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Chem. Senses 29: 179-187, 2004
© Oxford University Press 2004

Primary Culture of Lobster (Homarus americanus) Olfactory Sensory Neurons

Ruben Stepanyan, Bettye Hollins, S. Erin Brock and Timothy S. McClintock

Department of Physiology, University of Kentucky, Lexington, KY 40536-0298, USA

Correspondence to be sent to: Dr Timothy S. McClintock, Louis Boyarski Associate Professor of Physiology, Department of Physiology, University of Kentucky, Lexington, KY 40536-0298, USA. e-mail: mcclint{at}uky.edu

Lobster olfactory sensory neurons have contributed to a number of advances in our understanding of olfactory physiology. To facilitate further study of their function, we have developed conditions allowing primary culture of the olfactory sensory neurons in a defined medium. The most common cells in the culture were round cell bodies with diameters of 10–15 µm that often extended fine processes, features resembling olfactory sensory neurons. We discovered that acetylcholinesterase acted as a growth factor for these cells, improving their survival in culture. We also confirmed previous evidence from spiny lobsters that poly-D-lysine was a superior substrate for olfactory cells of this size and morphology. We then identified olfactory sensory neurons in the culture in two ways. Almost half the cells tested responded to application of a complex odorant with an inward current. An even more rigorous test was made possible by the development of an antiserum to OET-07, an ionotropic glutamate receptor homolog specifically expressed by Homarus americanus olfactory sensory neurons. It labeled a majority of the round cells in the culture, unequivocally identifying them as olfactory sensory neurons.

Key words: : acetylcholinesterase, crustacea, glutamate receptor, smell


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