Co-existence of immunoreactivity to anti-dopamine,anti-serotonin and anti-vasotocin in the cerebral giant neuron of the pond snail Lymnaea stagnalis

Summary Consecutive sections of certain neurons in the central ganglia of the pond snail Lymnaea stagnalis appear to be immunoreactive to anti-dopamine and anti-serotonin. The Cerebral Giant Neurons stain in addition with antivasotocin. The observations indicate the presence of two biogenic amines within the same neuron and in addition their co-existence with a biologically active peptide.     

Studies on the oviposition and longevity ofIlione albiseta [Dipt.: Sciomyzidae] — Potential biological control agent of liver fluke

The effect of 5 constant temperatures (10, 14, 17, 20 and 23°C) on the oviposition and longevity ofIlione albiseta was investigated. Most eggs were laid at 14–17°C and mean oviposition period declined progressively above and below 14°C. There was no significant difference between oviposition rates or preoviposition periods at each constant temperature, but the mean number of days between egg laying for each female was significantly greater at 10 °C than at 17, 20 and 23 °C than at 20 °C. The percentage of infertile eggs laid ranged from 9.2% at 23 °C to 17.9 % at 20 °C and these eggs tended to be laid at the beginning and end of each oviposition period. A possible association between sex ratio of the emergent adult and temperature is also discussed.      

Functional morphology of the neuroendocrine sodium influx-stimulating peptide system of the pond snail,Lymnaea stagnalis,studied by in situ hybridization and immunocytochemistry

Summary The functional morphology of the neuroendocrine system producing sodium influx-stimulating (SIS) peptide in the pond snail, Lymnaea stagnalis, was studied by in situ hybridization and immunocytochemistry. The SIS-peptide, which is 76 amino acids long, stimulates sodium uptake from the ambient medium. Two synthetic DNA probes were used for in situ hybridization. The nucleotide sequences were chosen from the cDNA structure; they encode amino acids 8–17 and 64–73, respectively. SIS-peptide sequences 10–20 and 67–76 were synthesized and antibodies were raised to them and affinity-purified. In addition to these antibodies, a monoclonal antibody raised to a bioactive, high-pressure liquid chromatography (HPLC)-purified brain extract was used for immunocytochemistry. Paraffin sections of central nervous systems and of whole snails were studied. The SIS-peptide system could be identified as the previously described yellow cell (YC) system by comparing alternate sections treated with the DNA probes, stained with the antibodies, or stained with alcian blue-alcian yellow. SIS-peptide neurons (45) occur in the ganglia of the visceral ring and in the proximal parts of visceral nerves. Axons run in the nerves of these and in several nerves of other ganglia. Numerous axon branches penetrate the perineurium forming a vast central neurohemal area. The SIS-peptide system innervates the pericardium, the nephridial gland, the reno-pericardial canal, the ureter, the spermoviduct and gonadal acini, the anterior aorta, the ventral buccal artery, and the penis protractor muscle. The morphology of the system is discussed in relation to the process of sodium ion uptake from the ambient medium and from pro-urine, and to that of regulating blood pressure. In the central nervous system and other organs, neurons and axons not labeled with the DNA probes, but immunoreactive to one or two of the antibodies, were observed. It seems unlikely that these elements are functionally related to the SIS-peptide system.     

Chronic toxicity of three insecticides (carbaryl,fenthion and lindane) in the freshwater snail Lymnaea stagnalis

A chronic intoxication with carbaryl, fenthion and lindane was induced in young snails. The parameter k of the von Bertalanffy's equation showed clearly the growth changes induced by these insecticides. In all cases the fecundity of intoxicated snails was reduced. Among these three insecticides, lindane was the most toxic, carbaryl the least.     

Unusual distribution of tubulin isoforms in the snail Lymnaea stagnalis

Immunocytochemistry and Western blotting techniques demonstrated that the nervous system and foot of the pond snail Lymnaea stagnalis are rich sources of tubulin, which can be extracted and assembled in vitro in the presence of taxol. Various broad-spectrum antibodies raised against -tubulin and -tubulin yielded qualitatively similar results. One monoclonal antibody to trypanosome -tubulin, however, labelled -tubulin more strongly on both probed sections and Western blots. Cytochemistry and immunoblotting revealed that tyrosinated tubulin constitutes a large proportion of total -tubulin in locomotor cilia of the foot and in axons of the nervous system. Detyrosinated tubulin also appeared to be abundant in the foot cilia but only a very faint band of detyrosinated tubulin was found on protein blots extracted from the central ganglia, and staining was barely detectable in central ganglia or peripheral nerves. Similarly, acetylated tubulin appeared to be abundant in foot cilia, but Western blotting indicated only low levels of acetylated tubulin in the nervous system. Immunocytochemistry indicated that, while most neurons possessed little or no acetylated tubulin, a small number of axons contained significant amounts of this isoform. Thus, while a large amount of tubulin was expected in the nervous system and locomotor cilia of L. stagnalis, the observed distribution of isoforms was unanticipated. Specifically, neurons of other organisms have generally been reported to contain substantial amounts of both detyrosinated -tubulin and acetylated -tubulin. Our results indicate that such findings cannot be generalized across all species. L. stagnalis, with its well studied nervous system and unusual distribution of tubulin isoforms, may prove to be particularly useful for studying the roles of tubulin isoforms in microtubule function and cell activity.     

Ultrastructural immunocytochemical evidence for peptidergic neurotransmission in the pond snail Lymnaea stagnalis

Summary Three neuronal systems of the pond snail Lymnaea stagnalis were immunocytochemically investigated at the ultrastructural level with the unlabeled peroxidase-antiperoxidase technique. Preliminary electrophysiological and cell-filling investigations have shown that a cluster of neurons which reacts positively with an antiserum against the molluscan cardio-active peptide FMRFamide, sends axons to the penis retractor muscle. In this muscle anti-FMRF-amide (aFM) positive axons form neuro-muscular synapses with (smooth) muscle fibers. The morphological observations suggest the aFM immunoreactive system to be involved in peptidergic neurotransmission. In the right parietal ganglion a large neuron (LYAC) is penetrated by aFM positive axons which form synapse-like structures (SLS) with the LYAC. The assumption that the SLS represent the morphological basis for peptidergic transmission is sustained by the observation that iontophoretical application of synthetic FMRFamide depolarizes the LYAC. The axons of a group of pedal anti-vasopressin (aVP) positive cells run in close vicinity to the cerebral ovulation (neuro-)-hormone producing cell system (CDC system) Synapses or SLS between the two systems were not observed. The fact that (bath) application of arg-vasopressin induces bursting in the CDC, may indicate that the vasopressin-like substance of the aVP cells is released non-synaptically.     

Sensory cells in the head skin of pond snails

Summary Several types of receptor endings were identified with scanning electron microscopy and silver-impregnation techniques in the skin of the tentacles, lips, dorsal surface of the head and mouth region of the pond snails Lymnaea stagnalis and Vivipara viviparus. Sensory endings at the tips of dendrites of primary receptor neurones, scattered below the epithelium, differ in structure, i.e., the endings exposed to the surface of the skin possess different proportions of cilia and microvilli, which vary in number, length, and packing. Type-I endings have microvilli and a few (1–5) cilia, 5–12 m in length. Type-2 endings have abundant (20–40), interwoven long (9–12 m) cilia and random microvilli. Type-3 endings show typical packing of 10–25 cilia in the form of bundles or brushes. They may be composed either of long (9–18 m) or short (2–7 m) cilia, or of both long and short ones. Microvilli here are absent. Type-4 endings have only microvilli. Two other types of skin receptors do not extend their sensory endings to the surface and can be indentified only in silver-stained preparations. Type-5 endings are branching dendrites of skin receptors cells that terminate among epithelial cells. In type-6, the sensory endings also terminate among epithelial cells but their cell bodies are located outside of the skin. In both species all skin regions examined possess the receptors of all six types differing only in their relative proportion. Possible functional roles of different receptors are discussed.     

Interaction of photoperiod and nutritive state in female reproduction of Lymnaea stagnalis

Summary

A study was made of the interaction of the photoperiod and the availability of food in influencing egg laying of the freshwater snail Lymnaea stagnalis. At a long day photoperiod (LD = 16 h light/8 h dark) egg laying of fed snails had increased compared with that of fed animals kept at a medium day photoperiod (MD = 12 h light/12 h dark). In MD snails oviposition ceased within a week of the beginning of a starvation period. This is most probably due to a reduction in the activities of the neuroendocrine caudodorsal cells which secrete an ovulation hormone. In contrast, in starved LD snails a low rate of ovipository activity continued, indicating that a lowered frequency of caudodorsal cell release cycles occurred under these conditions. The decreased mean size (number of eggs) of the egg masses in starved LD snails indicates that the activities of the endocrine dorsal bodies, which control vitellogenesis and synthetic activities in the female accessory sex organs, had decreased.

All MD snails survived, but nearly all LD snails died during the course of the experiment. Determinations of the mantle glycogen stores of LD snails suggest that the high mortality of LD snails is due to exhaustion of the animal's energy reserves.     

Regulation of secretory activity in the albumen gland of the pulmonate snail Lymnaea stagnalis (L.)

The secretory activity of the albumen gland of the freshwater snail Lymnaea stagnalis was studied morphometrically (both at the light- and at the electron-microscope level) and biochemically under the following experimental conditions: (1) glandular tissue was implanted into acceptor snails and the glandular activity of the implants was compared to that of the glands of the donors and acceptors; (2) glandular activity was measured at various periods after oviposition; and (3) the activity was measured during a 24 h cycle (diurnal activity). The results indicate that cellular release of secretion material is regulated by a nervous mechanism, whereas synthetic activity is under hormonal control.