R34 BRAIN Initiative. Optical interrogation of neural circuits in Manduca
Project Summary / Abstract
This BRAIN Circuits Planning Project will help to bridge the gap between well-established brain studies
on the fruit fly, Drosophila, and complex behaviors exhibited more generally by other species. The
immediate goal is to adapt and apply optogenetic technologies to identify brain circuits responsible for
specific behaviors in the Tobacco Hawkmoth, Manduca sexta. In the long term, this will help to identify
fundamental principles underlying brain function by comparing Manduca brain circuits with those
underlying similar behaviors in Drosophila and other species. Manduca is an ideal organism for making
these comparisons. For more than 50 years Manduca has been an important experimental species for
understanding how nervous systems develop and how sensory signals in the environment are used to
control behavior. It is a large insect that is easily raised in the laboratory and its behavior, anatomy, and
physiology are known in detail. Furthermore, Manduca and Drosophila are superficially similar in their
anatomy and development, making many comparisons relatively straightforward, but the two species
also have distinct behavior and CNS organization. The experiments detailed in this project will
demonstrate that optogenetic components can be expressed in Manduca and used to monitor and
manipulate larval neural circuits and behavior. It will introduce new methods and improve on existing
techniques of Manduca gene targeting and transgenesis. This will include testing promoters to drive
expression of transgenes in central neurons, nociceptors and mechanosensors, validating their ability
to activate or inhibit neuronal activity, and assessing effects on behavior.
These experiments will prepare the groundwork for a BCP R01 project to identify the neural circuits
that underlie nociceptive behaviors in Manduca larvae. In Drosophila, nociception involves multiple
parallel circuits and the integration of different sensory modalities that are not fully understood. By
comparing nociceptive pathways in these species, it is expected any common organizing principles for
processing noxious or damaging stimuli will be identified. This will also inform studies of nociception
and pain perception carried out in other species including vertebrates. In addition to enabling future
work on nociception and locomotion, the tools developed here will be used by other researchers to
study different behaviors and life stages, including flight control, object tracking, navigation, and
multimodal integration.