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Anesth Analg 2008; 107:1236-1242
© 2008 International Anesthesia Research Society
doi: 10.1213/ane.0b013e31818064f6
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ANESTHETIC PHARMACOLOGY

The Effect of Lidocaine on Cholinergic Neurotransmission in an Identified Reconstructed Synapse

Shin Onizuka, MD, Toshiharu Kasaba, MDC, and Mayumi Takasaki, MD

From the Department of Anesthesiology, Miyazaki Medical College, University of Miyazaki, Kiyotake-Cho, Miyazaki, Japan.

Address correspondence and reprint requests to Dr. Onizuka, Department of Anesthesiology, Miyazaki Medical College, University of Miyazaki, Kiyotake-Cho, Miyazaki 889-1692, Japan. Address e-mail to onizuka{at}shaw.ca.


   Abstract
Top
 Abstract
Introduction
 METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
BACKGROUND: The presynaptic effect of lidocaine on cholinergic synaptic transmission is unclear because of the difficulty in identifying presynaptic neurons and the complexity of the central nervous system in vivo. To clarify the effect of lidocaine on cholinergic synapse, we reconstructed a cultured soma–soma chemical synapse model consisting of two identified visceral dorsal 4 (VD4) and left pedal e-1 (LPeD1) neurons from the snail, Lymnaea stagnalis, in vitro, and used it to determine how lidocaine affects cholinergic synaptic transmission.

METHODS: The response to acetylcholine and excitatory postsynaptic potential (EPSP) amplitude was recorded in the reconstructed chemical synaptic transmission model composed of VD4 and LPeD1 neurons. The currents for acetylcholine measurements were made under voltage-clamp in the presynaptic VD4 and postsynaptic LPeD1 neurons.

RESULTS: Lidocaine inhibited both EPSP and the response for acetylcholine of the postsynaptic neuron. EPSP amplitude was reduced in a voltage-dependent manner in the presynaptic neuron, and lidocaine induced a hyperpolarization shift of the voltage-dependent inactivation curves of EPSP amplitude.

CONCLUSIONS: Lidocaine inhibits cholinergic synaptic transmission with a voltage-dependent inactivation of EPSP amplitude through the membrane potential depolarization of presynaptic neurons.


   Introduction
Top
 Abstract
 Introduction
METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 
Lidocaine is widely used for local anesthesia and postoperative pain relief. Local anesthetics, including lidocaine, induce not only the inhibiting voltage-gated Na+ channels, but also have a wide range of behavioral effects, thus suggesting that they play some role in the central nervous system (CNS).1,2 Among these effects are restlessness, euphoria, muscle twitching, and convulsion, which have been attributed to selective depression or excitation of chemical synapses by local anesthetics. The effect of lidocaine on chemical synaptic transmissions has been discussed. We previously demonstrated that lidocaine induces depression of the inhibitory dopaminergic synaptic transmission, while also inducing excitation in each neuron.3 The question thus remains whether lidocaine affects other chemical synapses, such as acetylcholine (ACh). Local anesthetics have demonstrated a postsynaptic effect on ACh receptors (AChR). Hollmann et al. reported that lidocaine inhibits the cholinergic sensitivity of muscarinic AChR and suggested that the target would be a G protein on the lipid membranes.4 Gentry and Lukas also reported that local anesthetics have a reasonably strong ability to inhibit diverse, human neuronal nicotinic AChR (nAChR) subtypes ({alpha}1, {alpha}3β4, {alpha}4β2, and {alpha}4β4-nAChR), and concluded that local anesthetics act at these nAChR subtypes as noncompetitive functional inhibitors of the nAChR in postsynaptic neurons.5 In contrast, the presynaptic effects of local anesthetics on presynaptic transmissions are still unclear. Lidocaine affects the steady-state inactivation for voltage-dependent sodium channels6 and also voltage-dependent calcium channels.7 These channels affect not only postsynaptic, but also presynaptic transmissions, including transmitter release.8,9 We previously demonstrated that lidocaine induces a depolarization in each pre- and postsynaptic neuron by means of a potassium channel block.3 The depolarization of the presynaptic neuron depresses transmitter release by way of voltage-dependent inactivation of calcium channels and, as a result, postsynaptic neuron excitability cannot be controlled. However, no studies have observed the effect of synaptic transmission on the depolarization of presynaptic neurons by lidocaine. Therefore, the purpose of this study was to determine the effect of lidocaine on neurotransmission in each membrane potential of the presynaptic neuron.

In mammals, it is difficult to investigate the specific neurons, including presynaptic neurons, because of the numerous neurons, small size, long axons, and surrounding glial cells. In contrast, in Lymnaea, these neurons can be individually isolated and the specific synapses can be consistently reconstructed in vitro. This makes it possible to exclude synaptic effects from other neurons, so that the electrophysiological properties can be observed from both the pre- and postsynaptic neurons. This study used a simple synaptic model system which features cholinergic chemical synapse between individually identifiable pre- and postsynaptic neurons from the snail Lymnaea stagnalis.10–12 This soma–soma synapse model provided an excellent opportunity to test both the pre- and postsynaptic effects of lidocaine.


   METHODS
Top
 Abstract
 Introduction
 METHODS
RESULTS
 DISCUSSION
 REFERENCES
 
Animals
All animal experiments were approved by the Animal Care Committee of Miyazaki Medical College. Laboratory-raised stocks of the fresh water snail Lymnaea stagnalis were maintained at room temperature and fed lettuce. For cell culture studies, 3–6 mo old snails with shell lengths of 10–25 mm were selected.

Medium and Saline
Normal Lymnaea saline (NS) was prepared, consisting of 51.3 mM NaCl, 1.7 mM KCl, 4.1 mM CaCl2, 1.5 mM MgCl2, 5.0 mM N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid, and the pH was adjusted to 7.9 with 1 M NaOH. Gentamicin (G-3632, Sigma, USA) was added to a concentration of 150 µg/mL. Defined medium (DM) consisted of serum-free 50% Liebowitz L-15 medium (GIBCO, USA) with added inorganic salts and 20 µg/mL of gentamycin. Inorganic salts in the medium were the same as in normal Lymnaea saline, except that NaCl was decreased to 40 mM and the pH was adjusted to 7.9.13,14

Cell Culture
The left pedal e-1 (LPeD1) and visceral dorsal 4 (VD4) neurons were removed from the snails. After deshelling, the central ganglionic rings were isolated and treated with enzyme for 25 min in DM, using 2 mg/mL trypsin (type III, Sigma). Subsequently, ganglia were treated with 2 mg/mL soybean trypsin inhibitor (Sigma) for 15 min. Neurons were removed by gentle suction with a siliconized, microforge fine-polished pipette with an outside diameter of 1.5 mm (IB-150 F, World Precision Instruments, USA). After removal, the neurons were transferred to poly-l-lysine-coated culture dishes (3001, Falcon, USA) containing 3 mL DM. Neurons were juxtaposed in a soma–soma excitatory configuration as reported previously (Fig. 1A).13,14 After 16–24 h of cell pairing, the evidence for synaptic transmission between the neurons was obtained via simultaneous intracellular recordings as described below (Fig. 1B).


Figure 127
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  		Figure 1. Specific synapses reconstructed between Lymnaea neurons. Identified presynaptic (visceral dorsal 4 [VD4]) and postsynaptic (left pedal dorsal 1 [LPeD1]) neurons were isolated from the visceral and left pedal ganglia, respectively. (A) Neurons were plated in close proximity in defined medium. (B) Simultaneous intracellular recordings were made from both cells. Induced action potentials in VD4 (at arrows) generated 1:1 excitatory postsynaptic potentials (EPSP) in LPeD1. (C) The effects of mecamylamine to the synaptic transmission between the VD4-LPeD1 soma–soma paired cells. (D) The black bar graph shows the EPSP amplitude and the white bar graph shows the amplitude change after an acetylcholine puff. *P < 0.05 in comparison to the control.

 

Intracellular Recording and Perfusion Systems
The membrane potential of both the VD4 and LPeD1 neurons were recorded simultaneously using intracellular recording techniques.15,16 Glass microelectrodes, with an outside diameter of 1.0 mm (GC100-10, CEI, England), were filled with a saturated solution of K2SO4, yielding a tip resistance of 20–60 M{omega}. Neurons were observed under an inverted microscope (TE-300, Nikon, Japan) and impaled by microelectrode. Electrical signals were amplified (IR-283, Neurodata, USA), and recorded on a personal computer through AD and DA converters (Powerlab, ADInstruments, USA).

To investigate the effect of lidocaine on the reconstructed synapse, a presynaptic VD4 neuron was stimulated by 20 mV, 10-ms, single-pulse through a grass-microelectrode for intracellular recording. The electrical synapses by which the membrane potential of the pre-and postsynaptic neurons are synchronized were checked using 20 mV depolarization by a current injection for pre-and postsynaptic neurons, and electrical synapses were thereby excluded in these experiments. Excitatory postsynaptic potentials (EPSP) of LPeD1 to one action potential of the VD4 were measured at each concentration of lidocaine. Exogenous application of ACh (Sigma) was performed (80-ms pulses, 2–3 psi) in some experiments using 1 µM ACh applied directly to the synaptic site via a pneumatic pico-pump (PV800; World Precision Instruments) pressure injector, and the ACh response of LPeD1 was measured. Both the EPSP and ACh response of LPeD1 were measured at –80 mV. The neurons were perfused with NS as a control for 2 min, and then the NS was replaced by lidocaine-containing saline for 5 min. The lidocaine concentrations of 0.01, 0.1, and 1 mM were evaluated. Lidocaine-containing saline had the same osmotic pressure and pH as NS.

Whole Cell Patch-Clamp Recordings
Whole cell voltage-clamp recordings of LPeD1 and VD4 neurons were made using a Multiclamp 700A amplifier (Axon Instruments, Foster City, CA). Patch electrodes (the tip diameter was adjusted to 1 µm, and the resistance was 1–3 M{omega}) were pulled from glass tubing with an outside diameter of 1.5 mm with a filament, (MTW150F-6, World Precision Instruments), on a vertical pipette puller (PA-81, Narishige, Japan). For data acquisition, AD and DA converters (Digidata 1322A, Axon Instruments) were connected to a personal computer. The current was filtered at 1 kHz using a 4-pole Bessel filter and digitized at a sampling frequency of 20 kHz. Data acquisition and analysis were conducted using the p-Clamp 9 software program (Axon Instruments).

To test the cholinergic response for postsynaptic LPeD1 neurons, currents for acetylcholine (IACh) were measured using the whole-cell patch clamp technique. To study IACh, pipettes were filled with a filtered pipette solution containing (pH 7.4) 29 mM KCl, 1/11 Ca2+/BAPTA buffer, and 10 mM N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid, supplemented with 2 mM ATP-Mg and 0.1 mM GTP-Tris. Standard Lymnaea saline (51.3 mM NaCl, mM 1.7 KCl, 1.5 mM MgCl2, and 4.1 mM CaCl2) was used for the bath solution. For IACh measurements, depolarizing step pulses (25 mV/6 s) from –100 to +50 mV were used. All experiments were performed at room temperature (20°C–22°C). An ACh puff was performed (80-ms pulses, 2–3 psi) using 1 µM ACh applied directly to the synaptic site via a pneumatic pico-pump (PV800; World Precision Instruments) with a pressure injector that was linked and controlled on a PC by p-clamp software.17

To clarify whether the cholinergic synaptic transmission was inactivated by the membrane potential of presynaptic neurons, and to elucidate how this was affected by lidocaine, the membrane potential of VD4 was held at –80 mV for 3 min; then, with incremental steps of 40 mV, it was taken to potentials between –160 and +40 mV for 1 s; and, finally, it held at –80 mV for 15 min with the NS perfusion. After this, each concentration of lidocaine, diluted in NS (pH 8), was perfused until the end of this protocol, and the EPSP amplitude was then measured in response to 3 min of 40 mV incremental steps between –160 and +40 mV from a holding potential of –80 mV 10 min after lidocaine perfusion. During this trial, the postsynaptic LPeD1 neuron was held at –80 mV. The EPSP voltage was measured and the inactivation curves were plotted. EPSP voltage was normalized to the maximum in control and plotted as a function of prepulse potential. The plots were fitted to Boltzmann relationships of the form of



Formula 1

where y is the normalized EPSP voltage of the maximum in each group, Vmin is the PSP voltage of baseline, and Vmax is the EPSP voltage of maximum. V is each membrane potential. V50 is the membrane potential at which conductance is halfway between Vmin and Vmax, and K is a slope factor.5 The curve fitting and the analyses were performed using the Igor pro software program (version 5.01, Wave Metrics Inc., Portland, OR).

Statistical Analysis
The results are expressed as the mean ± sd. Changes in the EPSP, InACh, and membrane potential were analyzed by repeated measures one-way analysis of variance, followed by the Scheffé test. Stat view (Abacus, Berkeley, CA) was used for statistical analyses. A value of P < 0.05 was considered statistically significant.


   RESULTS
Top
 Abstract
 Introduction
 METHODS
 RESULTS
DISCUSSION
 REFERENCES
 
Specific Synapse Formation Between Identified Neurons in Cell Culture
A well-established model of identified Lymnaea neurons was used to reconstruct synapses in a soma– soma configuration. Specifically, identified VD4 presynaptic neurons and their postsynaptic partner LPeD1 were isolated from the adult animals and cultured overnight. Within 24 h, these neurons exhibited a chemical synapse (Fig. 1A). Simultaneous intracellular recordings revealed that current injection in VD4 (at arrow, Fig. 1B) generated 1:1 EPSP in a manner similar to those observed in vivo. This synaptic transmission has been shown to be nicotinic acetylcholinergic. Mecamylamine, which is a specific inhibitor for nAChRs, was observed to suppress the nACh response in a concentration-dependent manner (Figs. 1C, D). The ED50 of mecamylamine was 0.76 µM for EPSP and 0.72 µM for1 µM of ACh puff.

Lidocaine Blocked Cholinergic Synaptic Transmission
To test and compare the effects of lidocaine on cholinergic synaptic transmission between VD4 and LPeD1, the soma–soma paired cells were recorded in either the absence or the presence of lidocaine. The synapses were tested electrophysiologically. The action potentials in VD4 under control saline conditions (Fig. 2A) generated 1:1 EPSP in LPeD1, and these excitatory responses were mimicked by exogenous ACh (at arrow), which was pressure-applied directly at the synapse using a fast perfusion system. The perfusion solution was then switched to the saline containing lidocaine, and the synaptic transmission was tested again. Both the synaptic transmission and the cholinergic responses were significantly reduced (EPSP: control, 34.3 ± 9.4 mV; 0.01 mM; 26.3 ± 8.9 mV, 0.1 mM; 11.5 ± 2.5 mV, 1 mM; 0.8 ± 0.7 mV [n = 8 for each case]; cholinergic responses amplitudes: control, 29.8 ± 9.9 mV; 0.01 mM; 23.4 ± 7.7 mV, 0.1 mM; 14.4 ± 7.9 mV, 1 mM; 6.9 ± 5.0 mV [n = 8 for each case]) in a concentration-dependent and reversible manner (Fig. 2B). The cholinergic response for a single LPeD1 neuron was voltage-dependent, and was blocked by lidocaine in a concentration-dependent manner (Figs. 3A, B).


Figure 227
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  		Figure 2. Lidocaine suppressed cholinergic synaptic transmission between the soma–soma paired cells. (A) Intracellular recordings between visceral dorsal 4 (VD4) and left pedal dorsal 1 (LPeD1) revealed a cholinergic synapse, and action potentials in the presynaptic cell generated 1:1 excitatory postsynaptic potentials (EPSP). These excitatory responses were mimicked by exogenously applied acetylcholine (ACh) (at arrow), which generated a compound postsynaptic potential. Both synaptic (VD4) and postsynaptic (ACh) responses were significantly reduced by 1 mM lidocaine. The lidocaine-induced depression was concentration-dependent, and the synaptic transmission returned to its baseline within minutes of washout (Wash) with normal saline (B). The black bar graph shows the EPSP amplitude and the white bar graph shows the amplitude change after an ACh. *P < 0.05 in comparison to the control. #P < 0.05 in comparison to 0.01 mM lidocaine perfusion. {dagger}P < 0.05 in comparison to 0.1 mM lidocaine perfusion.

 

Figure 327
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  		Figure 3. (A) Lidocaine suppressed InACh in the postsynaptic left pedal dorsal 1 (LPeD1) neuron. InACh was measured for a depolarizing step pulse from –100 to +50 mV. Acetylcholine 1 µM was puffed at 1 s in each step pulse. (B) Current-voltage curves of InACh. Closed circles: InACh in control, opened circles: 0.01 mM lidocaine perfusion, opened triangles: 0.1 mM lidocaine perfusion, opened squares: 1 mM lidocaine perfusion, n = 11 in each group. Results are presented as the mean ± sem.

 

A Lidocaine-Induced Hyperpolarizing Shift Voltage-Inactivated the Synaptic Transmission
To test whether the cholinergic synaptic transmission is inactivated by the membrane potential of the presynaptic neuron, and how this is affected by lidocaine, the membrane potential of a VD4 presynaptic neuron was voltage-clamped with incremental steps of 40 mV, between –160 and +40 mV. This demonstrated that cholinergic synaptic transmission is also inactivated by the membrane potential of the presynaptic neuron (Figs. 4A, C) and a lidocaine-induced hyperpolarizing shift in the inactivation curve (V50: control, –40 ± 2 mV [n = 12]; 0.01 mM; –54 ± 3mV [n = 10]; 0.1 mM; –80 ± 4 mV [n = 8]; 1 mM; 104 ± 6 mV [n = 7]) (Figs. 4B, D).


Figure 427
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  		Figure 4. (A) The membrane potential of visceral dorsal 4 (VD4) was held at –80 mV for 3 min then, was taken with incremental steps of 40 mV to potentials between –160 and +40 mV for 1 s in the normal saline perfusion. The excitatory postsynaptic potentials (EPSP) amplitude of left pedal dorsal 1 (LPeD1) was measured at the end of each step pulse. (B): Lidocaine 1 mM was perfused until the end of this protocol, and the EPSP amplitude of LPeD1 was measured at the end of each step pulse that in response to 40 mV incremental steps between –160 and +40 mV from a holding potential of –80 mV 10 min after lidocaine perfusion. (C) In the control, cholinergic synaptic transmission was inactivated after the depolarization of presynaptic VD4 neuron. (D) Inactivation curve of EPSP. Plots were normalized to the maximum InACh in each lidocaine concentration. Lines show the steady-state inactivation curve fitted to Boltzmann relationships.

 


   DISCUSSION
Top
 Abstract
 Introduction
 METHODS
 RESULTS
 DISCUSSION
REFERENCES
 
These results demonstrate that lidocaine suppressed cholinergic synaptic transmission both pre-and postsynaptically in a concentration-dependent manner (Figs. 2 and 3). The effects of lidocaine on synaptic transmission were thought to be primarily postsynaptic effects resembling effects on receptors. There are no reports differentiating the pre-and postsynaptic suppression by lidocaine, because it is difficult to identify the same type of pre- and postsynaptic neurons and the synapse in the CNS of mammals because of their complexity. In contrast, the reconstructed synapse used this study is very simple, and it was possible to reproduce the same types of synapses because, in Lymnaea, most neurons are identified and mapped. In addition, these identified neurons are able to recognize each other and reconstruct a synapse as in vivo. Some synapses are identified by their neurotransmitters. These reconstructed synapses consisted of only two neurons; therefore, individual synaptic transmissions could be observed. In this study, the VD4-LPeD1 synapse was chosen. The neurotransmitter one is ACh. Using this reconstructed synapse demonstrated that the EPSP was inactivated by the voltage-dependent manner of the presynaptic neuron. Lidocaine induced a hyperpolarization shift in the inactivation curves of the EPSP (Fig. 4). Synaptic transmission depends on voltage-dependent sodium and calcium currents, because the transmitter release occurs when the calcium concentration is locally increased to a high level through voltage-dependent calcium channels during an action potential that induces the activation of voltage-dependent sodium channels. Lidocaine induced a hyperpolarization shift in the steady-state inactivation of these currents and this is one reason why lidocaine affects voltage-dependent inactivation of EPSP; however, further study will be required to fully elucidate the mechanism.6,7,18

Lidocaine induces depolarization as shown in previous reports,3,6 and also affects presynaptic neurotransmission. This is because EPSP is decreased in a membrane potential-dependent manner in the presynaptic neuron, as shown in Figure 4. Therefore, lidocaine suppresses presynaptic neurotransmission by depolarization of the presynaptic neurons. Some studies have shown that lidocaine induces depolarization in a neuron; however, the mechanisms are unclear. It is reported that blockade of potassium channels or calcium influx may be the reason for depolarization.19–22 In any case, lidocaine suppresses presynaptic neurotransmission directly by depolarization of presynaptic neurons. However, other mechanisms, such as transportation of neurotransmitters or neurotransmitter release mechanisms, have been shown.23–25 Therefore, more investigation will be required to characterize the effect of lidocaine on presynaptic neurotransmission.

In the trace of Figure 1C, mecamylamine suppressed both EPSP and also the response to ACh puff; therefore, the EPSP and the response to ACh puff of the postsynaptic neuron are considered to demonstrate the ACh response.

In clinical practice, from 40 to 80 mM of lidocaine was used, therefore, a level below 1 mM lidocaine is considered to be a clinically acceptable concentration. However, in the CNS, the concentration of 0.1 mM lidocaine is a toxic dose that may induce convulsions in the rat.26–28 In humans, CNS effects include convulsions, which are also thought to develop progressively with an increase in the plasma concentration up to approximately 18–26 µg/mL (about 0.1 mM).29–31 In this study, the lidocaine concentration was established for a toxic dose and 0.1–1 mM lidocaine showed not only a physiological effect, but also some toxic effects, even though sample neurons demonstrated a good recovery after washout, and we did not observe any permanent demoralization or morphological changes; however, the sample neurons showed both depolarization and excitation. This excitation in each neuron and a failure of the synaptic transmission between the neurons thus seems to be one of the causes of CNS toxicity associated with lidocaine.

In these experiments, LPeD1 and VD4, the cardiac pacemaker neurons of Lymnaea stagnalis, were used to provide a model system. It is possible that there are differences between snail and human brains and sensory neurons. However, the neurotransmitter (ACh) and the receptor (nACh) have been identified in Lymnaea stagnalis. Woodin et al.32 showed, using whole-cell recordings on cultured Lymnaea soma–soma synapses, that, of the excitatory nAChR type(s) expressed by the right pedal dorsal 1 (RPeD1) neuron, EPSP can be blocked by the noncompetitive nAChR blocker Mec, thus suggesting the coexpression of nAChRs by VD4 and RPeD1 recombinant neuron.26 van Nierop et al.33 identified the NACHR subunits of Lymnaea and they also compared them with {alpha}7 subunits of humans and rats. They thus demonstrated that the t11 subunits are in the Lymnaea Ach receptors. In particular, the structure and I–V curve between the Lymnaea nAch A subunit and human and rat {alpha}7 subunits were similar. They also reported that many subtypes of nACh receptors were expressed in each neuron. In LPeD1, the I–V curves for Ach in the LPeD1 that was demonstrated by us were similar in this experiment to the subunit {alpha} of nAch A receptors that was demonstrated by Nierop et al. The expression rate of Lymnaea nAch A subunit in left pedal ganglia was high, whereas the visceral dorsal ganglia was quite low; therefore, I–V curves for Ach in the LPeD1may be close to the I–V curve for Lymnaea nAch A subunit.27 Lymnaea stagnalis has provided an excellent opportunity to study the effects of lidocaine and to be able to generalize the model to humans as well.

In conclusion, these findings demonstrated that lidocaine inhibits cholinergic synaptic transmission with a voltage-dependent inactivation of the EPSP amplitude through the depolarization of the membrane potential in presynaptic neurons.


   Footnotes
 
Accepted for publication May 7, 2008.

Supported, in part, by a Grant-in-Aid (No.12770828) for Scientific Research (A) from The Ministry of Education, Science and Technology of Japan., Marunouchi 2-5-1, Chiyodaku, Tokyo 100-8959, Japan.


   REFERENCES
Top
 Abstract
 Introduction
 METHODS
 RESULTS
 DISCUSSION
 REFERENCES
 

  1. Naguib M, Magboul MM, Samarkandi AH, Attia M. Adverse effects and drug interactions associated with local and regional anaesthesia. Drug Saf 1998;18:221–25[Web of Science][Medline]
  2. Hodgson PS, Liu SS, Batra MS, Gras TW, Pollock JE, Neal JM. Procaine compared with lidocaine for incidence of transient neurological symptoms. Reg Anesth Pain Med 2000;25:218–22[Web of Science][Medline]
  3. Onizuka S, Kasaba T, Hamakawa T, Takasaki M. Lidocaine excites both pre- and postsynaptic neurons of reconstructed respiratory pattern generator in Lymnaea stagnalis. Anesth Analg 2005;100:175–82[Abstract/Free Full Text]
  4. Hollmann MW, Fischer LG, Byford AM, Durieux ME. Local anesthetic inhibition of m1 muscarinic acetylcholine signaling. Anesthesiology 2000;93:497–509[Web of Science][Medline]
  5. Gentry CL, Lukas RJ. Local anesthetics noncompetitively inhibit function of four distinct nicotinic acetylcholine receptor subtypes. J Pharmacol Exp Ther 2001;299:1038–48[Abstract/Free Full Text]
  6. Onizuka S, Kasaba T, Hamakawa T, Ibusuki S, Takasaki M. Lidocaine increases intracellular sodium concentration through voltage-dependent sodium channels in an identified Lymnaea neuron. Anesthesiology 2004;101:110–20[Web of Science][Medline]
  7. Xiong Z, Strichartz GR. Inhibition by local anesthetics of Ca2+ channels in rat anterior pituitary cells. Eur J Pharmacol 1998;363:81–90[Web of Science][Medline]
  8. Platel JC, Boisseau S, Dupuis A, Brocard J, Poupard A, Savasta M, Villaz M, Albrieux M. Na+ channel-mediated Ca2+ entry leads to glutamate secretion in mouse neocortical preplate. Proc Natl Acad Sci USA 2005;102:19174–9[Abstract/Free Full Text]
  9. Rahamimoff R, Butkevich A, Duridanova D, Ahdut R, Harari E, Kachalsky SG. Multitude of ion channels in the regulation of transmitter release. Philos Trans R Soc Lond B Biol Sci 1999;354:281–8[Abstract/Free Full Text]
  10. Hamakawa T, Woodin M, Biorgum M, Painter S, Takasaki M, Lukowiak K, Nagle G, Syed N. Excitatory synaptogenesis between identified Lymnaea neurons requires extrinsic trophic factors and is aintained by receptor tyrosine kinases. J Neurosci 1999;19:9306–12[Abstract/Free Full Text]
  11. Feng ZP, Grigoriev N, Munno D, Lukowiak K, MacVicar BA, Goldberg JI, Syed NI. Development of Ca2+ hotspots between Lymnaea neurons during synaptogenesis. J Physiol 2002;539:53–65[Abstract/Free Full Text]
  12. Woodall AJ, Naruo H, Prince DJ, Feng ZP, Winlow W, Takasaki M, Syed NI. Anesthetic treatment blocks synaptogenesis but not neuronal regeneration of cultured Lymnaea neurons. J Neurophysiol 2003;90:2232–9[Abstract/Free Full Text]
  13. Syed N, Bulloch A, Lukowiak K. In vitro reconstruction of the respiratory central pattern generator of the mollusk Lymnaea. Science 1990;12:282–5
  14. Onizuka S, Takasaki M, Syed NI. Long-term exposure to local but not inhalation anesthetics affects neurite regeneration and synapse formation between identified lymnaea neurons. Anesthesiology 2005;102:353–63[Web of Science][Medline]
  15. Hamakawa T, Feng ZP, Grigoriv N, Inoue T, Takasaki M, Roth S, Lukowiak K, Hasan SU, Syed NI. Sevoflurane induced suppression of inhibitory synaptic transmission between soma-soma paired Lymnaea neurons. J Neurophysiol 1999;82:2812–9[Abstract/Free Full Text]
  16. McKenzie D, Franks NP, Lieb WR. Actions of general anaesthetics on a neuronal nicotinic acetylcholine receptor in isolated identified neurones of Lymnaea stagnalis. Br J Pharmacol 1995;115:275–82[Web of Science][Medline]
  17. Ogata N, Tatebayashi H. Kinetic analysis of two types of Na+ channels in rat dorsal root ganglia. J Physiol 1993;466:9–37[Abstract/Free Full Text]
  18. Kawai N, Niwa A. Neuromuscular transmission without sodium activation of the presynaptic nerve terminal in the lobster. J Physiol 1980;305:73–85[Abstract/Free Full Text]
  19. Kindler H, Paul M, Zou H, Liu C, Winegar D, Gray T, Yost S. Amide local anesthetics potently inhibit the human tandem pore domain background K+ channel TASK-2 (KCNK5). J Pharmacol Exp Ther 2003;306:84–92[Abstract/Free Full Text]
  20. Punke A, Licher T, Pongs O, Friederich P. Inhibition of human TREK-1 channels by bupivacaine. Anesth Analg 2003;96:1665–73[Abstract/Free Full Text]
  21. Punke A, Friederich P. Retigabine stimulates human KCNQ2/ Q3 channels in the presence of bupivacaine. Anesthesiology 2004;101:430–8[Web of Science][Medline]
  22. Gold MS, Reichling DB, Hampl KF, Drasner K, Levine JD. Lidocaine toxicity in primary afferent neurons from the rat. J Pharmacol Exp Ther 1998;285:413–21[Abstract/Free Full Text]
  23. Aasheim G, Fink BR, Middaugh M. Inhibition of rapid axoplasmic transport by procaine hydrochloride. Anesthesiology 1974;41:549–53[Web of Science][Medline]
  24. Lavoie P-A, Khazen T, Filion PR. Mechanisms of the inhibition of fast axonal transport by local anesthetics. Neuropharmacology 1989;28:175–81[Web of Science][Medline]
  25. Miyamoto Y, Muto E, Mashimo T, Iwane AH, Yoshiya I, Yanagida T. Direct inhibition of microtubule-based kinesin motility by local anesthetics. Biophys J 2000;78:940–9[Web of Science][Medline]
  26. Takahashi R, Oda Y, Tanaka K, Morishima HO, Inoue K, Asada A. Epinephrine increases the extracellular lidocaine concentration in the brain: a possible mechanism for increased central nervous system toxicity. Anesthesiology 2006;105:984–9[Web of Science][Medline]
  27. Kim YJ, McFarlane C, Warner DS, Baker MT, Choi WW, Dexter F. The effects of plasma and brain magnesium concentrations on lidocaine-induced seizures in the rat. Anesth Analg 1996;83:1223–8[Abstract]
  28. Yokoyama M, Hirakawa M, Goto H. Effect of vasoconstrictive agents added to lidocaine on intravenous lidocaine-induced convulsions in rats. Anesthesiology 1995;82:574–80[Web of Science][Medline]
  29. Rosenberg PH. Clinical pharmacology and applications of local anesthetics. In: Bowdle TA, Horita A, Kharasch ED, eds. The pharmacologic basis of anesthesiology. New York: Churchill Livingstone; 1994:217–38. Ch 9
  30. Concepcion M. Acute complication and side effects of regional anesthesia. Brown DL, ed. Regional anesthesia and analgesia. Philadelphia: Saunders; 2007:10–30. Ch 24
  31. Mcquillan PM. Foundation of regional anesthesia. Hadzic A, ed. Textbook of regional anesthesia. New York: McGraw Hill; 1996:22–120
  32. Woodin MA, Munno DW, Syed NI. Trophic factor-induced excitatory synaptogenesis involves postsynaptic modulation of nicotinic acetylcholine receptors. J Neurosci 2002;22:505–14[Abstract/Free Full Text]
  33. van Nierop P, Keramidas A, Bertrand S, van Minnen J, Gouwenberg Y, Bertrand D, Smit AB. Identification of molluscan nicotinic acetylcholine receptor (nAChR) subunits involved in formation of cation- and anion-selective nAChRs. J Neurosci 2005;25:10617–26[Abstract/Free Full Text]




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