oa Evidence for a cell fate refinement mechanism in sensory neurons

The olfactory receptors (ORs), which are G-protein coupled receptors, number more than 1,000 and comprise the largest gene family in the mammalian genome. ORs are expressed both monogenically and monoallelically in olfactory sensory neurons (OSNs) and the mechanism that controls their regulation is largely unknown. ORs reside in constitutive heterochromatin and selection of one OR and from one allele occurs stochastically. To ensure singular monoallelic OR expression, a negative feedback mechanism is elicited by the first selected OR to suppress non-selected ORs in a given neuron. Here we describe results for mice with a 'monoclonal' nose that express one OR, M71,in 95% of all mature OSNs. The M71 transgene suppresses expression of endogenous ORs, and remaining endogenous expression is mostly restricted to immature neurons in the olfactory epithelium. We show that the endogenous OR repertoire are expressed prior to suppression by M71 transgene expression, contrary to current models. When we introduced a second transgene into M71 mice that expressed another OR in most mature OSNs, OSNs uncharacteristically expressed both of the ORs. We hypothesize that unresolved OR competition compromised the neuron's ability to express only one receptor. We further show that suppression of endogenous ORs by M71 is not reversible, and that M71 does not need to be continuously expressed for endogenous ORs to remain suppressed. In these experiments, we have engineered OSNs to express more than one OR in an OSN at a time, which is normally a low probability event. We believe that these experiments reveal the existence of a backup pathway that ensures only one OR will be expressed per neuron. It is possible that mammals have evolved similar mechanisms in other examples of stochastic, monoallelic gene expression, such as in random X-chromosome inactivation or lymphocyte receptor expression.