Electrophysiological study of serotoninergic neuron Cl in the pteropod mollusk Clione

Functional characteristics of cerebral serotoninergic neuron Cl, axons of which terminate at the buccal ganglia [7], were investigated in the pteropod molluskClione. Stimulating neuron Cl induced activation of the feeding rhythm generator located in the buccal ganglia — an effect arising after a long latency and persisting for some tens of seconds once stimulation had ended. Neuron Cl receives feedback from buccal ganglion cells and this brings about periodic modulation in ganglia activity during the generation of feeding rhythm. Activity of neuron Cl is correlated with operation of the locomotor rhythm generator located in the pedal ganglia. The firing rate of Cl neurons increased upon activation of the locomotor generator (whether spontaneous or induced by stimulating certain command neurons). The correlation found between workings of the locomotor generator and activity of Cl neurons is thought to be one of the manifestations of feeding synergy involving simultaneous activation of the locomotor and buccal apparatus.Institute for Research on Information Transmission, Academy of Sciences of the USSR, Moscow. M. V. Lomonosov State University, Moscow. Translated from Neirofiziologiya, Vol. 23, No. 1, pp. 18–25, January–February, 1991.     

Effect of ischaemia on protein synthesis in neuron and glia-enriched fractions from the rabbit spinal cord

The incorporation of 14C-leucine into the post-mitochondrial supernatant and neuron, glia and myelin-enriched fractions isolated from the rabbit spinal cord was studied after ischaemia and subsequent recirculation. In the cell-free system, incorporation decreased to 55% of the control value after 40 min ischaemia, but proteosynthesis returned to the pre-ischaemic value after 3 h recirculation and remained at this level during further recirculation. The incorporation of amino acids into proteins of neurons and neuroglia differed from the cell-free system and from each other. In the enriched neuronal fraction, protein synthesis fall after ischaemia and also during the first hours of recirculation, but during further recirculation it rose to 60% above the control value. In the enriched glial fraction, specific radioactivity of proteins rose abruptly immediately after ischaemia and by the fourth day there was sixfold increase as compared with control values. The results indicate that the ischaemia-induced decrease in protein synthesis is only transient and that a significant increase occurs in the surviving cell populations, especially the neuroglia. The functional changes caused by spinal cord ischaemia are irreversible, however.     

Disturbance of action potential generation in the mollusk giant neuron after preliminary membrane hyperpolarization

A disturbance of action potential generation in the mollusk giant neuron was investigated by intracellular recording. The anodal-breaking effect resulting from cessation of intracellular hyperpolarization was chosen as the factor evoking splitting of the action potential into its separate components. This effect was shown to be accompanied by splitting of the high-amplitude action potential into low-amplitude components. This splitting arises at a certain critical strength of the hyperpolarizing current (the disintegration threshold) and it increases with an increase in the intensity and duration of the hyperpolarizing current. Splitting of the action potential into components is regarded as disturbance of the connections between individual areas of the excitable membrane of the neuron soma. Frequent repetition of hyperpolarization leads to accumulation of the disturbances and intensifies the disintegration of the action potential into its components.     

Calcium ion modulation of short-term plasticity of the cholinoreceptor membrane of the mollusk neuron

Repeated iontophoretic acetylcholine applications to the external surface of the plasma membrane of identified Helix neurones elicit a gradual reversible reduction of cholinoreceptors (ChR) sensitivity. Influence on Ca-conductivity of neuronal membrane modifies the dynamics of lowering of ChR sensitivity. Administration of cadmium ions--blockader of Ca-conductivity, slows down and weakens ChR habituation, while Ca-conductivity activation by raising of extracellular Ca2+ content accelerates and deepens the habituation. It is suggested that chemo-controlled Ca-conductivity takes part in regulation of short-term ChR plasticity.     

Acid habituation in Salmonella enteritidis PT4: impact of inhibition of protein synthesis

Incubation of Salmonella enteritidis PT4 in media at pH values between 3.0–6.0 resulted in a marked and rapid increase in acid resistance, manifested when cells were subsequently challenged at pH 2.5-2.9. Habituation involves two separate systems, one of which is independent of protein synthesis. The relative importance of the two systems is governed by the pH of the challenge medium. Thus in broth at pH 2.7 or above, inhibition of protein synthesis during habituation did not lessen subsequent acid resistance. At pH values of 2.6 and below, the addition of chloramphenicol during habituation was found to reduce subsequent acid resistance although cells were not as sensitive as control cultures grown at pH 7.0.     

Differences in the habituation of the nicotinic and muscarinic cholinoreceptors of Helix neuron PPa4

Pharmacological division of the snail cholinoreceptors population of the identified neurone RPa4 by cholinoblockators of muscarine (atropine, platyphylline) and nicotine (d-tubocurarine) receptors allowed to reveal differences in the dynamics of reversible reduction of sensitivity of these receptors during their habituation to repeated iontophoretic acetylcholine applications. Maximum desensitization of nicotine receptors is weaker, develops slower and is eliminated faster after the end of rhythmic acetylcholine applications. An assumption is made that a more rapid and deeper lowering of muscarine cholinoreceptors sensitivity is due to an increase of intracellular concentration of free calcium at their activation by the agonist.     

Independence of 1,25-dihydroxyvitamin D3-mediated calcium transport from de novo RNA and protein synthesis

The administration of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) to rachitic chicks produces an increase in (a) RNA and protein synthesis, (b) calcium binding protein (CaBP) concentration, and (c) alkaline phosphatase activity in the duodenum. These events occur concomitantly with enhanced calcium transport. We inhibited RNA and protein synthesis in richitic chicks and measured the subsequent response to 1,25(OH)2D3. Actinomycin D, injected prior to and following 1,25(OH)2D3 administration, inhibited intestinal RNA polymerase activity, blocked the rise in serum calcium, reduced the amount of CaBP, and increased alkaline phosphatase activity. Cycloheximide injected in similar fashion, inhibited the 1,25(OH)2D3-mediated increase in intestinal protein synthesis, serum calcium, CaBP, and alkaline phosphatase activity. Neither inhibitor blocked the ability of 1,25(OH)2D3 to stimulate calcium transport as measured in isolated duodenal loops in vivo. The ability of either inhibitor to block 1,25(OH)2D3-mediated calcium transport despite inhibition of CaBP production and alkaline phosphatase activity (by cycloheximide) indicates that de novo RNA and protein synthesis, and in particular CaBP and alkaline phosphatase, are not required for the 1,25(OH)2D3 stimulation of calcium transport.     

The biochemistry of the neuron. Neurosecretory habituation to repetitive depolarizations in PC12 cells

Habituation in response to repetitive depolarization of PC12 cells can be used as a model for memory processes at the molecular level. In response to depolarization by high external potassium, a triphasic elevation in internal calcium levels occurred. Calcium elevation was maximal immediately after addition of the stimulus (phase 1), followed by a 2-min period in which the calcium level decreased (phase 2), leading to a new steady-state level which was higher than in the unstimulated cell (phase 3). In response to repetitive depolarizations, the calcium level in phase 1 was reduced by as much as 43%, and phase 3 was reduced by as much as 40%. By measuring the relationship between calcium elevation and secretion, it was shown that measured reductions in calcium levels were correlated with neurosecretory habituation. One of the components responsible for the reductions in calcium levels was a tetraethylammonium-sensitive potassium channel, and the habituation of this channel was reversed by addition of 4 beta-phorbol 12-myristate 13-acetate.     

Independence of Bacillus subtilis spore outgrowth from DNA synthesis.

The outgrowth of spores of Bacillus subtilis 168 proceeded normally in temperature-sensitive DNA mutants under restrictive conditions and in the absence of DNA synthesis. Two inhibitors of DNA synthesis, nalidoxic acid and 6-(p-hydroxyphenylazo)-uracil, inhibited spore outgrowth under some nutritional conditions; this inhibition of outgrowth however, though not that of DNA synthesis, could be reversed by glucose. The sensitivity of the outgrowing spores to nalidixic acid and 6-(p-hydroxyphenylazo)-uracil inhbition decreased as a function of outgrowth time. The cells became completely resistant to the inhibitors after 90 min. The development of this resistance occurred also in the absence of DNA synthesis. It was concluded that DNA synthesis is not needed for spore outgrowth, and that outgrowing cells and vegetative cells differ in their sensitivity to these inhibitors.     

Polarity and hydrophobicity interactions in protein synthesis process

About 30 years ago, experiments found that there are polarity and hydrophobicity (P and H) correlations and affinity between amino acids and their anticodons. Although it is shown that these experimental findings are important for explaining the origins of the genetic code, the great potential of P and H interactions in investigating other bio-functions have not been fully explored. Here, through raising, discussing and answering seven relevant questions hidden in tRNA aminoacylation, the formation of peptide bonds, and the ending of translations, etc., we show our theoretical findings that the P and H correlations and affinity take vital roles in the protein synthesis process. We found the relationship between the 3' end ACCN sequences of tRNA molecules and the activated amino acids and its biological significance, the rRNAs' consensus sequences 5'NCC...TGG3' or 5'TGG...NCC3' which may perform as functional segments of rRNAs to help triggering the reaction of peptide formation, and common nature of releasing factors that the first amino acid residue of releasing factors ERF, RF1 and RF2 are all Methionine, except a few Alanine, which may be necessary for releasing the translated polypeptide and stopping the translating process. In the terms of P and H correlations and affinity, we provide explanations of why only using the poly (G) as mRNA template cannot get the poly (Gly) in experiments deciphering the genetic code, why Gly often appears in beta turns and why translational bypassing might occur when translating 5'GGAUGA on mRNA. Since amino acids and nucleotides are the subunits, respectively, for composing proteins and nucleic acids, these findings will help in further understanding interactions among the bio-macromolecules. These findings are also helpful for investigating rRNAs, further understanding the protein synthesis process and analysing similar bio-problems, and should be proved useful for experimental biologists.     

Pharmacological characteristics of the acetylcholine and L-glutamate receptors of the subpharyngeal ganglion neuron in the mollusk Zachrysia guanensis

The presence of acetylcholine and L-glutamate receptors sensitive to the cholinoreceptor-specific ligands cytizine and tubocurarine has been demonstrated by means of microapplication of acetylcholine and L-glutamate to the central parietal zone of the subpharyngeal ganglion of the mollusc Zachrysia guanensis. Azidocytizine inhibited the function of acetylcholine receptor but did not affect glutamate receptor. Based on the electrophysiological data and analysis of ligand conformations a model has been designed for a site where L-glutamine is recognized by glutamine receptor.     

Ultrastructural changes in the MP3 neuron of the mollusk Lymnaea stagnalis after cryopreservation of the isolated brain

Investigation of a possibility of long-term storage of frozen (-196 degrees C) viable neurons and nervous tissue is one of the central present day problems. In this study ultrastructural changes in neurons of frozen-thawed snail brain were examined as a function of time. We studied the influence of cryopreservation, cryoprotectant (Me2SO), cooling to 4-6 degrees C, and a prolonged incubation in physiological solution at 4-6 degrees C on dictyosomes of Golgi apparatus, endoplasmic reticulum (ER) cisternae and mitochondria. It has been found that responses of these intracellular structures of cryopreserved neurons to the above influences are similar: dissociation of Golgi dictyosomes, swelling of endoplasmic reticulum cisternae and mitochondrial cristae. Both freezing-thawing and cryoprotectant were seen to cause an increase in the number of lysosomes, liposomes, myelin-like structures, and to form large vacuoles. The structural changes in molluscan neurons caused by cryopreservation with Me2SO (2 M) were reversible.     

Action potentials of the mollusk giant neuron with changes in extracellular concentrations of sodium and calcium ions

The action potential (AP) of the giant neuron of the molluskPlanorbis corneus exhibits an increased sensitivity of the spike overshoot to external sodium concentration in solutions containing a significantly lowered concentration of calcium. These results suggest that during the AP both sodium and calcium ions may act as carriers of the inward-directed current. During repeated responses the role of calcium ions in AP generation increases while that of sodium decreases. A delay in repolarization can occasionally be observed at the beginning of the falling phase of the AP. This delay is considered to be a result of a decrease in efficiency of the repolarizing action of the outward potassium current due to competition from a current entering the cell at the time of the falling phase. Results suggest that the carrier of this inward current is calcium.A. A. Bogomolets' Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 1, No. 1, pp. 109–117, July–August, 1969.     

Cholinoreceptor sensitivity of the mollusk neuron and frog skeletal muscle to acetylcholine and tetramethylammonium analogs

Studies have been made on the sensitivity of cholinoreceptors in identified isolated neuron from the pedal ganglion of the snail Planorbarius corneus and cholinoreceptors of m. rectus abdominis of the frog Rana temporaria to drugs which differ from acetylcholine by the structure either in cationic group, methylene chain, or ester group. Snail cholinoreceptors were found to be less sensitive to changes in the structure of cationic group and more sensitive to the increase in methylene chain from 3 to 4 groups, as compared to frog cholinoreceptors. The sensitivity of both preparations to changes in ester group, as well as to tetramethylammonium was found to be practically the same. Therefore, the sensitivity of neuronal cholinoreceptors in the snail to the effect of acetylcholine and tetramethylammonium analogues does not significantly differ from the sensitivity of cholinoreceptors in the abdominal muscle of the frog.     

The reaction of the N-cholinoreceptors of the isolated neuron of the mollusk Planorbarius corneus to polymethylene-bis-trimethylammonium compounds

Experiments have been made on isolated giant neurones of the mollusc Planorbarius corneus using clamp technique at temperatures 10 and 20 degrees C. The effect of polymethylene-bis-trimethylammonium compounds with 7-18 methylene groups in the molecule (C7...C18) on N-cholinoreceptors with chloride ionic channels was investigated. All these drugs were found to be agonists. Their cholinomimetic activity depends on the number of methylene groups (up to a certain extent) in their structure. This finding stands true also for skeletal muscles of frog and chick, as it had been shown in our earlier experiments. Analysis of membrane current fluctuations showed that the elementary current, the channel opened time, temperature coefficient (Q10) of the neuronal response to application of an agonist and the calculated Q10 of the reaction rate of the agonist with cholinoreceptor did not significantly differ for C8...C18 from the reaction rate of the agonist with cholinoreceptor. As compared with C8, C12...C18 exhibited 30 ... 40 times higher cholinomimetic activity, all other parameters in them being similar. Presumably, this difference is explained by concentrating capacity of C12...C18 at the membrane site because of their higher hydrophobic properties.     

Mollusk neuron habituation following extra- and intracellular administration of an ACTH fragment

By means of intracellular recording of the mollusc neuronal activity a study was made of the influence of ACTH4-7--a fragment of the stress hormone,--on elaboration of habituation to tactile stimuli. ACTH was administered immediately after the elaboration of the series I of habituation (25 to 30 stimuli) by means of either perfusion or extra- or intracellular microionophoresis. This was followed by the II-V habituation series (with 15 min intervals between them). Perfusion and extracellular (not intracellular) ACTH4-7 administration produced an acceleration of habituation in the series II, increase of action potentials (AP) thresholds in the response to the first tactile stimulus of series II and cessation of further increase of the AP thresholds in the course of elaboration. Without ACTH, these phenomena were observed beginning with habituation series IV. The ACTH action while influencing the memory is apparently directed to the external side of the excitable neuronal membrane; ACTH seems to accelerate the consolidation process. Any of the methods of ACTH administration produced also an enhancement of EPSPs and an increase in the number of APs in the responses to stimuli, without lowering the thresholds of APs generation. This testifies toi enhanced chemosensitivity of the membrane and may underlie the influence of ACTH on the level of wakefulness.     

Dopamine modification of the Ca-component of the action potential in the neuron soma of the mollusk L. stagnalis

Dopamine has been found to decrease reversibly the action potential rising phase in identified L. stagnalis neurones. Using voltage-clamp and intracellular dialysis techniques it is shown that dopamine inhibits electroexcitable Ca-current, and changes in Ca-current determine those in the action potential shape. Serotonin and adrenaline influence the Ca-current in a similar way as dopamine does. Inhibition of Ca-conductance is found not to be related to possible effect of these neuromediators on intracellular level of free Ca ions or cyclic AMP.     

Acute effect of exposure of mollusk single neuron to 900-MHz mobile phone radiation

The goal of the present work was to explore the influence of commercially available cell phone irradiation on the single neuron excitability and memory processes. A Transverse Electromagnetic Cell (TEM Cell) was used to expose single neurons of mollusk to the electromagnetic field. Finite-Difference Time-Domain (FDTD) method was used for modeling the TEM Cell and the electromagnetic field interactions with living nerve ganglion and neurons. Neuron electrophysiology was investigated using standard microelectrode technique. The specific absorption rate (SAR) deposited into the single neuron was calculated to be 0.63?W/kg with a temperature increment of 0.1°C. After acute exposure, average firing threshold of the action potentials was not changed. However, the average latent period was significantly decreased. This indicates that together with latent period the threshold and the time of habituation might be altered during exposure. However, these alterations are transient and only latent period remains on the changed level.