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Modulatory information plays a key role in the expression and the ontogeny of motor networks. Many developmental studies suggest that the acquisition of adult properties by immature networks involves their progressive innervation by modulatory input neurons. Using the stomatogastric nervous system of the European lobsterHomarus gammarus, we show that contrary to this assumption, the known population of projection neurons to motor networks, as revealed by retrograde dye migration, is established early in embryonic development. Moreover, these neurons display a large heterogeneity in the chronology of acquisition of their full adult neurotransmitter phenotype. We performed retrograde dye migration to compare the neuronal population projecting to motor networks located in the stomatogastric ganglion in the embryo and adult. We show that this neuronal population is quantitatively established at developmental stage 65%, and each identified projection neuron displays the same axon projection pattern in the adult and the embryo. We then combined retrograde dye migration with FLRFamide-like, histamine, and GABA immunocytochemistry to characterize the chronology of neurotransmitter expression in individual identified projection neurons. We show that this early established population of projection neurons gradually acquires its neurotransmitter phenotype complement. This study indicates that (1) the basic architecture of the known population of projection inputs to a target network is established early in development and (2) ontogenetic plasticity may depend on changes in neurotransmitter phenotype expression within preexisting neurons rather than in the addition of new projection neurons or fibers.

Ontogeny of Modulatory Inputs to Motor Networks: Early Established Projection and Progressive Neurotransmitter Acquisition