E. W. Roubos

Synaptology of the central nervous system of the freshwater snail Lymnaea stagnalis (L.), with particular reference to neurosecretion.

SummaryIn the central nervous system (CNS) of the freshwater snail Lymnaea stagnalis 3 types of interneuronal contacts can be distinguished electron-microscopically, viz. true synapses, “synapse-like structures” (SLS), and “spinules”. Use of the electron microscope specimen tilting stage reveals numerous true synapses. Both “terminal” and “en passant” contacts occur on neurones and on glial cells. Furthermore “bigeminal” synapses are present. Complex (combined) convergent and divergent synaptic arrangements are found. On the basis of the morphology of presynaptic vesicles 7 types of true synapses can be discerned. Histochemical data on the contents of the vesicles are lacking. However, vesicle morphology suggests that type IV is aminergic and type VII cholinergic. Terminal and en passant SLS may penetrate deeply into neuronal somata and large axons, and into glial cells. A cluster of synaptic vesicles is present in the presynapse-like element. Spinules (spine-coated “evagination-invagination” specializations of the plasma membranes of 2 adjacent neuronal elements) are observed between somata, between axons, and between soma and axon.The neurosecretory Light Green Cells (LGC) and Caudo-Dorsal Cells (CDC) receive complex synaptic input. Type V true synapses, 2 types of SLS, and spinules contact the LGC. The complex morphology of the relationship between type A SLS and LGC, studied in serial sections, reveals that adjacent glial cells are also contacted by type A SLS. Type II true synapses, 3 types of SLS, and spinules are identified on the CDC.The validity of the methods of identification and classification of interneuronal contacts in the CNS of L. stagnalis, as well as the role of these contacts in the regulation of the activity of “ordinary” neurones, neurosecretory cells, and glial cells is discussed.

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

SummaryThe 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.The physiological significance of the neuronal and non-neuronal control mechanisms which regulate CDC activity is discussed.

Neurosecretion in the basommatophoran snail Bulinus truncatus (Gastropoda, Pulmonata).

SummaryThe neurosecretory system of the freshwater snail Bulinus truncatus was investigated. With the Alcian blue-Alcian yellow (AB/AY) staining method at least 10 different types of neurosecretory cells (NSC) were distinguished in the ganglia of the central nervous system. The differences in staining properties of the NSC — with AB/AY the cells take on different shades of green and yellow — are borne out at the ultrastructural level: the NSC types contain different types of neurosecretory elementary granules.The neurosecretory system of B. truncatus is compared to that of Lymnaea stagnalis, the species which has received the most attention among the pulmonates. It appears from the comparison that the systems of both species show many similarities, although some differences are also apparent.

Ultrastructural demonstration of secretion by exocytosis in rat pinealocytes with the use of the tannic acid method

SummaryIn the rat pineal gland the mechanism of release of secretory material was studied ultrastructurally after incubating tissues in Ringer solution containing tannic acid. The results indicate that pinealocytes release the contents of secretory vesicles into the extracellular space via exocytosis, a phenomenon that has not been visualized previously in this cell type. This finding may reflect release of polypeptides by the pineal gland.

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

SummaryThe neurosecretory Dark Green Cells (DGC) in the pleural and parietal ganglia of the freshwater snail Lymnaea stagnalis seem to be involved in osmoregulation. Previous experiments have indicated that changes of the osmolality of the environment induce activity changes of the DGC. Furthermore, it was shown that information on environmental osmolality reaches the DGC via the blood.In the present study right pleural and parietal ganglion complexes were cultured for 3 days in vitro under different osmotic conditions. Quantitative electron microscopy revealed that, compared with the control osmolality (130 mOsm/kg H2O), osmolalities of 160 and 190 mOsm/kg H2O caused a reduced synthesis and an increased storage of neurohormone in the DGC. Apparently, the activity of the DGC depended on the osmotic pressure of the medium. It is proposed that in vivo the osmotic pressure of the blood (which is related to the osmolality of the environment) regulates DGC activity.

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

SummaryThe 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.

Roles of Calcium and cAMP in Biosynthesis and Release of the Ovulation Hormone of the Freshwater Snail Lymnaea stagnalis

Because of their large size and easy accessibility for various morphological, physiological, and pharmacological experimental approaches, neurons in the central nervous system of molluscs are very suitable objects for studies of fundamental problems in neurobiology. The bag cells of the sea hare Aplysia californica [1,2] and the caudo-dorsal cells (CDC) of the freshwater snail Lymnaea stagnalis [3,4] belong to the most extensively studied neuronal systems of molluscs. Investigations of these cell types have provided considerable insight into the significance of peptides in neural and neuroendocrine communication, not only as to their diverse functions, but also with respect to the mechanisms that control peptide biosynthesis and release. In this paper we will consider recent neurobiological studies of the CDC with particular reference to functional-morphological aspects.