Effects of monoamines on heterosynaptic facilitation in giant neurons of Planorbis corneus

The effects of noradrenaline and dopamine on heterosynaptic facilitation (HSF) were investigated during experiments on giant identified neurons from the cerebral ganglion of the freshwater molluskPlanorbis corneus. It was found that catecholamines, while inducing an increase in the amplitude of response obtained by iontophoretic application of acetylcholine, also sharply reduce the amplitude of EPSP occurring as a result of stimulating the cerebral nerve, although acetylcholine also acts as transmitter in this instance. Catecholamines were also found to exert a blocking action on synaptic transmission during the period of HSF. Noradrenaline washout immediately after blockade of HSF was shown to reinstate and promote continuing independent facilitation of synaptic transmission. Duration of this facilitatory after-effect frequently exceeded that of initial HSF. This would imply that if serotonin promotes development of HSF, then the secretion of noradrenaline and dopamine brought about by certain effects act as the mechanism controlling duration and intensity of HSF under naturally-occurring conditions of the mollusk and perhaps even human nervous system activity.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 21, No. 2, pp. 224–232, March–April, 1989.     

Characteristics of heterosynaptic facilitation in giant neurons of the cerebral ganglion of Planorbis corneus

Experiments on giant neurons of the cerebral ganglion of the molluskPlanorbis corneus showed that heterosynaptic facilitation even if evoked by a single uncombined stimulation of the pallial nerve, is more effective than facilitation achieved by a combination of stimulations of nerves directly entering the cerebral ganglion. The intensity of facilitation does not depend on the synaptic efficiency of the heterosynaptic input for the test neuron, but on the intensity of its connection with the other neurons surrounding the giant cell (conjecturally of neurosecretory type). This fact, and also the long latent period of manifestation and achievement of the maximum of facilitation, and its nonspecificity relative to several inputs all suggest that heterosynaptic facilitation is neurosecretory in its origin. Such a mechanism of a sharp increase in the efficiency of synaptic connections in a nerve center may play an important role in the animal's nervous activity as a whole and in the formation of temporary connections in particular, although it does not reflect the specificity of the conditioned reflex.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 12, No. 5, pp. 498–507, September–October, 1980.     

Physicochemical properties of Planorbis corneus erythrocruorin.

The erythrocruorin from the snail Planorbis corneus had a sedimentation coefficient, so/20,w, of 33.5 +/- 0.31 S, and a molecular weight of 1.65 x 10(6) +/- 0.04 x 10(6) by high-speed sedimentation-equilibrium ultracentrifugation. The amino acid composition and absorption spectrum of the protein are reported. A very low number of half-cystine residues was found, corresponding to 0.4 residue per haem group. The haem content was 2.76 +/- 0.22%, corresponding to a protein molecular weight of about 22300. Under both acid and alkaline conditions partial dissociation took place to yield mixtures of products that could not be identified. A subunit corresponding to that containing one haem group was not obtained under any of the dossociating conditions tried. Electron microscopy revealed a ring-shaped molecule about 12.2 +/- 0.5 nm in diameter. The native erythrocruorin bound O2 co-operatively, the intermediate value of h in Hill plots having values between 1.7 and 3.4 depending on the conditions.     

Analysis of heterosynaptic facilitation in identified giant neurons from cerebral ganglion of the pond snail Planorbis corneus.

1. The intracellular mechanism of heterosynaptic facilitation (HSF) formation in identified neurons from the snail Planorbis corneus has been studied. 2. Facilitation of excitatory postsynaptic currents (EPSC) were induced by (a) stimulation of pallial nerve, and (b) addition to extracellular saline of serotonin, NaF, papaverine, theophylline, caffeine or dibutril-cAMP. 3. A depression of EPSC in solutions containing tolbutamide, a cAMP-dependent protein kinase inhibitor was observed. 4. In some cases the similar facilitation or depression of the current induced by acetylcholine application (ACh-current) was found in the same neuron. 5. The effects on ACh-current were distorted in solutions containing caffeine, a well-known activator of calcium ions release from the intracellular depot. 6. According to our findings, we suggest that adenylate cyclase activity of postsynaptic cells could underlie the formation of HSF and it is likely that this activity was modulated by intracellular concentration of calcium ions.     

Planorbis corneus haemoglobin: Circular dichroism and susceptibility to proteases

The purified haemoglobin of Planorbis corneus was subjected to protease digestion and the resulting products characterised by gel filtration and detergent-gel electrophoresis. Small functional subunits of molecular weights approximately 20 000 were obtained corresponding to a single haem group, but multiples of this unit were also always obtained even at high proteolytic enzyme : haemoglobin ratios. This suggested that the subunits of the native molecule (one-tenth containing perhaps ten O₂-binding sites) were made up of single-binding site domains linked by regions of polypeptide chains having different susceptibilities to proteases. The far ultraviolet CD of the native haemoglobin indicated the presence of a high helix content (75–80%) in the protein. The near ultraviolet and visible CD spectra of oxy- deoxy-, and CO-haemoglobin were reported. Planorbis haemoglobin CD was more like that of vertebrate haemoglobins than that of annelid haemoglobins. Nevertheless the Soret CD of Planorbis oxyhaemoglobin had only about half the rotational strength of that of human haemoglobin A, and was halved again upon removal of the ligand. Also in contrast to Lumbricus and human haemoglobins there was only a small decrease in rotational strength in the 260 nm band when Planorbis oxyhaemoglobin was deoxygenated.     

Effect of inhibitors of energy metabolism on potassium accumulation in the neurons of the mollusk Planorbarius corneus

Studies have been made of the effect of inhibitors of energy metabolism on maintenance of Na and K concentrations, as well as on accumulation of Rb (as an analogue of K) in single neurones of the mollusc P. corneus. Concentrations of Na, K and Rb in cells were determined by X-ray spectral microanalysis. Monoiodoacetate, an inhibitor of glycolysis, decreased concentration gradients of K and Na between cells and the external medium, decreasing accumulation of Rb. Cyanide, which is used as an antirespiratory drug, did not significantly affect neither K and Na content, nor Rb accumulation in the neurones of the mollusc, these data being different from those obtained on the nervous tissue of other animals. Monovalent T1 which is capable of inhibiting the respiratory chain and destroying mitochondrial structure, significantly inhibited Rb accumulation in the neurones. The decrease in accumulation of Rb by monoiodoacetate and T1 was less significant as compared with that produced by a specific blocker of Na/K-pump, ouabain.     

Planorbis corneus haemoglobin. Circular dichroism and susceptibility to proteases.

The purified haemoglobin of Planorbis corneus was subjected to protease digestion and the resulting products characterised by gel filtration and detergent-gel electrophoresis. Small functional subunits of molecular weights approximately 20,000 were obtained corresponding to a single haem group, but multiples of this unit were also always obtained even at high proteolytic enzyme: haemoglobin ratios. This suggested that the subunits of the native molecule (one-tenth containing perhaps ten O2-binding sites) were made up of single-binding site domains linked by regions of polypeptide chains having different susceptibilities to proteases. The far ultraviolet CD of the native haemoglobin indicated the presence of a high helix content (75--80%) in the protein. The near ultraviolet and visible CD spectra of oxy- deoxy-, and CO-haemoglobin were reported. Planorbis haemoglobin CD was more like that of vertebrate haemoglobins than that of haemoblobins. Nevertheless the Soret CD of Planorbis oxyhaemoglobin had only about half the rotational strength of that of human haemoglobin A, and was halved again upon removal of the ligand. Also in contrast to Lumbricus and human haemoglobins there was only a small decrease in rotational strength in the 260 nm band when Planorbis oxyhaemoglobin was deoxygenated.     

Factors affecting the cholinomimetic activity of alkyltrimethylammonium compounds in experiments on isolated neurons of the mollusk Planorbarius corneus

It has been shown that the elementary current is independent whereas the duration of channel opening is slightly dependent on the number of methylene groups (from 1 to 9) in the molecule of alkyltrimethylammonium compounds. However, substances with more than 4 methylene groups exhibit lower cholinomimetic activity (i.e. the ability to increase the membrane current) and higher values of Q10 for the reaction with cholinoreceptor. It is suggested that lower activity of these compounds is due to a low rate of formation of a complex with cholinoreceptor because of the higher potential energy barrier.     

Functional organization of afferent inputs of neurons of the cat somatic cortex

Responses of cortical neurons in the posterior sigmoid gyrus to stimulation of two points of the ventro-posterolateral nucleus were investigated in cats immobilized with D-tubocurarine. Some neurons responding to stimulation of one point of this nucleus with a latent period of 2.5–4 msec, were activated by stimulation of the other point after 10 msec or longer. Conditioning stimulation of one point facilitated or inhibited the response to test stimulation of the other point. The facilitatory effect was usually exhibited if the response latency exceeded 5 msec. It is concluded that a cortical input neuron for some afferent fibers is activated by other similar afferent fibers only after intracortical relay. The system of "input" cortical neurons is thus not only a structural, but also to some extent a functional, concept; under certain conditions an incoming afferent volley activates them only polysynaptically.     

Actions of acetylcholine on the salivary gland cells of the pond snail, Planorbis corneus

Intracellular recordings have been made from salivary gland cells of the pond snail Planorbis corneus. Gland cells produced a dose-dependent biphasic response to the bath application of acetylcholine (ACh), an initial depolarization being followed by a hyperpolarization. Nicotine and the nicotinic agonist tetramethylammonium had an excitatory action on the gland cells. The muscarinic agonists acetyl-beta-methyl choline and arecoline were also stimulants, but muscarine, bethanechol and pilocarpine produced no response from gland cells at 10(-3) M. A number of cholinergic antagonists, including atropine, hexamethonium and curare, effectively blocked the response to ACh. The depolarizing phase of the ACh response resulted from an increased membrane permeability to Na+ ions, though the participation of other ionic species cannot be ruled out. The hyperpolarizing phase of the ACh response was produced by the activity of an electrogenic Na+/K+ pump.     

Responses of afferent and efferent neurons to auditory inputs in the vestibular nerve of the frog

Summary In the frog, the spontaneous discharges of afferent fibres from the horizontal semicircular canal (HC) and of efferent vestibular units were recorded by means of glass micropipettes filled with 2 mol/l NaCl as well as during acoustic stimulation; pure tones 300–2,000 Hz and clicks 150/s, 80–100 dB re 10^–5 N/m² were used. The activity of 56% of the efferent fibres recorded was increased by such stimulations while the discharge of the others was not modified. In intact preparations the activity of 34.4% of the afferent fibres recorded was either increased or decreased by sound stimulation depending on the unit; the discharge of the others (65.6%) was not modified (Fig. 3). Section of both saccular nerves did not change the percentage of the units modulated by sound showing that the saccules have probably no effect on this modulation (Fig. 4). In preparations where the contralateral auditory papillae were eliminated, 21.1% of the afferent units were facilitated and no unit was inhibited (Fig. 5), while in preparations where the ipsilateral auditory organs were eliminated 21.1% of the afferent units were inhibited and no unit was facilitated (Fig. 6). Therefore, in intact preparations one can assume that decrease and increase of the HC afferent fibre discharges were due to stimulation of the contralateral and the ipsilateral auditory organs, respectively. Such a modulation of canal afferent discharges being mediated by efferent vestibular fibres, it can be postulated that the efferent vestibular system has a double influence upon the hair cells of the vestibular epithelium: one inhibitory and the other facilitatory. Such a double effect is discussed.Abbreviations EVS efferent vestibular system - HC horizontal semicircular canal