Expression of mRNAs encoding for two different olfactory receptors in a subset of olfactory receptor neurons.
Evidence has accumulated to support a model for odorant detection in which individual olfactory receptor neurons (ORNs) express one of a large family of G protein-coupled receptor proteins that are activated by a small number of closely related volatile chemicals. However, the issue of whether an individual ORN expresses one or multiple types of receptor proteins has yet to be definitively addressed. Physiological data indicate that some individual ORNs can be activated by odorants differing substantially in structure and/or perceived quality, suggesting multiple receptors or one nonspecific receptor per cell. In contrast, molecular biological studies favor a scheme with a single, fairly selective receptor per cell. The present studies directly assessed whether individual rat ORNs can express multiple receptors using single-cell PCR techniques with degenerate primers designed to amplify a wide variety of receptor sequences. We found that whereas only a single OR sequence was obtained from most ORNs examined, one ORN produced two distinct receptor sequences that represented different receptor gene families. Double-label in situ hybridization studies indicated that a subset of ORNs co-express two distinct receptor mRNAs. A laminar segregation analysis of the cell nuclei of ORNs labeled with the two OR mRNA probes showed that for one probe, the histogram of the distribution of the cell nuclei along the depth of the epithelium was bimodal, with one peak overlapping the (unimodal) histogram for the other probe. These results are consistent with co-expression of two OR mRNAs in a population of single ORNs.
Rawson
N E
NE
Monell Chemical Senses Center, and. Department of Pharmacology, University of Pennsylvania, Philadelphia, Pennsylvania, USA. rawson@monell.org
Eberwine
J
J
Dotson
R
R
Jackson
J
J
Ulrich
P
P
Restrepo
D
D
D000595:Amino Acid Sequence; D000818:Animals; D002469:Cell Separation; D015870:Gene Expression; D017403:In Situ Hybridization; D008297:Male; D008969:Molecular Sequence Data; D018034:Olfactory Receptor Neurons / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis*; D051381:Rats; D017207:Rats, Sprague-Dawley; D018035:Receptors, Odorant / Q000737:chemistry / Q000235:genetics*
D012333:RNA, Messenger; D018035:Receptors, Odorant