Responses to odors mapped in snail tentacle and brain by [14C]-2-deoxyglucose autoradiography

The method of 2-deoxyglucose (2-DG) autoradiography has been widely used to map functional neuronal systems in vertebrates, but in invertebrate species, where morphological dimensions favor its use, the applications have been minimal. This study uses [14C]-2-DG to map the olfactory system of a terrestrial snail, Achatina fulica. The olfactory organ in the snail's tentacles bears a striking resemblance to the vertebrate olfactory mucosa. There are also complex neural structures in the tentacle and brain that are devoted to subsequent processing. These facts make the molluscan olfactory system a suitable complement to the traditional vertebrate and insect models in olfaction. The experiments utilized intact snails in which one tentacle was exposed to a controlled odor environment while the contralateral tentacle was held in a retracted position. The dose of [14C]-2-DG (2 microCi/gm) was injected into the hemocele. Tissue processing involved freeze-substitution with acetone, dry sectioning, and the preparation of liquid film autoradiographs. Optical density measurements permitted quantitative comparisons between experimental conditions. The natural odors of conspecific snails and of carrots elicited significantly more uptake of 2-DG in the exposed tentacle than in the unexposed tentacle and, in the exposed tentacle, significantly more label over the axons of the primary sensory neurons than was elicited by exposure to clean air. Amyl acetate and octanol were less effective. A small number of superficially placed sensory neurons were labeled in all stimulus conditions, including clean air, and may represent the mechanosensors. Stimulus-dependent labeling in the brain was limited to the procerebrum and included both neuropilar and cellular parts. In contrast to vertebrate and insect olfactory systems, there was no evidence of spatial coding for odor quality.