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A genetically encoded sensor with improved fluorescence intensity for opioid detection at cellular resolution.
Kayla E Kroning1,2, Mingcheng Li1,2, D Isabel Petrescu1,3
1Life Sciences Institute, University of Michigan, Ann Arbor, Michigan, USA. wenjwang@umich.edu.
Chemical Communications (Cambridge, England)
|September 24, 2021
Summary
Researchers developed M-SPOTIT2, a brighter opioid sensor for detecting opioids at cellular resolution. This tool enhances understanding of opioid signaling in pain, reward, and respiration pathways.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- The mu-opioid receptor (MOR) is crucial for regulating pain, reward, and respiratory pathways.
- Detecting opioids at the cellular level is essential for understanding MOR's diverse roles.
Purpose of the Study:
- To engineer a more sensitive and brighter opioid sensor for cellular detection.
- To improve the tools available for studying opioid signaling.
Main Methods:
- Engineered M-SPOTIT2 by modifying a circularly permuted green fluorescent protein with specific amino acids (YNSH).
- Assessed sensor brightness in HEK293T cell culture and primary neuronal cultures.
Main Results:
- M-SPOTIT2 demonstrated an 11-fold increase in brightness compared to the previous M-SPOTIT1.1 in cell culture.
- M-SPOTIT2 showed a 2.7-fold increase in brightness in neuronal cultures.
Conclusions:
- M-SPOTIT2 is a significantly improved opioid sensor with enhanced brightness.
- This sensor has potential applications in drug screening and in vivo imaging of opioid signaling in animal tissues.
- M-SPOTIT2 can advance research into addiction, respiratory control, and pain modulation mechanisms.
