Extraglandular Innervation of the Salivary Cells of the Leech Haementeria Ghilianii: Neuronal Stimulation Elicits Gland-Cell Action Potentials and Secretion

Abstract

ABSTRACT Each salivary gland cell of Haementeria extends a single process, or ductule, anteriorly into the proboscis; secretory products are released at the ductule ending. Some ductules secrete into the lumen of the proboscis and others at the outer surface of its tip, more than 5 cm from the gland in large leeches. Depolarization of a gland-cell body elicits action potentials which appear to be conducted along the ductule to its ending. Electrical stimulation of the proboscis tip elicits action potentials in those ductules which end there, and the impulses are propagated to the cell body (approx. 5cms−1). Bathing the salivary glands in calcium-free saline causes spontaneous repetitive firing in the cell bodies and also elicits secretion at the proboscis tip (bathed in normal saline); the action potential thus appears to be a stimulus for secretion. A paired stomatogastric nerve, from the brain, enters the proboscis near its base. Cobalt-filling of the nerve shows numerous cell bodies in the brain and first body ganglion, and an intricate network of fibres and a cluster of stained cell bodies near its entry point in the proboscis. Repetitive stimulation of the stomatogastric nerve produces action potentials in certain gland cells, after a delay of at least 15 s, and also elicits secretion. The action potentials are initiated near the ductule tip, and are conducted to the cell body. The salivary glands themselves do not appear to be innervated. Application of acetylcholine (ACh), dopamine or octopamine (10−4mol 1−1) does not initiate secretion. Neither dopamine nor octopamine excites the gland cells but ACh produces a transient suprathreshold depolarization of the cell body and occasionally elicits 1–3 ductule spikes when applied to the proboscis tip. 5-Hydroxytryptamine (5-HT) produces secretion when applied to the proboscis but not when applied to the glands alone; it does not excite the cells, indicating that the action potential is not the only stimulus for secretion. 5-HT produces a depolarization, and increase in membrane resistance, in the cell body, and prevents the rapid adaptation of action potentials which occurs during maintained depolarization. 7.Electrophoretic analysis shows that the protein compositions of secretions at the proboscis tip and in the lumen are completely different, with the tip apparently secreting only two major proteins. These same two protein bands occur in the cytoplasm of certain gland-cell bodies which can be distinguished in living glands on the basis of size and degree of staining with Methylene Blue. 8.Following stimulation of the stomatogastric nerve, secretory products at the proboscis tip can be seen to emerge from discrete points which appear to be single ductule endings. This presents the possibility of studying excitation-secretion coupling in single cells.