This study demonstrates the presence of a relatively extensive but previously unrecognized nervous system in embryonic stages of the opisthobranch mollusc Aplysia californica. During the trochophore stage, two pairs of cells were observed to be reactive to antibodies raised against the neuropeptides FMRFamide and EFLRIamide. These cells were located in the posterior region of the embryo, and their anterior projections terminated under the apical tuft. As the embryos developed into veliger stages, serotonin-like immunoreactive (LIR) cells appeared in the apical organ and were later observed to innervate the velum. Also, aldehyde-induced fluorescence indicative of catecholamines was present in cells in the foot, oral, and possibly apical regions during late embryonic veliger stages. Just before the embryo hatches as a free-swimming veliger, additional FMRFamide-LIR and catecholamine-containing cells appeared in regions that correspond to the ganglia of what will become the adult central nervous system (CNS). Neurons and connectives that will contribute to the adult CNS appear to develop along the pathways that are pioneered by the earliest posterior FMRFamide-LIR cells. These observations are consistent with the hypothesis that, besides their presumed roles in the control of embryonic behaviors, some elements may also guide the development of the CNS. Embryonic nervous systems that develop prior to and outside of the adult CNS have also been reported in pulmonate and prosobranch species of molluscs. Therefore, the demonstration of early developing neurons and their transmitter phenotypes in A. californica presents new opportunities for a better understanding of the ontogeny and phylogeny of both behavioral and neuronal function in this important model species.
Larval molluscs commonly use ciliated vela to swim and feed. In this study we used immunohistochemistry to demonstrate innervation of velar cilia and muscles by monoaminergic and peptidergic fibres in the caenogastropod, Ilyanassa obsoleta. Photoelectric recordings from pre-oral cilia on isolated pieces of velum revealed that serotonin increased, whereas catecholamines (dopamine and norepinephrine) decreased beat frequency at concentrations of 10-6 to 10-9 mol l-1. Catecholamines also increased the frequency of momentary, isolated arrests of pre-oral cilia, but failed to suppress beating of the post-oral cilia at these concentrations. The neuropeptides, FMRFamide and Leu-enkephalin, did not affect the frequency of ciliary beating or of isolated ciliary arrests, but did induce numerous muscular contractions, which were accompanied by sustained ciliary arrests. In terms of whole animal behaviour, serotonin caused larvae to concentrate toward the top of a water column and to increase feeding, whereas catecholamines caused larvae to concentrate toward the bottom of a water column and decrease feeding. Monoamine analogues which facilitated or opposed the effects of synthetic transmitters on larval behaviour, further suggested that these transmitters are released endogenously to control velar function. Finally, applications of peptides to whole larvae caused increased frequency of locomotory arrests. Together these findings demonstrate several potential roles for the nervous system in controlling larval behaviour in gastropods.