Storage and erasure of behavioural experiences at the single neuron level.

Although predictions from the past about the future have been of major interest to current neuroscience, how past and present behavioral experience interacts at the level of a single neuron remains largely unknown. Using the pond snail Lymnaea stagnalis we found that recent experience of terrestrial locomotion (exercise) results in a long-term increase in the firing rate of serotonergic pedal (PeA) neurons. Isolation from the CNS preserved the "memory" about previous motor activity in the neurons even after the animals rested for two hours in deep water after the exercise. In contrast, in the CNS, no difference in the firing rate between the control and "exercise-rested" (ER) neurons was seen. ER snails, when placed again on a surface to exercise, nevertheless showed faster locomotor arousal. The difference in the firing rate between the control and ER isolated neurons disappeared when the neurons were placed in the microenvironment of their home ganglia. It is likely that an increased content of dopamine in the CNS masks an increased excitation of PeA neurons after rest: the dopamine receptor antagonist sulpiride produced sustained excitation in PeA neurons from ER snails but not in the control. Therefore, our data suggest the involvement of two mechanisms in the interplay of past and present experiences at the cellular level: intrinsic neuronal changes in the biophysical properties of the cell membrane and extrinsic modulatory environment of the ganglia.

Light-dark decision making in snails: Do preceding light conditions matter?

We have recently demonstrated that preceding motor activity can facilitate decision-making not only in humans and mammals but also in molluscs.5 In the behavioral paradigm used, snails Lymnaea stagnalis were removed from their natural environment (water) and placed in a dry, asymmetrically lit arena from which they had to decide which way to go in order to reach an aquatic environment. One possible explanation of the observed effects of preceding motor activity was that it could affect the snail's memory of light conditions that corresponded to its previous aquatic habitat. Here we report experimental results discarding this hypothesis. We suggest that preceding intense locomotion is likely to facilitate decision-making by increasing the level of confidence.

Serotonin precursor (5-hydroxytryptophan) causes substantial changes in the fighting behavior of male crickets, Gryllus bimaculatus.

This study demonstrates that injection of the serotonin precursor 5-HTP causes substantial changes in the behavioral state, fighting behavior and ability to establish winner-loser relationships in male crickets (Gryllus bimaculatus). The characteristic features of 5-HTP-treated crickets include an elevated posture, enhanced general activity, longer duration of fighting, enhanced rival singing and a decreased ability to produce a clear fight loser. In addition, 5-HTP-treated males showed a slightly delayed latency to spread their mandibles, a decreased number of attacks and an equal potential to win in comparison to controls (physiological solution-treated males). The obtained results imply a significant role for serotonin in the regulation of social status-related behaviors in G. bimaculatus. Specifically, these data indicate that a decrease in serotonergic activity may be functionally important for the control of loser behavior and that some behavioral features of dominant male crickets are likely to be connected with the activation of the serotonergic system.

[The central pattern generators].

The central pattern generator (CPG) is defined as a set of neurons involved in joint production of patterned motor output. The roundtable discussion on the CPG was a part of the 5th All-Russian Conference on Animal Behavior (Moscow, Nov. 21, 2012). The discussion centred on three core themes: 1) the mechanisms of the organization and reconfiguration of pattern generating neuronal ensembles, 2) extrapolations that extend the CPG concept beyond the motor systems, and 3) evolutionary and developmental aspects of CPG.

Electrical activity of no-producing neuron depends on NO level.

NO-producing neuron exhibited an excitatory response to the decrease in NO concentration, which was induced by NO synthase inhibitor N-nitro-L-arginine or specific NO acceptor 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide. Addition of NO donors to the medium inhibits neuronal activity. The excitatory effects of N-nitro-L-arginine and 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide were preserved, while the inhibitory action of NO donors significantly decreased after isolation of the neuron. These findings indicate that NO regulates activity of these neurons by the negative feedback mechanism. This regulation includes the following complementary mechanisms: (1) endogenous mechanism of cell self-activation in response to the decrease in NO concentration; and (2) exogenous mechanism of cell-mediated inhibition in response to NO excess.

Social aggressiveness of female and subordinate male crickets is released by opiate receptor antagonist.

1. In the cricket Gryllus bimaculatus, effects of opiate receptor antagonist naloxone, 9 or 30 microg per animal, on aggressive behavior were investigated. 2. Naloxone had no significant impact on aggression of isolated and dominant males. In contrast, the drug caused a dramatic release of social aggression in female and subordinate male crickets. 3. The results suggest that activity of the opioid system contributes to suppress aggression in subordinate males, as well as in females, during social contacts.

Effects of naloxone on c-jun/AP-1 in met-enkephalin-and FMRFamide-immunreactive neurons of a gastropod snail.

1. Opioid- and FMRFamide (FMRFa)-ergic systems are believed to play antagonistic behavioral roles in both higher and lower animals. In our previous experiments on a snail, behavioral choice has been demonstrated to be dependent on a balance between FMRFa and enkephalins [7]. Here, we examined if the disturbance of the balance causes changes in the activity of both systems. Opiate receptor blocker naloxone was applied and its effect on c-jun expression of met-enkephalin (MEnk)- and FMRFa-ergic neurons was examined immunocytochemically in terrestrial gastropod snail Cepaea nemoralis. 2. In control, untreated snails, central neurons with c-jun/AP-1-like-immunoreactivity were found to occur. These included MEnk-, FMRFa- and 5HT-immunoreactive (-ir) neurons, as was revealed by double-labelling. 3. After treatment with naloxone for 4 h, the following changes were observed: (i) increase in the number of MEnk-ir neurons; increase in the number of neurons showing c-jun/AP-1 and MEnk double-labeling; (ii) disappearance of c-jun/AP-1-immunoreactivity from some FMRFa-ir neurons. 4. It is suggested that immediate early genes are involved in the mechanisms responsible for the reciprocal regulation of the opioid and antiopioid neuropeptide systems.

Effects of serotonergic and opioidergic drugs on escape behaviors and social status of male crickets.

We examined the effects of selective serotonin depletion and opioid ligands on social rank and related escape behavior of the cricket Gryllus bimaculatus. Establishment of social rank in a pair of males affected their escape reactions. Losers showed a lower and dominants a higher percentage of jumps in response to tactile cercal stimulation than before a fight. The serotonin-depleting drug alpha-methyltryptophan (AMTP) caused an activation of the escape reactivity in socially naive crickets. AMTP-treated animals also showed a lower ability to become dominants. With an initial 51.6+/- 3.6% of wins in the AMTP group, the percentage decreased to 26+/-1.6% on day 5 after injection. The opiate receptor antagonist naloxone affected fight and escape similarly as AMTP. In contrast to naloxone, the opioid agonist [d-Ala(2), N-Me-Phe(4), Gly(5)-ol]-enkephalin decreased escape responsiveness to cercal stimulation in naive and subordinate crickets. We suggest that serotonergic and opioid systems are involved in the dominance induced depression of escape behavior.

Anatomical basis for interactions of enkephalins with other transmitters in the CNS of a snail.

Immunocytochemical techniques for double staining were employed to investigate the morphological basis for interactions between enkephalins and other neuroactive compounds in the behavior of the gastropod mollusc Cepaea nemoralis. Coexistence of each of the two enkephalins with FMRFamide, serotonin or GABA-like immunoreactivity was found in certain neurons in cerebral, parietal, and pedal ganglia. Tyrosine hydroxylase-immunoreactive neurons were occasionally seen in close apposition to, but never colocalized with, the enkephalins. A comparison between these anatomical observations and previous behavioral studies suggests that in gastropod molluscs cotransmission of enkephalins with classical transmitters may, at least partly, reflect synergism of these substances in the control of definite behavioral programs.

Complex avoidance behaviour and its neurochemical regulation in the land snail Cepaea nemoralis.

1. In hot plate experiments, the pulmonate land snail Cepaea nemoralis displays a biphasic passive/active avoidance behaviour composed of retraction and subsequent searching mediated by antagonistic muscular systems. The switch, between the behaviours, is under neuronal control. 2. Leu- and met-enkephalin, as well as FMRFamide-antiserum, attenuated the retraction response and potentiated the searching behaviour. Opposite effects were achieved by injection of antisera to the enkephalins. 3. Both retraction and searching behaviours were potentiated by 5-HT. Methysergide antagonized the effects of the enkephalins on the searching behaviour. 4. We conclude that endogenous opioids act antagonistic to FMRFamide in the neuronally controlled switch between passive and active avoidance behaviour.

Mode of action of antipsychotic drugs: lessons from simpler models.

The monoaminergic system of the pond snail Lymnaea stagnalis changed markedly following prolonged exposure to chlorpromazine (CPZ). HPLC-ED indicated that levels of serotonin (5-HT), but not those of dopamine, were significantly decreased (e.g., to less than 40% after 30 days of treatment with 1 microM CPZ). Glyoxylate-induced fluorescence was depressed to undetectable levels in selected subpopulations of 5-HT neurons. Performance of 5-HT-dependent motor behaviors was impaired, and a considerably decreased firing activity was observed in affected 5-HT neurons. The present results, in accord with past ones with haloperidol, suggest that a previously overlooked mechanism of monoamine down-regulation may contribute to affects of antipsychotic drugs.