The monosynaptic connection: identified synapses in the CNS of the edible snail

By electrophysiological and microanatomical methods of analysis of snail CNS small neurones it was shown that a number of neurones form a monosynaptic connection (MSC) with the gigantic polyfunctional neurone LPa3. By using cobalt and nickel staining, the structure of MSC cells LPa7--LPa3 was studied. Six identified synapses in two LPa3 processes zones were found. Physiological analysis showed that the revealed MSC was plastic. The described MSC with identified synapses is convenient for studying synaptic transmission mechanisms.     

Functional characteristics of an identified pair of neurones in the CNS of the pond snail (Lymnaea stagnalis L.).

The properties of 2 giant electrically coupled neurones (A10 and P1) identified in the visceral and right parietal ganglia of Lymnaea stagnalis were examined. The active and passive electrical parameters of the neurones, as well as the junction between them were measured. The main peripheral and interneuronal connections of the neurones were demonstrated using both electrophysiological and morphological methods. It is shown that the coupled cells are not neurones of the same function, but they are asymmetrical ones. This finding is supported by the following results: (1) the axonal pathways of neurones A10 and P1 are different; (2) there are significant differences in their afferent and efferent connections; (3) though the electrical junction between them is bidirectional, the junctional electrical characteristics prefer P1-A10 transmission. According to the electron microscopic results both neurones are possible neurosecretory cells. The differences demonstrated between the 2 giant neurones may have significance concerning their role in a special neuronal network.     

Reflection of the plastic properties of very simple neuronal assemblies in the statistical characteristics of the spike activity of their elements. Monosynaptically connected neurons

Changes of cross-correlation histograms (CCH) of impulse trains and of mean interspike intervals (ISI) of monosynaptically connected neurones under changes of interneuronal connection efficiency, neuronal excitability and action on these neurones of independent irregular afferent synaptic inflows, were studied by methods of mathematical and biomathematical modelling of synaptic neuronal interaction (computer-controlled experiment on mollusc neurones). It was shown that statistical of increase of efficiency of monosynaptic excitatory or inhibitory interneuronal connection (amplitude enhancement of corresponding postsynaptic potential) is an increase of the main peak or trough of normalized CCH of impulse trains accompanied by a decrease of mean ISIs of both neurones.     

Cellular reactions to elaboration of a backward conditioned connection in Helix lucorum

After 10-15 food stimuli paired with electrical shock in semi-intact snail preparation, responses to strong tactile stimuli identified feeding behaviour neurones were studied. Inhibition evoked by tactile stimulation in these cells before learning procedure disappeared and in some cases noxious stimulus evoked synaptic activation corresponding to feeding reactions in the intact animal. Changes in second-order sensory neurones pre-synaptic to the command neurones of avoidance behaviour are suggested to be the mechanism of forward conditioned connection as well as the mechanism of backward conditioned connection.     

Plant-odour-specific receptor neurones on the antennae of female and male Spodoptera littoralis

Abstract. Receptor neurones with high selectivity and sensitivity to plant odours were found within short sensilla trichodea on the antenna of both female and male Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae) by using single-sensillum recording techniques. In 112 sensilla from females and forty-one from males, twenty-four different receptor neurone types were characterized according to their specificity. Altogether, twenty-six plant and three sex pheromone compounds were tested. Receptor neurones responding with high specificity to flower odours, green leaf volatiles, oviposition deterrents and other general host plant odours were identified. In twenty-one receptor neurone types, responses were elicited by one or several plant compounds, and in three types responses were elicited by sex pheromone compounds. The majority of the receptor neurones responded to only one or two of the tested compounds. In general, only one of the two receptor neurones in a sensillum responded to any of the compounds tested. An exception was a receptor neurone responding to plant odours (green leaf volatiles) and another receptor neurone responding to a sex pheromone compound ([Z]-7-dodecenyl (acetate), which occurred in the same sensillum. The majority of the receptor neurones displayed a high sensitivity to plant odours. No morphological difference was identified the different sensillum types.     

Sensory responses and axonal morphology of two different types of cerebral neurones in Helix pomatia L

1. The mechano- and chemosensory responses of two different types of cerebral neurones responding to food stimuli were studied by microelectrophysiological techniques. This was correlated with axonal morphology investigated by intracellular Co-lysine labelling. 2. The C4 neurone responded to food stimuli applied to the lip receptors by phasic (less than 30 sec duration), while the C14 neurone by an initial phasic and a subsequent tonic longer than 10 min duration) activity change. 3. The different sensory responses of the two neurones can partly be explained by the morphological differences found between them.     

Phenotypic differences in the GnRH neuronal system of deer mice Peromyscus maniculatus under a natural short photoperiod

The neural mechanisms by which short photoperiod induces gonadal regression among seasonally breeding mammals are not well understood. One hypothesis suggests that the proximate cause of seasonal gonadal regression is a photoperiod-induced modification in GnRH secretion. This hypothesis is indirectly supported by our recent findings using immunocytochemistry which identified specific photoperiod-induced adjustments in the number and morphology of GnRH containing neurones between reproductively competent and reproductively regressed laboratory housed male deer mice. Herein, we report that the GnRH neuronal system is similarly affected in reproductively responsive and nonresponsive wild male deer mice Peromyscus maniculatus exposed to a natural short photoperiod. The distribution of immunoreactive (IR)-GnRH neurones was nearly identical in field caught animals and those housed under artificial photoperiod in the laboratory. Compared with reproductively nonresponsive males, reproductively responsive mice from the field population possessed a greater total number of IR-GnRH neurones, a greater number of IR-GnRH neurones within the lateral hypothalamus, and a greater proportion of bipolar IR-GnRH neurones. Each of these distributional and morphological characters was consistent with our findings in laboratory housed male deer mice exposed to an artificial short photoperiod. Taken together, these data underscore the validity of using an artificial photoperiod to evaluate seasonal adjustments in reproductive function in the laboratory.     

Significance of nitric oxide function in development of cardiac hypertrophy under condition of experimental renal hypertension

The development of cardiac hypertrophy was studied under condition of experimental renal hypertension on the rat. The number of cardiac nitric oxide synthase (NOS)-positive neurones increased simultaneously with the increase in NOS-activity in these neurones. A connection was found between the development of cardiac hypertrophy and the activity of NOS in cardiomiocytes. The involvement of NO in the development cardiac hypertrophy as auto- and paracrine regulator is supposed.     

On coupling and decoupling of spinal interneuronal networks

This review addresses the question of interrelations between spinal interneuronal networks. On the basis of electrophysiological, pharmacological, morphological and immunohistochemical analysis of interneurones mediating various reflex actions from muscle receptors and of reticulospinal neurones a considerable degree of interweaving between networks of these neurones has been established. The coupling has been found to occur at the level of several sites of these networks but the review focuses on two of these sites. The first is between dorsal horn interneurones with group II input and their target ipsilaterally and contralaterally projecting intermediate zone and commissural interneurones. The second is between commissural interneurones with input from reticulospinal neurones and their target interneurones. Several ways of both strengthening and weakening of coupling between various interneuronal networks are also briefly reviewed.     

Green leaf volatile-detecting olfactory receptor neurones display very high sensitivity and specificity in a scarab beetle

In the Japanese scarab beetle, Phyllopertha diversa, olfactory receptor neurones specific for the detection of so-called general green leaf volatiles (GLV) display a high specificity and sensitivity. Three main types of green-leaf-volatile-detecting receptor neurones specific to (Z)-3-hexenyl acetate, (E)-2-hexenal and (Z)-3-hexenol, respectively, were identified. Each type responded at a very low stimulus concentration to the key stimulus, and required at least a thousand-fold increase in concentration to respond to any of the other GLVs tested. Flower-odour-and pheromone-detecting receptor neurones were also identified. Olfactory sensilla housing plant-odour- or pheromone-detecting receptor neurones displayed clear morphological differences, and were also separated into different antennal regions.     

Effects of synthetic biologically active peptides on giant neurones identified in the left buccal ganglion of an African giant snail (Achatina fulica Férussac)

1. The effects of synthetic biologically active peptides, including Met-enkephalin, substance P, oxytocin, Arg-vasopressin, proctolin and FMRFamide, on the following four buccal neurones were examined: d-LBAN (dorsal-left buccal anterior neurone), d-LBMN (dorsal-left buccal medial neurone), d-LBCN (dorsal-left buccal central neurone) and d-LBPN (dorsal-left buccal posterior neurone). These peptides were examined at 10(-4) M. 2. Oxytocin excited d-LBAN and slightly excited d-LBCN, while this inhibited d-LBMN. Arg-vasopressin excited slightly d-LBAN and d-LBCN, but this had some times no effect. FMRFamide inhibited d-LBAN, and slightly inhibited d-LBCN. 3. No direct synaptic connection from the two ventral cerebral giant neurones, v-LCDN and v-RCDN, to the four buccal giant neurones was found, though the two cerebral neurones innervate the cerebro-buccal connectives.     

Golgi-Cox studies on the central nervous system of a gastropod mollusc

Summary Complete neurones were impregnated in the brain of the pulmonate gastropod pond snail, Lymnaea stagnalis L. using the Golgi-Cox method. Mapping of small to medium sized neurones identified in living preparations by the position of the perikarya was possible. Simple monopolar and bifurcating monopolar neurones with varying lateral patterns of short fine fibres were common in the pond snail brain. Larger neurones have more complex and numerous branches originating from axons close to the perikarya than smaller ones. Stem processes originating on the cell body were observed on neurones above 30 in somal diameter. Possible sites for the location of chemical synapses were suggested. Functional types of neurones were difficult to separate on morphological grounds. Giant or very large neurones are small in number in pond snail ganglia, compared with medium or small neurones.The authors wish to thank Mr. Colin Atherton for photographic assistance and the U.K. Science Research Council for a grant to P. R. B.     

Possibilities of detecting the plasticity of monosynaptic excitable connections during the analysis of impulse activity of neurons using cross-correlation histograms and Cox's method

By the method of mathematical modelling, stationary irregular exogenous impulse activity of two monosynaptically connected neurones (N1 and N2; excitatory connection) was reproduced. The degree of impulse flows N1 and N2 dependence was estimated by the value P12, proportional to the height of crosscorrelation histogram peak, and by Cox's coefficient beta 12. The dynamics was studied of P12, beta 12 and of values P1* and P2*, proportional to mean interpulse intervals of N1 and N2, at changes of connection efficiency, neurones excitability and of influence of afferent synaptic bombardment on them. The following signs of modification of existing between N1 and N2 monosynaptic excitatory connection were established: 1) change of P12 in direction opposite to that of P1* and P2* change; 2) differently directed changes of beta 12 and P2* of postsynaptic N2.     

The morphology and electrophysiology of the neurones of the paired pedal ganglia of Lymnaea stagnalis (L.)

1. A morphological and electrophysiological map of the identifiable neurones and neuronal clusters of the paired pedal ganglia has been prepared. 2. Neuronal morphology was investigated using the fluorescent dye, Lucifer Yellow CH, whilst electrophysiological properties were studied using conventional intracellular recording techniques and the phase plane technique. 3. The paired pedal ganglia are largely symmetrical and giant neurones usually have contralateral homologues. 4. Neuronal clusters are also paired, but minor asymmetries, both of identifiable neurones and neuronal clusters have been found to exist. 5. These asymmetries are thought to be related to asymmetries of body form. 6. Most of the individually identifiable neurones possess obligatory axon branches which are invariant from one preparation to the next, but variant branches also occur. 7. Within the neuronal clusters, morphology appears to be more variable. 8. Individually identifiable neurones and neuronal clusters were characterized electrophysiologically according to the criteria of action potential shape, spontaneous activity pattern, electrical coupling and common synaptic inputs. 9. Homologous pairs of neurones usually have similar electrophysiological properties, as do those within clusters. 10. A number of wide-acting synaptic inputs have been identified on neurones of the pedal, buccal, visceral and parietal ganglia.     

Relation between fast and slow elemental synaptic potentials in command neurons in Helix lucorum taurica L

In experiments on a semi-intact snail preparation and a preparation of the snail isolated CNS, after spikes (Sp) evoked in presynaptic neurones by depolarizing current, not only rapid (R) EPSPs emerged in the command neurones of the defensive reaction of closing the pneumostome, but they were also followed by slow (S) EPSPs lasting over 2 min. For each single synaptic contact, the R and S EPSP amplitudes were in a good linear correspondence. In different synapses no direct connection was observed between R EPSP and S EPSP. It is suggested that R and S EPSPs may set in as a result of the action of different substances on the command neurones. Functional significance of S EPSPs with different amplitudes in different command neurones may consist in a prolonged specific preparation of the neurones for the action of stimuli.     

Statistical analysis and parsimonious modelling of dendrograms of in vitro neurones

The processes whereby developing neurones acquire morphological features that are common to entire populations (thereby allowing the definition of neuronal types) are still poorly understood. A mathematical model of neuronal arborizations may be useful to extract basic parameters or organization rules, hence helping to achieve a better understanding of the underlying growth processes. We present a parsimonious statistical model, intended to describe the topological organization of neuritic arborizations with a minimal number of parameters. It is based on a probability of splitting which depends only on the centrifugal order of segments. We compare the predictions made by the model of several topological properties of neurones with the corresponding actual values measured on a sample of honeybee (olfactory) antennal lobe neurones grown in primary culture, described in a previous study. The comparison is performed for three populations of segments corresponding to three neuronal morphological types previously identified and described in this sample. We show that simple assumptions together with the knowledge of a very small number of parameters allow the topological reconstruction of representative (bi-dimensional) biological neurones. We discuss the biological significance (in terms of possible factors involved in the determinism of neuronal types) of both common properties and cell-type specific features, observed on the neurones and predicted by the model. These authors contributed equally to this work.     

Ultrastructural studies on calcitonin gene-related peptide-, tachykinins- and somatostatin-immunoreactive neurones in rat dorsal root ganglia: Evidence for the colocalization of different peptides in single secretory granules

Summary Calcitonin gene-related peptide (CGRP)-, tachykinins- and somatostatin-immunoreactive neurones in rat dorsal root ganglia have been studied by means of single and double immunogold labelling techniques. Peptide-immunoreactive neurones are generally B- or C-type cells of small size, with well developed rough endoplasmic reticulum and scanty neurofilaments. In neurones classifiable as A₂-type cells, i.e. larger neurones with a lighter cytoplasm due to the presence of poorly developed Nissl bodies and numerous neurofilaments, only CGRP immunoreactivity was detected. Immunolabelled structures were identified as large (60–100 nm diameter), electron-dense, membranebounded p-type granules. They were observed only in neuronal cell bodies or in the intraganglionic portions of the axons. No granules immunoreactive to the antisera applied in this study were observed in non-neuronal cells. Immunostaining experiments with different combinations of the antisera revealed, in some cells, the presence of double immunolabelled granules; in particular localization of CGRP and tachykinins, CGRP and somatostatin, and tachykinins and somatostatin to single secretory granules was demonstrated. The finding that more than one peptide is localized to the same secretory granule supports the postulate that peptides are co-released upon nerve stimulation providing morphological support for physiological and pharmacological data demonstrating an interaction between different peptides in the modulation of synaptic activity.     

Photoaffinity labeling of ANF receptor in cultured brain neurones

A monoiodo derivative of rat atrial natriuretic factor (rANF) was shown to specifically bind to rat brain neurones in culture with low binding site capacity (10-20 fmoles per mg of protein) and high affinity (Kd = 50-100 pM). Several analogs of both rat and human ANF competed with 125I-rANF. No change in the number of binding sites was detected upon morphological differentiation of neurones in vitro. Finally a photoreactive derivative of 125I-rANF was prepared and photoaffinity labeling experiments carried out on cultured neurones. After reduction of disulfide bridges, a single band of Mr 60,000 was specifically labeled whereas without reduction, two labeled components of Mr 60,000 and 117,000 were detected.     

Neurotransmitters of giant neurons in the African giant snail, Achatina fulica Férussac. IV. Supplemental results

Following the previous works, we identified recently the twelve giant neurones in the ganglia of an African giant snail (Achatina fulica Férussac), by the pharmacological study of their sensitivities to putative neurotransmitters and derivatives, and by the morphological investigation of their axonal pathways due to the intracellular injection of Lucifer Yellow. The neurones studied were: TAN-2, TAN-3, BAPN, LPPN, LBPN and LAPN in the right parietal ganglion; RPeNLN and LPeNLN in the pedal ganglia; and d-LBAN, d-LBMN, d-LBCN and d-LBPN in the left buccal ganglion.     

Neuronal specialization of the motor cortex in normal rabbits and following destruction of the visual cortex

The activity of neurones of the anterolateral part of the motor cortex in food-acquisition behaviour was compared in two control rabbits and in three rabbits after the operation of bilateral ablation of the striatal cortex. In two of three operated rabbits the pattern of behavioural specialization lost considerably the specificity peculiar to the motor cortex (predominance of G-neurones activated in grasping of food), approaching (but not becoming identical) the pattern of specialization of the visual cortex neurones: the number of G-neurones decreased in a half, and the number of L-neurones (activated in connection with the acts of instrumental food-acquisition behaviour which animals were trained to in the experimental cage) was doubled. Changes of the activity were significantly less expressed in the third operated rabbit. The number of the neurones activated in food-acquisition behaviour in operated rabbits in comparison with the control ones was reduced in the upper layers of the cortex and increased in the lower layers. The resemblance is discussed of the basic processes of animals learning and behaviour recovery.