A long-term culture system for olfactory explants with intrinsically fluorescent cell populations

As a prerequisite for exploring the mechanisms which lead to the formation and maintenance of the precise wiring patterns in the olfactory system, organotypic cultures of olfactory tissue from transgenic mice expressing green fluorescent protein (GFP) under control of the olfactory marker protein promotor have been established. Tissue specimen from embryonic stage 14 were explanted and kept in culture for >1 week. Within the explants, numerous GFP-fluorescent olfactory sensory neurons assembled in an epithelial-like manner during this period. Under optimized culture conditions, strongly GFP-positive axons extended from these explants, fasciculated and formed bundles. When co-cultured with explants from the olfactory bulb, distinct axon populations were attracted by the target tissue; the fluorescent axon bundles invaded the bulbular explants and formed conglomerates at distinct spots. Explants from transgenic mice expressing GFP under control of a given olfactory receptor gene (mOR37A) also comprised labeled neurons that extended intensely fluorescent axonal processes, which all seemed to grow in a common fascicle. The results demonstrate that GFP-labeled olfactory sensory neurons differentiate in the established organotypic cultures, which thus appear to be a useful tool to monitor and to manipulate the processes underlying the axonal wiring between the olfactory epithelium and bulb.