Receptor Array for Odor Discrimination via Cell Activation Measurement
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Solution Overview
Problem
Current methods lack a technique for constructing a system to measure the degree of activation of multiple types of olfactory receptors, which is necessary for creating a sensitive odor detection device that can discriminate various odors, as existing systems fail to express olfactory receptors at a sufficient level and maintain efficient intracellular signal transduction.
Innovation Solution
A receptor array with a substrate and hydrophobic coating film featuring through-holes, where nucleic acids coding for specific receptors are placed, and eukaryotic cells expressing G protein-coupled receptors are used to measure the activation degree by calculating the ratio of ion uptake before and after depolarization, allowing for comprehensive evaluation of receptor activation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional detection apparatus such as gas chromatography is used, then the device structure is simple, but the odor discrimination capability is insufficient
Solution Approach 1:
The invention copies the biological odor recognition mechanism of olfactory sensory neurons into an artificial detection system. By expressing olfactory receptors on the cell surface of cultured cells and measuring their activation responses, the system replicates the high discrimination capability of biological noses while maintaining a relatively simple in vitro assay format
Solution Approach 2:
The invention replaces the mechanical/physical separation and detection mechanism of gas chromatography with a biological recognition system. Instead of physically separating odor components and detecting them individually, the system uses olfactory receptors to biologically recognize and respond to odor molecules, achieving superior discrimination through biological specificity
2Measurement precision
If multiple types of olfactory receptors are constructed into an array, then the odor discrimination capability improves, but the measurement technique complexity increases
Solution Approach 1:
The invention uses a universal measurement approach that can detect the activation of multiple different olfactory receptor types using the same basic assay protocol. By employing a common signaling pathway (cAMP accumulation) that is shared across different olfactory receptors, the system can simultaneously measure the activation of numerous receptor types without requiring separate measurement techniques for each
Solution Approach 2:
The invention measures receptor activation by detecting changes in intracellular cAMP concentration, a biochemical parameter that changes in response to odorant binding. This parameter-based measurement approach allows for quantitative comparison of activation across multiple receptor types while maintaining a consistent and relatively simple measurement methodology
3Reliability
If olfactory receptors are expressed on cultured cells, then the signal transduction capability improves, but the expression level and signal transduction efficiency are insufficient
Solution Approach 1:
The invention introduces a signaling intermediary approach by measuring downstream effects (cAMP accumulation) rather than directly measuring receptor binding or activation. This intermediary measurement method amplifies the signal and makes it easier to detect, thereby improving the reliability of signal transduction measurements while maintaining accurate reflection of receptor activation
Solution Approach 2:
The invention employs preliminary optimization of cell culture conditions and receptor expression systems to ensure sufficient receptor expression levels before measurement. By pre-establishing optimal transfection protocols and cell culture conditions, the system achieves reliable signal transduction without requiring complex real-time adjustments during measurement
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
Enables the precise measurement of receptor activation degrees, facilitating the creation of a sensitive odor detection device capable of discriminating various odors by expressing multiple olfactory receptors on cells and calculating their activation levels.
Implementation Method 1
a hydrophobic coating film on the substrate, wherein the hydrophobic coating film has a plurality of through-holes
Implementation Method 2
Receptors cause signal transduction by action with an extracellular substance (ligand). Olfactory receptors are activated by stimulation of an odor molecule from the outside of cells, and Gaolf protein, a type of G protein, is then activated in cells
Implementation Method 3
Olfactory receptors are a type of 7-transmembrane receptor expressed in olfactory sensory neurons, and interact with G proteins in the intracellular region of olfactory receptors
Implementation Method 4
This Gaolf activates adenylate cyclase and accumulates cAMP in olfactory sensory neurons using ATP as a raw material
Implementation Method 5
Then cAMP is bound to Cyclic Nucleotide Gated ion-channel (CNG) on the cell membrane of an olfactory sensory neuron to open the channel, and calcium ion flows into cells through CNG
Implementation Method 6
consequently, the concentration of calcium ion in the cells increases
Data Source
AI summary
Method(s) for constructing a device for causing a plurality of receptors to be expressed in respectively different eukaryotic cells upon nucleic acids within through-holes, housing a plurality of eukaryotic cells within the through-holes of the device and causing the receptors to be expressed therein, comprehensively measuring the degree of activation of the plurality of receptors by a test substance, and evaluating the degree of activation as a specific numerical value. Provided is an array which includes a substrate and a hydrophobic coating film upon the substrate, the hydrophobic coating film having a plurality of through-holes, and a nucleic acid containing a gene which codes for a prescribed receptor being provided in the interior of each through-hole in a manner touching the substrate.