Putative circadian pacemaker cells in the antenna of the hawkmoth Manduca sexta

Antennal sensory neurons in the fruit fly Drosophila melanogaster express circadian rhythms in the clock gene PERIOD (PER) and appear to be sufficient and necessary for circadian rhythms in olfactory responses. Given recent evidence for daily rhythms of pheromone responses in the antenna of the hawkmoth Manduca sexta, we examined whether a peripheral PER-based circadian clock might be present in this species. Several different cell types in the moth antenna were recognized by monoclonal antibodies against Manduca sexta PER. In addition to PER-like staining of pheromone-sensitive olfactory receptor neurons and supporting cells, immunoreactivity was detected in beaded branches contacting the pheromone-sensitive sensilla. The nuclei of apparently all sensory receptor neurons, of sensilla supporting cells, of epithelial cells, and of antennal nerve glial cells were PER-immunoreactive. Expression of per mRNA in antennae was confirmed by the polymerase chain reaction, which showed stronger expression at Zeitgeber-time 15 compared with Zeitgeber-time 3. This evidence for the expression of per gene products suggests that the antenna of the hawkmoth contains endogenous circadian clocks.