Deet technical fact sheet from the National Pesticide Information Center
The purpose of our project was to determine the impact of oxidative stress and the effects of exogenous antioxidants on neurite growth in neurons extracted from the X-organ/sinus gland of the fiddler crab Uca pugilator.
After removing eyestalks from anesthetized crabs, we microdissected the eyestalks and removed neural tissue from the X-organ, located directly opposite the opalescent-blue sinus gland. We digested the tissue fragments enzymatically using trypsin, separated the tissue to break loose individual neurons, and cultured the cells for a period of forty-eight hours.
Neural cells were cultured in a normal culture medium were used as controls. Some cells were cultured in a medium treated with 100% DEET insect repellent (at a 50% lethal dose), while others were cultured in a medium treated with 24% standardized Ginkgo Biloba extract (10X the human dose).
After forty-eight hours, the cell cultures were examined under a light microscope at 40X magnification and photographed using AxioVision software. Neurite area was established using Image J, and the sizes of the cell bodies were assayed.