH-(7-34) (0.5μmol/L) abolished the effect of hPTH(1-34). This data was verified using isolated perfused mouse kidneys. Tuberoinfundibular peptide of 39 residues (TIP-39) (0.1nmol/L-0.1μmol/L) decreased [(3)H]-NE release in atria. PTH1- and PTH2-receptor expressions were demonstrated in human heart and kidney. Moreover, a splice variant of the PTH2-receptor was detected in human kidney. In conclusion, PTH is able to facilitate NE release in human atria and renal cortex by activation of PTH1-receptors. The highly increased PTH levels that can be observed in chronic renal failure might be one contributor for the elevated sympathetic nerve activity and the associated cardiovascular mortality in patients with end stage renal disease.     

The effect of age and norepinephrine on renin release by rat kidney slices in vitro.

Basal renin release by rat kidney slices decreases with age in rats of SHR and Sprague-Dawley strains. In contrast, Kyoto-Wistar rats (from which SHR are derived) demonstrate no decrease in renin release with age. The decline in basal renin release observed in SHR occurs at a time when the animal develops hypertension. However, the ability of renin release to respond to stimuli, such as norepinephrine, is enhanced at the time of declining basal renin release and developing hypertension.     

Support for a role for feedback regulation of norepinephrine release

There is abundant evidence that norepinephrine (NE) and other sympathomimetic amines with alpha-adrenoceptor activity inhibit the electrically evoked release of NE, whereas phenoxybenzamine and other alpha-adrenergic blocking agents enhance the electrically evoked release of NE. The physiological relevance of these observations, however, is disputed. The intent of this paper is to show that alpha-adrenergic blocking agents generally enhance transmitter output on nerve stimulation, but that some are more selective for presynaptic than for postsynaptic alpha receptors. Suggestions are made to account for the modulation of NE release as evoked by a single pulse.     

Regulation of hypoxia-induced release of corticotropin-releasing factor in the rat hypothalamus by norepinephrine

Corticotropin-releasing factor (CRF) peptide release was activated by hypoxia in the rat hypothalamus. The mechanisms, however, of the hypoxia-induced CRF release remains unclear. In this study, we demonstrated that the norepinephrine (NE) and its receptors in the paraventricular nucleus (PVN) mediated the CRF release in a simulated altitude hypoxia. When rats were exposed to 5 or 7 km altitude of hypoxia for a short or long term: (1) NE levels in the PVN and the CeA, using the HPLC analysis, were intensity and time course dependently increased, but the increase in the PVN were potential than in the CeA. Restraint-induced NE increase was much higher in both the PVN and the CeA, compared with hypoxia-induced response. (2) Hypoxia and restraint significantly enhanced CRF release in the ME and the PVN but not in the CeA, through RIA assay, which result in stimulating corticosterone secretion. (3) Hypoxia-induced CRF release was reversed by an injection of prazosin (i.c.v.), an alpha-1 adrenoceptor antagonist, while administration of yohimbine (i.c.v.), an alpha-2 receptor antagonist, facilitated further CRF release. These data suggested that hypoxia induced NE activation centrally, via alpha-1 and -2 receptors, leading to improving hypothalamic CRF release, which in turn stimulated pituitary and adrenal cortex. Restraint presented much potential action on NE activation than hypoxia.     

Increases in norepinephrine release and ovarian cyst formation during ageing in the rat

Background  

Depletion of ovarian follicles is associated with the end of reproductive function in ageing females. Recently, it has been described that this process parallels increases in the concentration of norepinephrine (NE) in the rat ovary. In sexually mature rats, experimentally-induced increases in the sympathetic tone of the ovary is causally related to ovarian cyst formation and deranged follicular development. Thus, there is a possibility that increased ovarian NE concentrations represent changes in the activity of sympathetic nerves, which consequently participate in the process of ovarian cyst formation observed during ageing in the human and experimental animal models.     

Effects of atrial natriuretic factor on norepinephrine release in the rat hypothalamus.

The effects of atrial natriuretic factor on the mechanisms involved in norepinephrine release were studied 'in vitro' in slices of Wistar rat hypothalamus. Atrial natriuretic factor (10, 50 and 100 nM) decreased spontaneous [3H]norepinephrine secretion in a concentration dependent way. In addition, the peptide (10 nM) also reduced acetylcholine induced output of norepinephrine. The atrial factor (10 nM) was unable to alter the amine secretion when the incubation medium was deprived of calcium or when a calcium channel blocker such as diltiazem (100 microM) was added. In conclusion, atrial natriuretic factor reduced both spontaneous and acetylcholine evoked [3H]norepinephrine release in the rat hypothalamus. These findings suggest that the atrial natriuretic factor may alter catecholamine secretion by modifying the calcium available for the exocytotic process of catecholamine output.     

Hepoxilin A3 blocks the release of norepinephrine from rat hippocampal slices

Hepoxilin A3 was previously shown to display neuromodulatory actions on rat hippocampal CA1 neurons, with hyperpolarization of the membrane potential, an increase in the amplitude and duration of the post-spike train after hyperpolarization and an increase in the inhibitory post synaptic potential. The present report describes new biochemical evidence of a presynaptic action of hepoxilin A3 in rat hippocampal slices prelabeled with [3H]-norepinephrine. Hepoxilin A3 on its own had a marginal effect on the release of label, but blocked release which was induced by 4-aminopyridine (4-AP). Prostaglandin E2 also behaved in a similar way. These results demonstrate that hepoxilins modulate neurotransmission in the mammalian CNS through both pre- and postsynaptic actions.     

Angiotensin II-induced release of oxytocin: interaction with norepinephrine and role in lactation

These studies examined the receptors involved in angiotensin II (Ang II) stimulated secretion of systemic oxytocin (OT) and the role of this peptide in release of OT during suckling. Plasma OT concentrations were measured following intracerebroventricular (icv) injection of vehicle, Ang II, or Ang II following pretreatment with a selective AT1 (Losartan) or AT2 (PD 123319) receptor antagonist. Furthermore, we measured Ang II-induced OT release during central alpha-adrenergic receptor blockade (phentolamine). Finally, plasma OT concentrations before and during suckling were evaluated following central administration of Ang II receptor antagonists. The increase in systemic OT following central Ang II was abolished by AT1 receptor blockade and inhibited by the AT2 receptor antagonist. Furthermore, pretreatment with phentolamine significantly diminished systemic OT release in response to icv Ang II. Finally, central Ang II receptor blockade did not alter the increase in circulating OT during suckling. These data demonstrate that Ang II evoked OT release is mediated through activation of both AT1 and AT2 receptors and suggest that a component of Ang II-induced OT stimulation is due to norepinephrine release. Furthermore, central angiotensin systems do not have a direct role in stimulating OT release during suckling.     

Endothelin-induced modulation of neuropeptide Y and norepinephrine release from the rat mesenteric bed

The effect of three endothelin (ET) agonists [ET-1, ET-3, and sarafotoxin (STX6C)] on the nerve stimulation-induced release of norepinephrine (NE) and neuropeptide Y-immunoreactive compounds (NPY-ir) from the perfused mesenteric arterial bed of the rat as well as the effect on perfusion pressure were examined. ET-1, ET-3, and STX6C all produced a significant, concentration-dependent decrease in the evoked release of NPY-ir but had no effect on the release of NE. In contrast, all three ETs potentiated the nerve stimulation-induced increase in perfusion pressure. The inhibition of nerve stimulation-induced NPY-ir release by ET-1 was significantly blocked by the ET(A)/ET(B) antagonist PD-142893 and the ET(B) antagonist RES-701-1 but not by the ET(A) antagonist BQ-123. The potentiation of the nerve stimulation-induced increase in perfusion pressure by ET-1 was significantly blocked by PD-142893 and BQ-123 and attenuated by RES-701-1. Prior exposure of the preparation to indomethacin or meclofenamate failed to alter the attenuation of the evoked release of NPY-ir or the potentiation of the increase in perfusion pressure produced by ET-1 or ET-3. These results are consistent with the idea that sympathetic cotransmitters can be preferentially modulated by paracrine mediators at the vascular neuroeffector junction.     

NMDA-induced acetylcholine release in mouse striatum: role of NO synthase isoforms

Striatal cholinergic interneurons are stimulated by glutamatergic inputs from thalamus and cortex via NMDA receptors. The present microdialysis study was designed to characterize the role of nitric oxide (NO) in this process and to identify the NO synthase (NOS) isoform responsible for this effect. For this purpose, we studied the effects of NMDA and 3-morpholino sydnonimine (SIN-1) perfusions on the release of acetylcholine (ACh) in mouse striatum. In wild-type C57/Bl6 mice, perfusion of NMDA (100 micro m) induced a two-fold stimulation of ACh release. This effect was attenuated in mice lacking endothelial NOS but was completely absent in mice lacking neuronal NOS. Local perfusion of SIN-1 (300 micro m), an NO donor, increased ACh release by more than two-fold in all three mouse lines. We conclude that NO synthesized by neuronal NOS provides a nitrergic link in the glutamatergic stimulation of striatal cholinergic interneurons.     

Stimulatory effect of somatostatin on norepinephrine release from rat brain cortex slices

The effect of somatostatin (SRIF) on norepinephrine (NE) release from the brain tissue was determined on the superfused rat cerebral cortex slices preloaded with ³HNE. SRIF (0.38 μM–1.53 μM) was found to stimulate dose-dependently tritium (³H) overflow evoked electrically by 30%—116% although SRIF did not affect on the spontaneous ³H overflow. SRIF at the concentrations which exhibited the stimulatory effect inhibited scarecely the uptake of ³HNE by cortex slices, while the reference drug, cocaine (50 μM, 10 μM) markedly depressed the uptake. The stimulatory effect of SRIF was not reduced by phentolamine (3.14 μM), α-adrenoceptor blocker, which increased the evoked ³H overflow from the slices itself. These results suggest that SRIF does not produce its stimulatory effect by inhibiting the NE reuptake mechanisms or by interacting with the presynaptic α-adrenoceptors. Elevating of Ca²⁺ concentrations from 0.75 mM to 2.25 mM in the superfusion fluid reduced the stimulatory effect of SRIF. It is possible that SRIF stimulates NE release by facilitating the availability of Ca²⁺ for the release mechanisms.     

Volume-regulatory potassium release from isolated perfused rat kidney

The present study has been performed to test for cell volume regulatory potassium release from the isolated perfused rat kidney exposed to hypotonic perfusate and for its sensitivity to potassium channel blocker barium and calcium channel blocker verapamil. Replacement of 25 mmol/l NaCl with 50 mmol/l mannitol has little effect on effluent potassium activity, whereas subsequent omission of mannitol from the perfusate leads to a transient increase of effluent potassium activity, reflecting volume regulatory potassium release. Barium (1 mmol/l) leads to a marked transient decrease of effluent potassium activity, pointing to net cellular uptake of potassium. Verapamil (1 mumol/l) leads to a slight decrease of effluent potassium activity. Both barium and verapamil virtually abolish the rapid, transient increase of effluent potassium activity upon exposure to hypotonic perfusates. Thus, the substances either block or markedly retard volume regulatory potassium release. The apparent renal vascular resistance is transiently increased by exposure to hypotonic perfusates and by barium, but is reduced by verapamil. Cell volume regulation of isolated perfused mouse straight proximal tubules is retarded but not abolished by verapamil (0.1 mmol/l). In conclusion, cellular potassium release from rat kidney can be determined by continuous measurement of effluent potassium activity. The volume regulatory potassium release and cell volume regulation are impaired by both barium and verapamil. The persisting cell volume regulation could be due either to slow potassium release and/or some mechanism independent of potassium.     

Modulation of GABA-augmented norepinephrine release in female rat brain slices by opioids and adenosine

GABA_A receptor activation augments electrically-stimulated release of norepinephrine (NE) from rat brain slices. Because this effect is not observed in synaptoneurosomes, GABA probably acts on inhibitory interneurons to disinhibit NE release. To determine whether opioids or adenosine influence GABA-augmented NE release, hypothalamic and cortical slices from female rats were superfused with GABA or vehicle in the presence and absence of 10 M morphine or 100 M adenosine. GABA augments [³H]NE release in the cortex and hypothalamus. Morphine alone has no effect on [³H]NE release, but attenuates GABA augmentation of [³H]NE release in both brain regions. Adenosine alone modestly inhibits [³H]NE release in the cortex, but not in the hypothalamus. Adenosine inhibits GABA-augmented [³H]NE release in both brain regions. The general protein kinase inhibitor H-7, augments [³H]NE release in both brain regions and may have additive effects with GABA in cortical slices. These results implicate opioid and adenosine interneurons and possibly protein kinases in regulating GABAergic influences on NE transmission.     

Alpha-2 adrenergic regulation of norepinephrine release in the rat submandibular gland as measured by HPLC-EC

In many tissues, norepinephrine appears to inhibit its own release through an interaction at alpha adrenergic receptors. We have developed an assay for measuring the release of endogenous norepinephrine based on HPLC and have studied the regulation of release in the rat submandibular gland by alpha adrenergic antagonists. The method uses electrochemical detection to quantitate norepinephrine released from tissue slices and does not require preloading of the tissue with [3H]norepinephrine. Yohimbine, an alpha-2 adrenergic antagonist, potentiates by 50% the release caused by potassium induced depolarization with an EC50 of 0.14 microM. Prazosin, an alpha-1 antagonist, has a similar effect, but is less potent with an EC50 of 0.77 microM. Thus, the alpha adrenergic receptor mediating the regulation of norepinephrine release is of the alpha-2 subtype. The observed equal efficacies and lack of additivity of release potentiation by yohimbine and prazosin at maximal doses suggest that both drugs act at the same receptor. The five-fold difference in potency between prazosin and yohimbine is consistent with the recent observations indicating species differences between rodent and non-rodent alpha-2 adrenergic receptors.     

Nitrendipine-induced stimulation of renin release by the isolated perfused rat kidney

The direct effects of the organic calcium antagonist nitrendipine upon renin release were assessed using the isolated rat kidney perfused at constant pressure. This model circumvents the indirect actions of vasodilating agents by artificially maintaining perfusion pressure constant, thereby avoiding the hypotensive effects associated with the systemic administration of such agents. Renin release as assessed by radioimmunoassay was stimulated 2.6-fold upon the administration of 10(-6) M nitrendipine. Since this stimulation of renin release occurred in the absence of any alteration in perfusion pressure, we conclude that it represents a direct action of nitrendipine. This finding is in support of the current hypothesis concerning the inverse relationship between cytosolic Ca2+ and renin secretory rate, and suggests that Ca entry into the juxtaglomerular cells of the juxtaglomerular apparatus is sensitive to blockade by organic calcium antagonists such as nitrendipine.     

Insulin blockade of norepinephrine tachyphylaxis in the perfused kidney

Tachyphylaxis to norepinephrine (NOR) was determined in the rabbit kidney perfused with Krebs-Henseleit solution by using different calcium concentrations (2.5 mM; 5 mM; 12.5 mM) in the perfusate. The addition of insulin to the perfusion fluid causes a reversion of the tachyphylaxis which is seen at those Ca2+ concentrations. This effect is demonstrated mainly at 5 mM Ca2+. When kidneys were perfused with 12.5 mM calcium there was disappearance of NOR-mediated tachyphylaxis, both in the absence and in the presence of insulin. In this calcium concentration, insulin decreases vascular reactivity to NOR. These results suggest that insulin blockade of alpha adrenergic tachyphylaxis is a calcium-mediated effect which is thought to be due to an enhancement of calcium pumping inside the cells.     

Uptake and release of norepinephrine by rat brain tissue fractions prepared by ultrafiltration

Abstract— Isolated nerve endings were removed from crude homogenates of rat brain by Millipore filters of pore size 0-5-0-8 μm. Synaptosomes containing L-[³H]norepinephrine were incompletely removed from a non-ionic medium by 0-8 μm pore filters, but were nearly quantitatively removed from Krebs’medium, as demonstrated by density gradient subcellular distribution. In suspension, synaptosomes accumulated labelled norepinephrine. Catecholamine uptake was active; it was inhibited by ouabain, imipramine, cocaine, β-phenethylamine and amantadine. Bound norepinephrine was released by a high concentration of K⁺. Nerve endings trapped on ultrafilters behaved very similarly to synaptosomes suspended in Krebs’medium by actively accumulating norepinephrine and serotonin; they also possessed monoamine oxidase activity and norepinephrine was released from them by increased concentrations of K⁺. Ultrafiltration provides a simple, rapid method of preparing metabolically active synaptosomes.