Completely isolated neurons in the mollusc, Lymnaea stagnalis. A new objective for nerve cell biology investigation

doi:10.1016/0300-9629(74)90544-1

Comparative Biochemistry and Physiology Part A: Physiology

Volume 49, Issue 1, 1 September 1974, Pages 89-92, IN3-IN4, 93-100

Comparative Biochemi...

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Abstract

1.
1. The complete isolation of neurons from the mollusc, Lymnaea stagnalis, has been achieved by enzyme treatment and mechanical isolation.
2.
2. Modifications of the method allow preparation of neurons both with and without axons and with or without glial cells.
3.
3. The isolated neurons maintained their capability to synthesize RNA. Their electrical characteristics were normal.
4.
4. In these experiments the isolated neurons maintained their characteristics in saline for 8–10 hr and under tissue culture conditions for 14 days.

References (7)

C.F. Chen et al.
Pacemaker properties of completely isolated neurons in Aplysia californica
Nature, Lond.
(1971)
T.L. Dyakonova et al.
Peculiarities of structural and functional organization and metabolic activity of the Lymnaea stagnalis neurons. A deposit
Viniti
(1969)
V.I. Geletyuk et al.
Electrical properties of neurons of the mollusc Lymnaea stagnalis under conditions of tissue culture
Tsitologiya
(1972)

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Maintenance of helix pomatia neurons in simplified cell culture system
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Helix pomatia neurons were successfully maintained in primary cell culture using the plant lectin concanavalin A (con A) as the substrate. No additional factors such as hemolymph, conditioning medium or intact ganglia were used. Most of the isolated neurons including ones with large soma diameter (> 200 μm) adhered to the surface and nearly 100% of the adhered neurons sprouted and grew extensively branching neurites. By making connections with each other, the neurites belonging to several neurons often form extensive networks. Spontaneous bioelectrical activity was recorded in about 10% of neurons. In all other cells tested, action potentials similar to ones recorded in situ could be evoked by depolarization up to the 15th day after plating the neurons.
Effects of territrem-B on cholinergic responses of snail neuron
1993, Neuroscience Letters
Effects of territrem-B (TRB), a mycotoxin isolated from a rice culture of Aspergillus terreus, on the central neuron of the snail Achatina fulica were studied electrophysiologically. Territrem-B potentiated the acetylcholine (ACh) induced current of the neuron, while it had no effect on GABA or l-glutamate elicited currents. TRB and neostigmine increased the peak amplitude of the response elicited by the first perfusion of ACh and depressed the increase in current produced by a second perfusion. TRB and neostigmine showed different dose-dependent effects on ACh responses. The results suggested that TRB is a good tool for studying the physiological role of AChE in central neurons.
Effect of deltamethrin on acetylcholine-operated ionic channels in identified Helix pomatia L. neurons
1991, Pesticide Biochemistry and Physiology
The main action of pyrethroids is to slow the gating kinetics of the Na channel of the axon and some. However, it was also found that pyrethroids may affect the neurotransmission as well, acting postsynaptically. It can be proposed, therefore, that pyrethroids may have their target at the synaptic site in addition to the axonal or somal Na channels. The aim of the present study was to examine the effect of deltamethrin on acetylcholine (ACh) receptors of snail neurons using a two-microelectrode voltage clamp. Ionophoretic application of ACh on the soma of identified Helix neurons produced inward current of which amplitude was dependent on the holding potential. Deltamethrin (1–100 μM) depressed the ACh-induced current in a concentration-dependent manner. The I–V curves, both in control and in deltamethrin-containing saline, were linear in the potential range of −100 and −25 mV. The reversal potential was unchanged. The data suggest a monomolecular binding of deltamethrin to the ACh-activated channel with a K_D of 50 μM. Deltamethrin increased the time of recovery from ACh desensitization. It can be concluded, therefore, that deltamethrin may contribute to its toxic effect by affecting the function of the ACh-receptors, although this effect could be secondary.
Electrophysiological detection of acetylcholinesterase activity using concentration clamp on physically isolated Aplysia neurons
1988, Neuroscience Research
In this study, we have used electrophysiological techniques to evaluate the acetylcholinesterase (AChE) activity on single neurons physically isolated from the pedal ganglia of Aplysia californica and kurodai. Acetylcholine (ACh) was applied to cells having Na-dependent response using a concentration clamp technique which allows a step-like complete change of external medium around the cell (diameter more than 120 μm) within 30 msec. When the neuron was exposed to ACh (50 μM) initially by rapid and brief (400–800 msec) flow and continuously after stopping the perfusion, the induced current rose to a peak and then decayed in the presence of ACh. However, the current increased again when the rapid flow of the same ACh solution around the cell was resumed. The increase in the current by the resumption of the perfusion was not seen when carbachol was substituted for ACh or when extraordinarily high concentration (10 mM) of ACh was used. Furthermore, this increase in the current was blocked by the antiAChE agent, edrophonium, in a dose-dependent manner. These results suggest that acetylcholinesterase (AChE) causes a local depletion of ACh at the membrane surface where the rate of hydrolysis of ACh exceeds the rate of ACh diffusion from bulk solution and the increase in the current by the resumption of the perfusion resulted from the restoration of ACh concentration at the membrane surface. This electrophysiological indication of AChE activity may be a useful tool for the study of AChE by other than biochemical means.
On the status of the study of invertebrate neurons in tissue culture-Phyla mollusca and annelida
1987, Comparative Biochemistry and Physiology -- Part A: Physiology
- 1.
  1. The utilization of tissue culture in neurobiological studies is discussed for all phyla phylogenetically preceding Phylum Arthropoda. Only two phyla, Mollusca and Annelida, are represented in such studies.
- 2.
  2. The members of Phylum Mollusca which have been so investigated are Aplysia, Helisoma and Lymnaea.
- 3.
  3. The mollusc Aplysia has been used to investigate several processes, including neurosecretion, synaptic transmission and synaptogenesis.
- 4.
  4. Helisoma was employed to study factors regulating neurite growth and the specificity of synapse formation; mechanisms of neurite growth were investigated in the snail Lymnaea.
- 5.
  5. The only member of Phylum Annelida involved in appropriate studies has been the leech Hirudo. This organism was used to investigate axonal regeneration and synaptic mechanisms.
Neurotransmitter-induced modulation of neuronal Ca current is not mediated by intracellular Ca<sup>2+</sup> or cAMP
1985, Brain Research
Dopamine (1 μM) inhibits the Ca component of action potential and corresponding electroexcitable Ca current (I_Ca) in isolated snail neurons. Adrenaline and serotonin also reduce I_Ca. The inhibition is not related to changes in intracellular Ca²⁺ or cAMP concentrations: internal application of 10 mM ethyleneglycoltetraacetic acid or 10 μM cAMP in the mixture with 1 mM Mg-ATP and 2 mM theophylline does not influence the action of neurotransmitters on I_Ca.

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Comparative Biochemistry and Physiology Part A: Physiology

Abstract

References (7)

Pacemaker properties of completely isolated neurons in Aplysia californica

Nature, Lond.

Peculiarities of structural and functional organization and metabolic activity of the Lymnaea stagnalis neurons. A deposit

Viniti

Electrical properties of neurons of the mollusc Lymnaea stagnalis under conditions of tissue culture

Tsitologiya

Cited by (33)

Maintenance of helix pomatia neurons in simplified cell culture system

Effects of territrem-B on cholinergic responses of snail neuron

Effect of deltamethrin on acetylcholine-operated ionic channels in identified Helix pomatia L. neurons

Electrophysiological detection of acetylcholinesterase activity using concentration clamp on physically isolated Aplysia neurons

On the status of the study of invertebrate neurons in tissue culture-Phyla mollusca and annelida

Neurotransmitter-induced modulation of neuronal Ca current is not mediated by intracellular Ca<sup>2+</sup> or cAMP