Dopamine-synthesizing neurons include the putative H-cell homologue in the moth Manduca sexta

The catecholamine dopamine (DA) plays a fundamental role in the regulation of behavior and neurodevelopment across animal species. Uncovering the embryonic origins of neurons that express DA opens a path for a deeper understanding of how DA expression is regulated and, in turn, how DA regulates the activities of the nervous system. In a well-established insect model, Manduca sexta, we identified the putative homologue of the embryonic grasshopper "H-cell" using intracellular techniques, laser scanning confocal microscopy, and immunohistochemistry. In both species, this neuron possesses four axons and has central projections resembling the letter H. The H-cell in grasshoppers is known to be derived from the midline precursor 3 cell (MP3) and to pioneer the pathways of the longitudinal connectives; in Drosophila, the H-cell is also known to be derived from MP3. In the current study, we demonstrate that the Manduca H-cell is immunoreactive to antibodies raised against DA and its rate-limiting synthetic enzyme, tyrosine hydroxylase (TH). In larvae and adults, one DA/TH-immunoreactive (-ir) H-cell per ganglion is present. In embryos, individual ganglia contain a single midline TH-ir cell body positioned along side its putative sibling. Such observations are consistent with the known secondary transformation (in grasshoppers) of only one of the two MP3 progeny during early development. Although a hallmark feature of invertebrate neurons is the fairly stereotypical position of neuronal somata, we found that the H-cell somata can "flip-flop" by 180 degrees between an anterior and posterior position. This variability appears to be random and is not restricted to any particular ganglion. Curiously, what is segment-specific is the absence of the DA/TH-ir H-cell in the metathoracic (T3) ganglion as well as the unique structure of the H-cell in the subesophageal ganglion. Because this is the first immunohistochemical study of DA neurons in Manduca, we have provided the distribution pattern and morphologies of dopaminergic neurons, in addition to the H-cells, within the ventral nerve cord during development.