High Magnesium Solution Research Articles

High-magnesium exposure to bullfrog heart causes ST segment elevation.

Hypermagnesemia occurs in elderly people or patients with renal insufficiency after excessive ingestion of magnesium-containing laxatives. In addition to typical electrocardiogram (ECG) findings caused by conduction defects, changes in the ST segments and T waves are also observed in patients with severe hypermagnesemia. This suggested the involvement of similar pathophysiology to acute myocardial infarction, as we previously demonstrated using burn-induced subepicardial injury model in frog hearts. In the present study, by exposing the bullfrog heart to high-magnesium solution, we reproduced prominent ST segment changes in ECG as actually observed in patients with severe hypermagnesemia. In addition to the great increase in the T waves, the ECG showed a marked elevation of the ST segments and the cardiac action potential demonstrated a marked shift of the resting membrane potential to the depolarized side. High-magnesium exposure did not affect the abundance of Na+/K+-ATPase proteins. However, the pharmacological stimulation of Na+/K+-ATPase activity by insulin quickly retrieved the elevated ST segments in ECG. From these results, the functional blockade of Na+/K+-ATPase activity by magnesium ions was thought to be responsible for generating the potassium concentration gradient and the subsequent ST segment changes.

Synthesis of magnesium sulfate from seawater using alkaline industrial wastes, sulfuric acid, and organic solvents

ABSTRACTIn this study, we used three processes to synthesize magnesium sulfate from seawater. First, alkaline industrial wastes, cement kiln dust and paper sludge ash, were injected to the seawater to precipitate magnesium in the form of magnesium hydroxide (Mg(OH)2). Then, magnesium was eluted with a small amount of H2SO4 to make a high concentration magnesium solution. Finally, an organic solvent was added to precipitate magnesium sulfate (MgSO4). Over 90% of magnesium was recovered through the three processes. It is expected that 11.3 kg of magnesium sulfate (based on MgSO4 · 6H2O) can be synthesized from 1 ton of seawater.

Effects of riluzole on respiratory rhythm generation in the brainstem-spinal cord preparation from newborn rat

The persistent sodium channel is an important pacemaker component in rhythm generation. In the present study, we examined the effects of a persistent sodium channel blocker, riluzole on pre-inspiratory (Pre-I) and inspiratory neurons in the rostral medulla as well as on 4th cervical ventral root (C4)-inspiratory activity in brainstem-spinal cord preparations. Preparations were isolated from postnatal day 0–3 Wistar rats and were superfused with artificial cerebrospinal fluid, equilibrated with 95% O2 and 5% CO2, pH 7.4, at 25–26°C. The C4 inspiratory burst rate decreased in a dose-dependent manner (50–200μM) after 15min application of riluzole. Riluzole caused a strong reduction in the drive potential of Pre-I neurons but not of inspiratory neurons. After washout, C4 inspiratory burst gradually changed into an episodic pattern, in which one burst consisted of 3–9 short separate bursts. Riluzole also depressed the induction of repetitive firing induced by depolarizing stimulation. Under voltage clamp conditions, riluzole suppressed the negative-slope component of Pre-I neurons. Riluzole also depressed the intrinsic burst generation of Pre-I neurons in low calcium and high magnesium solution. Our findings indicate that the burst generation of Pre-I neurons is more sensitive than inspiratory burst generation to riluzole and thus suggested that persistent sodium channels have an important role in the burst generation of Pre-I neurons and are involved in the primary respiratory rhythm generation.

Physical compatibility of magnesium sulfate and sodium bicarbonate in a pharmacy-compounded hemofiltration solution

The physical compatibility of magnesium sulfate and sodium bicarbonate in a pharmacy-compounded hemofiltration solution was assessed. Two bicarbonate-buffered hemofiltration solutions (low- and high-magnesium formulations) were compounded in triplicate. The concentrations of magnesium (15 meq/L) and sodium bicarbonate (50 meq/L) in the high-magnesium formulation were chosen to be somewhat below the concentrations reported as being incompatible in a popular reference. The six hemofiltration bags were stored at 22-25 degrees C without protection from light for 48 hours. Physical compatibility was assessed by visual inspection and microscopy. The pH of the solutions was assayed 3-4 and 52-53 hours after compounding. Electrolyte and glucose concentrations of the solutions were assayed at 3-4 and 50-51 hours after preparation. No particulate matter was observed by visual or microscopic inspection in the compounded hemofiltration solutions at 48 hours. The mean +/- S.D. pH values of the low-magnesium solutions were 8.01 +/- 0.02 and 8.04 +/- 0.02 at 3-4 and 52-53 hours after compounding, respectively. The mean +/- S.D. pH values of the high-magnesium solutions were 7.96 +/- 0.02 and 7.98 +/- 0.01 at 3-4 and 52-53 hours after compounding, respectively. The electrolyte and glucose concentrations in the low- and high-magnesium solutions were similar 3-4 and 50-51 hours after preparation. Magnesium sulfate 1.5 meq/L and sodium bicarbonate 50 meq/L were physically compatible in a pharmacy-compounded hemofiltration solution for 48 hours when stored at 22-25 degrees C without protection from light.

Neurotoxicity of a Novel Local Anesthetic Agent, Ropivacaine: The Possible Roles of Bursts of Potential and Cytoplasmic Second Messenger

Ropivacaine has been shown to induce convulsion following overdose or accidental intravenous injection, but the mechanisms are poorly understood. Using an identifiable central neuron from giant African snail, the authors studied the mechanism of ropivacaine-elicited bursts of potential and explored the possible mechanisms of ropivacaine-induced neurotoxicity. Ropivacaine action on a central neuron (RP4) of the giant African snail (Achatina fulica Ferussac) was recorded by conventional electrophysiologic technique. Interactions between ropivacaine and prazosin, propranolol, atropine, d-tubocurarine, calcium-free solution, H89, U73,122, neomycin, high-magnesium solution, and chelerythrine were also observed. The RP4 neuron showed spontaneous firing of action potentials. Extracellular application of ropivacaine (900 microM) reversibly elicited bursts of potential in the RP4 neuron. The bursts of potential elicited by ropivacaine were not blocked after administration of: (1) prazosin, propranolol, atropine, d-tubocurarine; (2) calcium-free solution; and (3) pretreatment with H89 or chelerythrine. The bursts of potential elicited by ropivacaine were blocked by pretreatment with U73122 (30 microM) or by adding neomycin (3.5 mM) or high-magnesium solution (30 mM). Ropivacaine reversibly elicited bursts of potential in the central snail neuron. The ropivacaine-elicited bursts of potential were associated with phospholipase C activity in the RP4 snail neuron. Our results suggest that ropivacaine-induced neurotoxicity is highly associated with phospholipase C activity and phospholipase C inhibitor may offer a novel therapeutic approach for managing local anesthetic-induced convulsion or other transient neurologic toxicity.

Effects of rolipram on induction of action potential bursts in central snail neurons

Effects of rolipram, a selective inhibitor of phosphodiesterases (PDE) IV, on induction of action potential bursts were studied pharmacologically on the RP4 central neuron of the giant African snail ( Achatina fulica Ferussac). Oscillations of membrane potential bursts were elicited by rolipram and forskolin. The bursts of potential elicited by rolipram were not inhibited after administration with (a) calcium-free solution, (b) high-magnesium solution (30 mM) or (c) U73122. However, the bursts of potential elicited by rolipram were inhibited by pretreatment with KT-5720 (10 μM). Voltage-clamp studies revealed that rolipram decreased the total inward current and steady-state outward currents of the RP4 neuron. The negative slope resistance (NSR) was not detectable in control or rolipram treated RP4 neurons. TEA elicited action potential bursts and an NSR at membrane potential between −50 mV and −30 mV. It is suggested that the bursts of potential elicited by rolipram were not due to (1) synaptic effects of neurotransmitters; (2) NSR of steady-state I– V curve; (3) phospholipase activity of the neuron. The rolipram-elicited bursts of potential were dependent on the phosphodiesterases inhibitory activity and the cAMP signaling pathway in the neuron.

Effects of procaine on a central neuron of the snail, Achatina fulica Ferussac

Effects of procaine on a central neuron (RP1) of the giant African snail ( Achatina fulica Ferussac) were studied pharmacologically. The RP1 neuron showed spontaneous firing of action potential. Extra-cellular application of procaine (10 mM) reversibly elicited bursts of potential. The bursts of potential elicited by procaine were not blocked after administration of (1) prazosin, propranolol, atropine, d-tubocurarine, (2) calcium-free solution, (3) ryanodine (4) pretreatment with KT-5720 or chelerythrine. The bursts of potential elicited by procaine were blocked by adding U73122 (10 μM) and the bursts of potential were decreased if physiological sodium ion was replaced with lithium ion or incubated with either neomycin (3.5 mM) or high magnesium solution (30 mM). Preatment with U73122 (10 μM) blocked the initiation of bursts of potential. Ruthenium red (100 μM) or caffeine (10 mM) facilitated the procaine-elicited bursts of potential. It is concluded that procaine reversibly elicits bursts of potential in the central snail neuron. This effect was not directly related to (1) the extra-cellular calcium ion fluxes, (2) the ryanodine sensitive calcium channels in the neuron, or (3) the PKC or PKA related messenger systems. The procaine-elicited bursts of potential were associated with the phospholipase activity and the calcium mobilization in the neuron.

Long-term potentiation of nicotinic synaptic transmission in rat superior cervical ganglia produced by phorbol ester and tetanic stimulation

The long-term potentiation of nicotinic synaptic transmission induced by both active phorbol ester (4β-phorbol-12,13-dibutyrate, PdBu) and tetanic trains of preganglionic stimulation was studied in single neurons of the superior cervical ganglion (SCG) of the rat using intracellular recording techniques. PdBu significantly increased the mean amplitude of both the unitary evoked fast excitatory postsynaptic potentials (EPSPs) and the fast excitatory postsynaptic currents (EPSCs) to 17.0±3.3 mV (control 8.4±1.9 mV, n=5) and 2.8±0.4 nA (control 0.8±0.1 nA, n=10), respectively. There was no significant change in either the resting membrane potential, input resistance, or the threshold for the initiation of an action potential. The response to exogenously applied acetylcholine (ACh) was also not changed following exposure to PdBu. In low-calcium, high-magnesium solutions, PdBu significantly increased the quantal content of EPSPs approximately threefold from a control of 0.9±0.2 ( n=5) to 2.6±0.6 ( n=5). The quantal content of EPSCs was also increased to 1.3±0.2 (control 0.5±0.1, n=10). PdBu increased the frequency of miniature EPSPs (mEPSPs) to 196±47% ( n=6) of control, while the amplitude, rise time, rate of rise, and decay of mEPSPs were not significantly changed. Tetanic stimulation significantly increased the amplitude of the unitary synaptic EPSPs and EPSCs without significantly changing the resting membrane potential, input resistance, threshold for initiation of an action potential, or the response to exogenously applied ACh. Tetanic stimulation significantly increased quantal content of EPSPs and EPSCs threefold. The results obtained with tetanically induced LTP are similar to the results obtained with phorbol ester-induced LTP in these ganglion neurons. These results suggest that both tetanically induced and phorbol ester-induced LTP, in the rat, share similar mechanisms which involve, at least in part, activation of PKC-dependent mechanisms to increase quantal release from sympathetic preganglionic axon terminals.

Neural integrity is essential for the propagation of colonic migrating motor complexes in the mouse.

The mechanisms that underlie the propagation of contractions along the colon are uncertain. We have examined whether spontaneous colonic migrating motor complexes (CMMCs) migrate through a region of muscle paralysis, or through a region where neural transmission was disrupted in the isolated mouse colon. Mouse colon was mounted in a separately perfused three-compartment organ bath and recordings of circular muscle tension were made. Drug application was restricted to the middle compartment. Application of nifedipine (1 micromol L(-1)), an l-type calcium channel antagonist, reduced the contraction amplitude by approximately 94%, without affecting the form of contractions in the proximal and distal compartments. Moreover, CMMCs appeared to remain temporally related in all compartments. In contrast, interruption of neural transmission in the middle compartment by either tetrodotoxin (1.6 micromol L(-1)), hexamethonium (500 micromol L(-1)) or a low-calcium, high-magnesium solution abolished CMMCs in this compartment; contractions recorded in the proximal and distal compartments became slower in frequency and were no longer synchronized. The experiments suggest that there may be more than one 'pacemaker' generating spontaneous CMMCs and that CMMCs can migrate through a region where there is minimal tension generation, but not through a region where neural integrity has been compromised.

Presynaptic nicotinic acetylcholine receptors in the myenteric plexus of guinea pig intestine

Presynaptic nicotinic acetylcholine receptors (nAChRs) were studied in myenteric plexus preparations from guinea pig ileum using intracellular electrophysiological methods. Microapplication of nicotine (1 mM) caused a biphasic depolarization in all AH neurons (n = 30) and in 36 of 49 S neurons. Cytisine (1 mM) caused fast depolarizations in S neurons and no response in AH neurons. Mecamylamine (10 microM) blocked all responses caused by nicotine and cytisine. TTX (0.3 microM) blocked slow excitatory synaptic potentials in S and AH neurons but had no effect on fast depolarizations caused by nicotine. Nicotine-induced slow depolarizations were reduced by TTX in two of twelve AH neurons (79% inhibition) and four of nine S neurons (90+/-12% inhibition). Slow nicotine-induced depolarizations in the remaining neurons were TTX resistant. TTX-resistant slow depolarizations were inhibited after neurokinin receptor 3 desensitization caused by senktide (0.1 microM); senktide desensitization inhibited the slow nicotine-induced depolarization by 81+/-5% and 63+/-15% in AH and S neurons, respectively. A low-calcium and high-magnesium solution blocked nicotine-induced slow depolarizations in AH neurons. In conclusion, presynaptic nAChRs mediate the release of substance P and/or neurokinin A to cause slow depolarizations of myenteric neurons.

Mesencephalic trigeminal neuron responses to γ-aminobutyric acid

Mesencephalic trigeminal neurons are primary sensory neurons which have cell somata located within the brain stem. In spite of the presence of synaptic terminals on and around the cell somata, applications of a variety of neurotransmitter substances in earlier studies have failed to demonstrate responses. Using intracellular recording in a brain slice preparation, we have observed prominent depolarizations and decreases in input resistance in response to applications of γ-aminobutyric acid (GABA) in most recorded mesencephalic trigeminal neurons. Those cells failing to respond were located deeply within the slice, and the low responsiveness was shown to be related to uptake of GABA in the slice. The responses were direct, since they remained during perfusion with a low calcium, high magnesium solution that blocks synaptic transmission. The responses were mimicked by the GABA A receptor agonist isoguvacine, and blocked by GABA A receptor antagonists. The GABA B receptor agonist baclofen evoked no changes in membrane potential or input resistance in neurons exhibiting depolarizations with GABA application. Tests of neuronal excitability during GABA applications indicated that the excitatory effects of the depolarization prevail over the depressant effects of the increase in membrane conductance. In situ hybridization histochemistry indicated that the GABA A receptors in Me5 cells are comprised of α 2, β 2 and γ 2 subunits.© 1997 Elsevier Science B.V. All rights reserved.

Angiotensin II and glutamate influence area postrema neurons in rat brain slices

The area postrema (AP) has been repeatedly implicated in cardiovascular regulation. Microinjection and single unit recording studies in vivo have suggested specific actions for angiotensin II (ANG) and glutamate (GLU) in controlling the excitability of AP neurons. The present study was therefore designed to examine the responsiveness of AP neurons to bath administration of these substances. Of the 133 AP neurons tested with ANG (10−8–10−6 M) 40% were excited, 13% inhibited and the remainder unresponsive. The excitatory effects of ANG on AP neurons were dose-dependent. Following blockade of synaptic transmission with a low calcium high magnesium solution excitatory responses were maintained in 12 of 15 cells tested. Pretreatment of slices with the AT1 receptor antagonist losartan blocked the excitatory effects of ANG in all cells (55) tested. The effects of GLU on AP neurons were also examined. Of the 71 AP cells tested, 40% were excited, 10% inhibited, 8% showed excitatory responses followed by periods of inhibition while the remaining cells were unaffected. Excitatory effects of GLU were maintained in all AP neurons (77) tested during perfusion with low calcium, high magnesium solutions. Similar responses to NMDA were observed in four of four cells tested, suggesting these GLU actions are mediated through NMDA receptors. These data demonstrate direct excitatory actions of ANG and GLU on AP neurons which are likely mediated through the AT1 and NMDA receptors, respectively.

Angiotensin II and glutamate influence area postrema neurons in rat brain slices

The area postrema (AP) has been repeatedly implicated in cardiovascular regulation. Microinjection and single unit recording studies in vivo have suggested specific actions for angiotensin II (ANG) and glutamate (GLU) in controlling the excitability of AP neurons. The present study was therefore designed to examine the responsiveness of AP neurons to bath administration of these substances. Of the 133 AP neurons tested with ANG (10 −8–10 −6 M) 40% were excited, 13% inhibited and the remainder unresponsive. The excitatory effects of ANG on AP neurons were dose-dependent. Following blockade of synaptic transmission with a low calcium high magnesium solution excitatory responses were maintained in 12 of 15 cells tested. Pretreatment of slices with the AT 1 receptor antagonist losartan blocked the excitatory effects of ANG in all cells ( 5 5 ) tested. The effects of GLU on AP neurons were also examined. Of the 71 AP cells tested, 40% were excited, 10% inhibited, 8% showed excitatory responses followed by periods of inhibition while the remaining cells were unaffected. Excitatory effects of GLU were maintained in all AP neurons ( 7 7 ) tested during perfusion with low calcium, high magnesium solutions. Similar responses to NMDA were observed in four of four cells tested, suggesting these GLU actions are mediated through NMDA receptors. These data demonstrate direct excitatory actions of ANG and GLU on AP neurons which are likely mediated through the AT 1 and NMDA receptors, respectively.

Capsaicin-sensitive afferent nerves activate submucosal secretomotor neurons in guinea pig ileum

This study examined whether capsaicin-sensitive sensory nerves regulate intestinal ion transport using both Ussing chamber and intracellular recording techniques in in vitro submucosal preparations from the guinea pig ileum. In Ussing chamber studies, serosal application of capsaicin (20 nM-20 microM) evoked a biphasic dose-dependent increase in short-circuit current (Isc) (maximal effective concentration 200 nM and 2 microM, respectively). In chloride-free buffer, capsaicin responses were significantly reduced. Capsaicin evoked little or no response when extrinsic sensory nerve fibers had been surgically removed and tetrodotoxin and low-calcium and high-magnesium solutions blocked responses to capsaicin. In epithelial preparations devoid of submucosal neurons, capsaicin had virtually no effect, suggesting that responses evoked by capsaicin-sensitive nerves result from activation of submucosal secretomotor neurons. Intracellular recordings from single submucosal neurons demonstrated that superfusion with capsaicin (2 microM) depolarized neurons with an associated decreased conductance. Depolarizations were completely desensitized when capsaicin was reapplied, but synaptic inputs were unaffected. This study suggests that capsaicin-sensitive nerves can regulate ion transport in the gastrointestinal tract by release of neurotransmitter(s) that activate submucosal secretomotor neurons.

Effects of Magnesium Ions on Detrusor Contraction in Rat

Contraction of smooth muscle including detrusor is initiated by an increase in the concentration of intracellular free calcium ions. Magnesium affects the translocation of calcium ions through the smooth muscle cell membrane. The effect of magnesium ions on detrusor contraction was investigated in vitro using the rat detrusor muscle. Detrusor contractility caused by intramural nerve stimulation increased in a magnesium-free solution but decreased in a high-magnesium solution (MgCl2 5 mM). Cumulative addition of MgCl2 suppressed the detrusor response induced by field stimulation dose-dependently, and 30 mM MgCl2 completely eliminated it. Mg ion also suppressed atropine-resistant detrusor contraction in a dose-dependent manner. After blocking nerve-mediated detrusor contractility with tetrodotoxin, Mg ions inhibited spontaneous detrusor contractions and reduced tonus of the muscle strips. Mg ions suppressed the detrusor contraction induced by carbachol and potassium. The effect of Mg ions on detrusor contraction was enhanced by verapamil but inhibited by Bay-K8644. These results suggest that Mg ions suppress detrusor muscle contractility through an inhibitory mechanism acting on calcium channels in the smooth muscle cell membrane.

Effects of magnesium ions on the contraction of the rat detrusor muscle

Effects of magnesium (Mg) ions on the contraction of the rat detrusor muscle induced by intramural nerve stimulation were investigated in vitro. 1. Frequency-response curve demonstrated that contractile responses increased in magnesium-free Krebs' solution and decreased in high magnesium solution (MgCl2 5 mM) in comparison with those in normal Krebs' solution. 2. When MgCl2 was added into an organ bath with cumulative way, the magnitude of detrusor contraction decreased dose-dependently. The contractile response was completely abolished by 30 mM MgCl2. 3. Mg ions suppressed the detrusor contraction induced by a high KCl concentration. 4. The effect of Mg ions on the detrusor contraction was enhanced by verapamil but was inhibited by Bay-K8644. 5. Procaine agonized the effect of Mg ions on the detrusor contraction. On the other hand, caffeine did not modify the effect of Mg ions. These results suggest that Mg ions seem to play an inhibitory role on 2 types of Ca-channels existing on the cell membrane and to inhibit release of Ca ions from the intracellular deposit.

Intracellular recording from neurones of the guinea‐pig gall‐bladder.

1. Intracellular recordings were made from neurones of the guinea-pig gall-bladder in vitro. Intracellular injection of horseradish peroxidase revealed a simple structure, consisting of a soma and a single process, but no discernible dendritic arborization. 2. The resting membrane potential was -50.5 +/- 0.4 mV and the input resistance was 80 M omega. 3. Gall-bladder neurones spiked only once at the onset of depolarizing current pulses. Action potentials were blocked by tetrodotoxin, but a Ca2(+)-dependent spike could be elicited in the presence of tetrodotoxin and tetraethylammonium. 4. Action potential after-hyperpolarizations had a duration of 172 +/- 3.7 ms and reversed at a membrane potential of -93 mV; this reversal potential was linearly related to the logarithm of the external potassium concentration. The initial phase of the after-hyperpolarization was inhibited by tetraethylammonium (1-10 mM) and was not affected by 3,4-diaminopyridine. The late phase of the after-hyperpolarization was blocked by apamin (10 nM) or curare (500 microM). Both the early and late phases of the after-hyperpolarization were inhibited when the preparation was perfused with a calcium-free, high-magnesium solution. The calcium-free, high-magnesium solution had no effect on the membrane potential or input resistance of these cells. 5. Fast excitatory synaptic responses and antidromic responses were elicited in gall-bladder neurones by focal stimulation of fibre tracts. High-frequency fibre tract stimulation often resulted in prolonged, calcium-dependent, depolarizations that were associated with a decrease in input resistance. 6. 5-Hydroxytryptamine and substance P caused depolarizations that were associated with a decrease in input resistance. Bethanechol caused hyperpolarizations that were associated with a decrease in input resistance and which were blocked by atropine.

Thyrotropin-releasing hormone causes direct excitation of dorsal vagal and solitary tract neurones in rat brainstem slices

The effect of thyrotropin-releasing hormone (TRH) on neurones in the dorsal motor nucleus of the vagus and the nucleus of the solitary tract was studied using extracellular single-unit recordings from brainstem slices of the rat. About one third of vagal neurones were excited by TRH. The remaining neurones were unaffected. The lowest effective peptide concentration was around 10 nM and a half maximal effect was achieved at about 100 nM. The action of TRH persisted in a low-calcium, high-magnesium solution which blocks synaptic transmission. The biologically inactive compound, TRH-free acid, was without effect. In the nucleus of the solitary tract, one fourth of the neurones were excited by TRH: none were inhibited by this peptide. Part of the vagal TRH-responsive neurones were also excited by oxytocin and some of the solitary tract neurones sensitive to TRH also responded to vasopressin. We conclude that a fraction of neurones located in the dorsal motor nucleus of the vagus and the nucleus of the solitary tract possess functional TRH receptors. TRH may thus act as a neurotransmitter or neuromodulator in the dorsal brainstem and may participate in the regulation of autonomic functions.

Electrophysiological effects of neurotensin on dopaminergic neurones of the ventral tegmental area of the rat in vitro

The effects of neurotensin on the spontaneous firing rate of presumed dopaminergic neurones of the ventral tegmental area of the rat, were studied in a slice preparation of brain by extracellular single-cell recordings. Bath-applied neurotensin excited all cells which were studied ( N = 25). This effect was concentration-dependent; the threshold was 10 −10 M and maximal activation (about 30 spikes/10 sec) was obtained with 10 −6 M. The EC 50 (half-maximal effective concentration) was roughly estimated at 35 nM. The action of neurotensin was mimicked by neurotensin 8–13 ( N = 6), but not neurotensin 1–8 ( N = 6). It persisted in low-calcium, high-magnesium solutions ( N = 5) and therefore probably resulted from a direct activation of neurotensin receptors. The responses to neurotensin were long-lasting (30–60 min after a 10 min 10 −7M infusion) and exhibited little tachyphylaxis. Dose-response curves to the dopaminergic agonist BHT920 showed that, during the infusion of 10 −7M neurotensin, dopaminergic autoreceptors of some neurones were less sensitive than in control conditions. This was not a non-specific effect produced by the excitation, since it was not observed during the infusion of another excitant, N- methyl- d-aspartate (NMDA). These results show that neurotensin potently activates presumed dopaminergic neurones in the ventral tegmental area in vitro; it may also decrease the effectiveness of the autoreceptors of some neurones.

Potassium-induced release of endogenous glutamate and two as yet unidentified substances from the lateral line of Xenopus laevis

The release of endogenous glutamate and other primary amines from the lateral line of Xenopus aevis was studied using an in vitro superfusion technique and high performance liquid chromatography. Potassium stimulation (50 mM KCl) applied to 60 or 120 lateral-line organs dissected from the skin and pooled in a perfusion chamber induced the release of glutamate and aspartate. The release of aspartate was smaller than that of glutamate and more variable. A variable release of two, as yet, unidentified substances was also detected. In low calcium (0.1 mM CaCl 2), high magnesium (10 mM MgCl 2) solution, 50 mM potassium failed to induce an increase in glutamate, aspartate and the two unknowns, suggesting they are released in a transmitter-like manner. The technique presents a new and simple method for studying transmitters in hair-cell systems. Although other interpretations are possible, the results are consistent with the hypothesis that glutamate is a hair-cell transmitter and suggest a potential role for other substances in the transduction process, perhaps as neuromodulators.