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Highly sensitive in vivo detection of dynamic changes in enkephalins following acute stress in mice
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Highly sensitive in vivo detection of dynamic changes in enkephalins following acute stress in mice
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Journal Article
Highly sensitive in vivo detection of dynamic changes in enkephalins following acute stress in mice
2025
Overview
Enkephalins are opioid peptides that modulate analgesia, reward, and stress. In vivo detection of enkephalins remains difficult due to transient and low endogenous concentrations and inherent sequence similarity. To begin to address this, we previously developed a system combining in vivo optogenetics with microdialysis and a highly sensitive mass spectrometry-based assay to measure opioid peptide release in freely moving rodents (Al-Hasani et al., 2018, eLife). Here, we show improved detection resolution and stabilization of enkephalin detection, which allowed us to investigate enkephalin release during acute stress. We present an analytical method for real-time, simultaneous detection of Met- and Leu-enkephalin (Met-Enk and Leu-Enk) in the mouse nucleus accumbens shell (NAcSh) after acute stress. We confirm that acute stress activates enkephalinergic neurons in the NAcSh using fiber photometry and that this leads to the release of Met- and Leu-Enk. We also demonstrate the dynamics of Met- and Leu-Enk release as well as how they correlate to one another in the ventral NAc shell, which was previously difficult due to the use of approaches that relied on mRNA transcript levels rather than posttranslational products. This approach increases spatiotemporal resolution, optimizes the detection of Met-Enk through methionine oxidation, and provides novel insight into the relationship between Met- and Leu-Enk following stress.
Publisher
eLife Sciences Publications Ltd,eLife Sciences Publications, Ltd
Subject
/ Animals
/ Biochemistry and Chemical Biology
/ Brain
/ Enkephalin, Leucine - analysis
/ Enkephalin, Leucine - metabolism
/ Enkephalin, Methionine - analysis
/ Enkephalin, Methionine - metabolism
/ Genetics
/ in vivo
/ liquid chromatography-mass spectrometry
/ Male
/ Mice
/ Nucleus Accumbens - metabolism
/ Opioids
/ Optics
/ Peptides
/ stress
