The serotonergic system in vertebrates and invertebrates has been a focus for over 50 years and will likely continue in the future. Recently, genomic analysis and discovery of alternative splicing and differential expression in tissues have increased the knowledge of serotonin (5-HT) receptor types. Comparative studies can provide useful insights to the wide variety of mechanistic actions of 5-HT responsible for behaviors regulated or modified by 5-HT. To determine cellular responses and influences on neural systems as well as the efferent control of behaviors by the motor units, preparations amenable to detailed studies of synapses are beneficial as working models. The invertebrate neuromuscular junctions (NMJs) offer some unique advantages for such investigations; action of 5-HT at crustacean NMJs has been widely studied, and leech and Aplysia continue to be key organisms. However, there are few studies in insects likely due to the focus in modulation within the CNS and lack of evidence of substantial action of 5-HT at the Drosophila NMJs. There are only a few reports in gastropods and annelids as well as other invertebrates. In this review we highlight some of the key findings of 5-HT actions and receptor types associated at NMJs in a variety of invertebrate preparations in hopes that future studies will build on this knowledge base.
We have established a model system of hormone action, in an Sf9 cell transfection system, using defined enhancer motifs and natural core promoters of metamorphosis-associated genes. The DR1 enhancer, that is an established DNA binding site for the ecdysone receptor/ultraspiracle heterodimer, was necessary for transcriptional activation by 20-OH ecdysone. For this activated transcription, a natural sequence closely 5′ to the TATA box is necessary. Cotreatment with juvenile hormone III strongly suppressed the steroid activation of transcription. However, in the absence of the sequence located closely 5′ to the TATA box, cotreatment with juvenile hormone instead increased transcription over that occurring due to 20-hydroxy-ecdysone alone. This sensitivity to activation by cotreatment with juvenile hormone could be transferred to a related, but otherwise unresponsive, hexamerin core promoter simply by transferring to the unresponsive promoter the five base transcription start site (ACAGT) from the responsive hexamerin gene. These are the first reports that the direction of JH action on 20-OH ecdysone-activated transcription can be reversed by removal of a sequence at the core promoter, and that modulatory action of juvenile hormone can be transferred to a different gene by transferring the transcription start site motif.
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