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Organization of the serotonergic innervation of the feeding (buccal) musculature during the maturation of the pond snail Lymnaea stagnalis: A morphological and biochemical study

The serotonergic innervation of the buccal musculature responsible for feeding (radula protraction) was investigated during the maturation of the pond snail, Lymnaea stagnalis L., applying light and electron microscopic immunohistochemistry and biochemical approaches. According to epifluorescence and laser confocal microscopy, the first 5‐HT‐like‐immunoreactive (5‐HTLIR) processes appeared on the surface of the musculature at the postmetamorphic E80% embryonic stage. Until hatching, the innervation continued to increase in density, showing axon arborizations with projections into the deeper muscle levels. An adult‐like pattern of 5‐HTLIR innervation appeared at P2–P3 juvenile stages. At the ultrastructural level, close (16–20 nm) but mostly unspecialized neuromuscular contacts were formed by both unlabeled and 5‐HTLIR axon profiles from the E80% embryonic stage. Labeled processes were also found located relatively far from the muscle cells. An HPLC assay showed a gradual increase of the 5‐HT level in the buccal mass during development. The buccal mass was characterized by a single‐component high‐affinity 5‐HT uptake system, and 5‐HT release could be evoked by 100 mM K+ and blocked in Ca2+‐free medium. It is suggested that 5‐HT plays a wide modulatory role in the peripheral feeding system and is also involved in the functional maturation of the muscle system. J. Comp. Neurol. 520:315–329, 2012. © 2011 Wiley Periodicals, Inc.