Environmental osmolarity and neurosecretory neurones in Lymnaea stagnalis (L.).

Cell body volume and Alcian blue-Alcian Yellow staining properties of neurosecretory neurones in the brain of Lymnaea stagnalis were compared for snails kept in de-ionised water and standard tapwater. In the same experiment, the ionic content of the blood, blood volume and body weight and environmental ionic composition were measured. Five days of immersion in de-ionised water resulted in significant decreases in body weight, blood volume and blood, Na+ and Cl- concentrations but no change in blood Ca2+, K+,and HCO3- concentrations, compared with controls. No consistently significant differences across the 5 day period were found in cell body volumes for Dark Green Cells, Yellow Cells or Light Green Cells (used as a control) when these volumes were compared for large numbers of cells from snails kept in de-ionised water and standard tapwater. However, the number of Yellow Cells which could be counted in snails kept in de-ionised water was lower than the number from standard tapwater by day 2 of the experiment and lower for Yellow-green Cells by day 5. We interpret this lower number to be the result of depletion of Alcian blue-Alcian Yellow stained neurosecretory material in these cells which made them impossible to distinguish. This was confirmed by examination of visceral Yellow Cells which could be identified on the basis of known location close to the visceral-right parietal connective.     

Regulation of neurosecretory activity in the freshwater pulmonate Lymnaea stagnalis (L.) with particular reference to the role of the eyes

The process of neurosecretion in the Caudo-Dorsal Cells (CDC) of the freshwater snail Lymnaea stagnalis, which produce an ovulation hormone, shows a diurnal rhythmicity. Synthesis, transport and release of the neurosecretory material (NSM) is high during the evening and the early night and low during the rest of the day, while storage of NSM mainly occurs during the daytime. In the present study the role of the eyes in the regulation of the CDC-rhythm was investigated. During a 24-hr period, at time intervals of 6 hrs, cerebral ganglia, which contain CDC, of blinded and control snails (5 per group) were fixed and the CDC were studied with quantitative electron microscopical methods. The CDC of the controls showed a distinct diurnal thythmicity. Blinding, on the other hand, clearly affected this rhythmicity. The results indicate that after blinding the circadian CDC-rhythms of individual snails are no longer synchronous with each other ("interanimal desynchronization"). It is suggested that the rhythm of CDC neurosecretory activity is synchronized by the natural light/dark cycle via the eyes. The information from the eyes probably reaches the CDC via a nervous pathway. True snyapses and three types of synapse-like structures were found on the CDC. Their role in the regulation of CDC-activity is discussed. The effect of blinding is specific for the CDC; blinding does not influence the diurnal rhythmicity of another type of cerebral neurosecretory cells, the Light Green Cells (LGC). The CDC within a cluster act synchronously. This synchrony does not depend upon the presence of the eyes. Some structures which may be involved in establishing this synchrony, such as subsurface cisterns, desmosome-like structures and "specific release sites", are described.     

Electron microscopic investigation of a giant neuron identified in the right parietal ganglion of Lymnaea stagnalis (L)

The ultrastructure of a giant neuron (GRP1) identified in the right parietal ganglion of Lymnaea stagnalis was examined. The results suggest that GRP1 is a neurosecretory cell. The perikaryon is characterized by many neurosecretory-like granules of variable electron density. The average diameter (longer axis) of the granules was 1850 A. Furthermore highly-developed rough endoplasmic reticulum and Golgi complex were observed in the cytoplasm. Synapse-like structures were often found on the somatic membrane, but typical, true synaptic endings were not seen.     

The anatomy of neurosecretory neurones in the pond snail Lymnaea stagnalis (L.).

The anatomy of three neurosecretory cell types in the central nervous system (c.n.s.) of the gastropod mollusc Lymnaea stagnalis (L.)- the Dark Green Cells, Yellow Cells and Yellow-green Cells-has been studied by using bright and dark field illumination of material stained for neurosecretion by the Alcian Blue-Alcian Yellow technique. The neuronal geometry of single and groups of neurosecretory cells of the various types has been reconstructed from serial sections, and the likely destination of most of their processes has been determined. Dark Green Cells are monopolar, occur exclusively within the central nervous system (c.n.s.), have few or no branches terminating in neuropile, and send axons to the surface of the pleuro-parietal and pleuro-cerebral connectives. The majority of Dark Green Cell axons however (80-85%), project down nerves which innervate ventral and anterior parts of the head-foot, the neck and the mantle. Dark Green Cell axons can be found in small nerves throughout these areas, and may terminate in a find plexus of axons on the surfaces of the nerves. Since previous experimental work has shown that the Dark Green Cells are involved in osmotic or ionic regulation, these results suggest that the target organ of the Dark Green Cells may be the skin. Yellow Cells occur both within and outside the c.n.s. They are usually monopolar, but can be bipolar. They have several axons which normally arise separately from a single pole of the cell body, or close to it. One or more processes leave the cell proximal to the point where separate axons arise, and may run unbranched for some distance through neuropile before terminating in fine brances and blobs of various sizes. These branches may release hormone inside the c.n.s. Yellow-green Cells are mono-, bi- or multi-polar, and like the Yellow Cells are found both within and outside the c.n.s. Some Yellow-green Cells, though not all, have projections which terminate in neuropile in fine branches and blobs. Yellow-green Cell bodies which occur in nerves can project back along the nerve into the c.n.s. The axons of Yellow Cells and Yellow-green Cells project to release sites in various ways. Some project into the connective tissue shealth of the c.n.s., which serves as a neurohaemal organ, either directly through the surface of a ganglion, or from the pleuro-cerebral or pleuro-parietal connectives. Other axons leave the c.n.s. via nerves leaving the left and right parietal and visceral ganglia; projections into the intestinal, anal, and internal right parietal nerves being most numerous. Axons which may be from either, or both Yellow Cells and Yellow-green Cells, can be found along the entire unbranched lengths of these nerves, and in subsequent branches which innervate organs lying in the anterior turn of the shell. All of these orgnas are closely associated with the lung cavity...     

Neuronal and non-neuronal control of the neurosecretory Caudo-Dorsal cells of the freshwater snail Lymnaea stagnalis (L.)

The cerebral ganglia of the freshwater snail Lymnaea stagnalis contain two clusters of neurosecretory Caudo-Dorsal Cells (CDC). These cells produce a neurohormone which stimulates ovulation. Ganglion transplantation and quantitative electron microscopy show that neuronal isolation of the cerebral ganglia complex (CCC) results in an activation of the CDC. It was, therefore, concluded that the CDC are controlled by an inhibitory neuronal input originating outside the cerebral ganglia. Ultrastructural studies on synaptic degeneration in the CCC suggest that this input reaches the CDC via a special type of synapse-like structure, the type C-SLS.Furthermore, transplantation of CCC into acceptor snails leads to a reduced release and an increased intracellular breakdown of neurohormone in the CDC of the nervous system of the acceptors. It is supposed that these phenomena are caused by the release of an (unknown) factor from the transplanted CCC. Special attention was given to the formation and degradation of a peculiar type of neurohormone granule, the large electron dense granule.     

Structure and electrophysiological properties of bursting neurosecretory cells in a peripheral sensory ganglion of the pond snail Lymnaea stagnalis

A group of peripheral neurosecretory oscillating neurons belonging to the type of parabolic bursters, were identified in the osphradium (peripheral putative chemosensory organ) of the pond snail Lymnaea stagnalis. The cells are unipolar, their process ramifies and terminates in the nerve. Applications of 5-HT caused long-lasting bursts with significantly increasing duration and frequency of spikes. GABA and FMRFamide inhibited the activity of these cells.     

Electrotonic coupling within a cluster of neurosecretory endogenous oscillators in Lymnaea stagnalis (L.)

• 1.1. Double intracellular and extracellular recordings from cell bodies and axons were made to study the interactions between the neurosecretory “Light Yellow” bursting pacemaker cells (LYC) in the right parietal ganglion of Lymnaea stagnalis.
• 2.2. The LYC are interconnected by low efficiency, non-rectifying electrotonic junctions, transmitting low frequencies only.
• 3.3. Often bursts in different cells coincide; apparently the junctions are responsible for this coherence.
• 4.4. It is inferred that the coupling serves to bring about a pulse-wise release of the cluster's secretory product.

     

An ultrastructural in vitro study on the regulation of neurosecretory activity in the freshwater snail Lymnaea stagnalis (L.) with particular reference to Caudo-dorsal cells

The neurosecretory Caudo-Dorsal Cells (CDC) in the cerebral ganglia of the freshwater pulmonate snail Lymnaea stagnalis produce an ovulation stimulating hormone. Previously it has been shown that neuronal and non-neuronal inputs are involved in the regulation of their activity. The degree of autonomy of these cells has been investigated by studying with morphometric methods the ultrastructure of CDC maintained in vitro. CDC of isolated cerebral ganglia which were cultured for 7 days show a considerable rate of synthesis, transport and release of neurohormone. Apparently these processes can proceed in the absence of neuronal and hormonal inputs from outside the cerebral ganglia. Completely isolated CDC, however, do not show neurosecretory activity in vitro; active Golgi zones, indicating the formation of neurosecretory elementary granules, are absent from such cells. Isolation does not seem to affect general cell functions such as protein synthesis and respiration. It is suggested that a neuronal input, originating within the cerebral ganglia, is necessary for the stimulation of CDC neurosecretory activity. Techniques are described for the isolation and culture of neurosecretory cells of L. stagnalis.     

Morphometric in vitro analysis of the control of the activity of the neurosecretory dark green cells in the freshwater snail Lymnaea stagnalis (L.)

Summary The intracellular localization of calcium by means of cytochemical techniques was studied in smooth muscle cells of mouse intestine. When the lead acetate method according to Carasso and Favard (1966) was used calcium was found in mitochondria and sarcoplasmic reticulum and occasionally between the myofilaments. The active ATP-dependent accumulation of calcium into cell structures was investigated by the oxalate method (Heumann and Zebe, 1967). After appropriate treatment the only structures of smooth muscle cells which contained calcium oxalate (identified by microprobe analysis) were elements of the sarcoplasmic reticulum.The results are discussed in relation to the role of calcium in the control of muscle activity during the contraction-relaxation cycle.The electron probe microanalysis was carried out at SIEMENS (Berlin) in collaboration with Dr. von Muschwitz. I thank Miss M. Schlatter for her skillful assistance. The investigation was supported by the Deutsche Forschungsgemeinschaft.     

A histoenzymatic investigation of galactogen synthesis in the albumen gland of Helix pomatia and Lymnaea stagnalis

Summary A histoenzymatic investigation of Galaotogen synthesis has been attempted in the albumen gland of Helix pomatia and Lymnaea stagnalis. Using histoenzymatic methods the authors succeeded in finding only two enzymes (E.C. 2.7.7.9.=uridylyltransferase and E. C. 5.1.3.2.=epimerase) demonstrated by biochemical assays and isolated. They therefore assume that there may be other metabolic pathways in addition to those demonstrated by biochemical methods.     

Ultrastructural study of the follicle cells in the freshwater gastropod Viviparus viviparus L.

Summary The morphology and the role of the follicle cells of Viviparus viviparus were examined by means of light and electron microscopy. The follicle cells appear to contain glycogen and fat, and often lysosomes or heterogeneous inclusions. Therefore, they seem to be active in phagocytosis and storage. They are probably involved in the nutrition of the oocyte. Their role in the formation of a selectively permeable barrier is discussed.The authors thank Drs. H.H. Boer, M. de Jong-Brink and J. Wijdenes of the Free University of Amsterdam for their assistance in the translation of this paper     

Periodic and oscillatory firing patterns in identified nerve cells of Lymnaea stagnalis L

Firing patterns in identified neurons of Lymnaea stagnalis L. were analyzed by various mathematical methods including spike density function (SDF), interspike-interval histograms (ISI), Fourier transform and correlation analysis. Input-3 (IP3) events observed in most of the neurons of the respiratory regulatory system caused prominent changes in the firing frequency of the cells. Similarly, quasiperiodic firing patterns were observed in the neurons of buccal ganglia controlling feeding behavior. Apart from the known periodic patterns a fine oscillation of firing rate was observed in a large number of neurons in the visceral and parietal ganglia. The frequency of this oscillation varied between 0.2 and 0.4 Hz. The most obvious oscillatory patterns were found in the A-cells presumably resulted by periodically appearing synaptic excitation. Moderate intracellular hyperpolarizing current injection, low-Ca/high-Mg saline and application of d-tubocurarine failed to abolish the slow oscillations. Application of Ca-channel blocker cadmium, however, completely eliminated the oscillation in a reversible manner.