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Extrahypothalamic peptidergic neurosecretion. II. Neurosecretion in the subfornical organ of Rana esculenta L.

In the subfornical organ of Rana esculenta, three basic structural elements can be demonstrated by light microscopic and immunohistological techniques used for the demonstration of products of the neurosecretory system. These elements are: (i) neurones and their processes, which the constituents of the subfornical organ proper, (ii) afferent axons of the preoptic nucleus, and (iii) subependymal cells with coarse processes. The vesicular inclusions of the two former structures correspond to the neurophysin vesicles with respect to their size, structure and reactivity. The vesicles of the subependymal cells belong to the same size class, possess a somewhat granular internal structure and react atypically after the application of the ultrahistochemical technique for the identification of neurophysin vesicles. Presumably, their content is a glycoprotein with a high proportion of cystine. The peptidergic axons of the preoptic nucleus projecting to the subfornical organ form neuroneuronal synapses.

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Monoclonal antibodies against Nereis virens brain homogenates as probes for isolation and in situ detection of new neurosecretions.

The brain of Nereis contains 26 ganglionic nuclei which produce numerous neurosecretions. Only a few of them have been characterized. The production of monoclonal antibodies was adopted as an approach to discover unknown neurosecretions. Monoclonal antibodies produced against Nereis virens brain homogenates were selected using stepwise ELISA tests first with brain homogenates, then with brain neurosecretions. Eight antibodies specific for Nereis neurosecretions were selected. The results are illustrated with one of these monoclonal antibodies which was directed against a major peak after HPLC purification of brain neurosecretions. This antibody was subsequently used for the in situ detection of recognized epitope(s) in the brain and ventral nerve cord cells.

Animals↗

Is hyperosmotic neurosecretion from motor nerve endings a calcium-dependent process?

Spontaneous liberation of neurotransmitter quanta is strongly affected by the osmotic pressure of the extracellular fluid. Elevation of the osmolarity by 20-30% increases the rate of release from motor nerve endings by more than one order of magnitude. In this respect the neuromuscular junction resembles some other secretory systems. The mechanism of this hyperosmotic neurosecretion is not yet understood; extracellular calcium ions are not directly responsible, since this effect can be produced in their absence. Recently, it has been suggested that the liberation of neurotransmitter is regulated by the intracellular concentration of free calcium ions. We have therefore examined the hypothesis that hyperosmotic neurosecretion originates from an increase in internal calcium concentration ([Ca]in). At the frog neuromuscular synapse however, it is impossible at present to estimate directly free [Ca]in; hence we used an indirect technique, which is based on two assumptions; first, the frequency of the miniature endplate potentials (m.e.p.p.s.) reflects free [Ca]in. Second, the movement of calcium ions across the presynaptic membrane is governed by the electrochemical gradient, and by the calcium conductance (g(Ca)). If hyperosmotic neurosecretion is caused by an increase in [Ca]in, then increasing g(Ca), under reversed electrochemical gradient for the calcium should cause a reduction in the effect of hyperosmotic stress on transmitter release. We report that hyperosmotic neurosecretion is dependent on [Ca]in.

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