Large-scale investigation of the olfactory receptor space using a microfluidic microwell array
Open Access
- 10 February 2010
- journal article
- research article
- Published by Royal Society of Chemistry (RSC) in Lab on a Chip
- Vol. 10 (9), 1120-1127
- https://doi.org/10.1039/b920585c
Abstract
The mammalian olfactory system is able to discriminate among tens of thousands of odorant molecules. In mice, each odorant is sensed by a small subset of the approximately 1000 odorant receptor (OR) types, with one ORgene expressed by each olfactory sensory neuron (OSN). However, the sum of the large repertoire of OR–OSN types and difficulties with heterologous expression have made it almost impossible to analyze odorant-responsiveness across all OR–OSN types. We have developed a microfluidic approach that allowed us to screen over 20 000 single cells at once in microwells. By using calcium imaging, we were able to detect and analyze odorant responses of about 2900 OSNs simultaneously. Importantly, this technique allows for both the detection of rare responding OSNs as well as the identification of OSN populations broadly responsive to odorants of unrelated structures. This technique is generally applicable for screening large numbers of single cells and should help to characterize rare cell behaviors in fields such as toxicology, pharmacology, and cancer research.Keywords
This publication has 53 references indexed in Scilit:
- Odor Coding by a Mammalian Receptor RepertoireScience Signaling, 2009
- A multi-purpose microfluidic perfusion system with combinatorial choice of inputs, mixtures, gradient patterns, and flow ratesLab on a Chip, 2008
- Dual functions of mammalian olfactory sensory neurons as odor detectors and mechanical sensorsNature Neuroscience, 2007
- Fast Adaptation in Mouse Olfactory Sensory Neurons Does Not Require the Activity of PhosphodiesteraseThe Journal of general physiology, 2006
- Pheromone Detection in Male Mice Depends on Signaling through the Type 3 Adenylyl Cyclase in the Main Olfactory EpitheliumJournal of Neuroscience, 2006
- RTP Family Members Induce Functional Expression of Mammalian Odorant ReceptorsCell, 2004
- Tools for anti‐inflammatory drug design: In vitro models of leukocyte migrationMedicinal Research Reviews, 2004
- Preserved anterograde and retrograde memory of rapidly acquired olfactory discrminations after neurotoxic hippocampal lesionsHippocampus, 2003
- A novel multigene family may encode odorant receptors: A molecular basis for odor recognitionCell, 1991
- The maternal pheromone of the rat: Testing some assumptions underlying a hypothesisPhysiology & Behavior, 1983