Transient receptor potential (TRP) channels are a superfamily of cation channels that regulate processes as diverse as olfaction, thermal perception, pain, synaptic transmission and development. They can be activated by both environmental stimuli (e.g. chemicals, light, temperature, pressure) and intracellular stimuli (e.g. calcium and G protein coupled receptors). More than 50 different TRP channels have been identified in organisms ranging from yeasts and protists to mammals. The experiments in this project will extend the study of TRP channels into one of the most primitive metazoans with a nervous system, the cnidarian Hydra. A characteristic of all the Cnidaria is the possession of stinging cells called cnidocytes, which in Hydra are responsible for prey capture, adhesion and locomotion. Although significant progress has been made in understanding development and regeneration in Hydra, the molecular mechanisms that control Hydra behaviors such as its feeding response and cnidocyte release are as of yet poorly understood. An examination of the Hydra genome revealed the presence of multiple candidate Hydra TRP channel genes. Whole mount in situ hybridization will be used to localize TRP channel expression to specific Hydra body regions. TRP channel function will be examined by testing known chemical TRP channel activators and inhibitors to determine if they affect Hydra feeding behavior or cnidocyte release, and if they affect the ability of Hydra to regenerate or to form buds during asexual reproduction. In addition, RNA interference will be used to silence the expression of specific Hydra TRP channels. The resulting TRP(-) hydra will then be tested in the functional assays to determine the effect of TRP channel gene silencing. The study of Hydra TRP channels could answer fundamental questions about the signal transduction systems involved in Hydra behaviors and development. A comparison of the primitive TRP channels in Hydra with the more recently evolved TRP channels of higher organisms will provide a useful mechanism for both structure-function analysis and for studying evolutionary relationships.
BROADER IMPACTS Queensborough Community College (QCC) is an urban community college with a student population comprised of nearly equal numbers of Asians, Caucasians, Latinos and African Americans. Many of QCC's students come from low-income families, and are the first in their family to attend college. One of the goals of this project is to afford QCC undergraduates the opportunity to do research in a scientific laboratory. In the course of their research, students will learn Hydra culture methods, basic molecular biology techniques and DNA/protein sequence analysis. As the research in this project spans the spectrum from DNA sequence to development and behavior, students will learn how to make connections between events at the molecular level and those occurring at the level of the entire organism. In addition, since the TRP channel gene family is found in both very simple organisms such as yeast and in complex organisms like mammals, students will need to think in evolutionary terms when analyzing TRP channel function. Firsthand experience gained by working in a laboratory allows students to connect the theoretical concepts they learn in the classroom with real-world experiences. This will enhance their understanding of scientific concepts, and will give them the opportunity to apply their knowledge to their experiments in the laboratory. Student participation in research increases retention and graduation rates, a particularly important benefit at a community college like QCC, and undergraduates who are exposed to research early during their college careers are more likely to choose STEM majors and to remain in STEM careers.