College of Charleston
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My overall research interest is centered on understanding the diversity in protein composition within the flight muscles of insects. The aim for this project is focused on changes related to life cycle transition in the honeybee. All living creatures have the ability to change phenotypically (adapt), at least partially, in response to the demands of their environment (e.g. change in fur color between seasons in brown/white fox), as well as in response to physical demands (marathon versus sprint runners) and during progression through their life cycle. Among all insects the honeybee Apis mellifera is well suited for the study of plastic phenotypic adaptations. Honeybees are social insects, and within the hive many individuals start their life as worker nurses involved with building the honeycombs, feeding the larvae, tending the queen, and cleaning the hive. As such, nurse bees rarely need to fly and only limited distances for short times. Some of the nurse bees will transition later in life and become foragers whose job is to collect nectar; as such foragers need to be excellent flyers. The nurse-forager transition necessitates major changes in behaviors, metabolism, as well as flight muscle efficiency. Analysis of the troponinT protein, which is involved in calcium coupling within the myofilaments, indicates changes in protein isoform expression and relative amount. However differences in troponinT are not sufficient to explain the overall increase in forager’s flight performance and changes in isoform expression for other myofilament components are predicted. I generated RNA sequence data from RNA extracted from flight muscle of nurses and foragers. A differential gene expression analysis, as well as a qualitative and quantitative evaluation of splice variants distribution is being performed with a focus on proteins involved in myofilament structure. Besides the proteins of the myofilaments themselves, we expect to find variations in transcription and splicing factors, as well as for proteins involved in metabolism.
Honeybees were collected and assigned a preliminary classification as either nurse or forager subcaste based on behavior. Subsequently, bees weighing less than 0.7g were certified as foragers. The flight muscles were dissected from frozen bees and RNA extracted by standard Trizol protocol. Two replicates from each subcaste were processed for RNA sequencing.
Developmental Biology, Molecular Biology lectures and laboratories.
