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Updated: Jun 14, 2026
Deacetylation Assays to Unravel the Interplay between Sirtuins (SIRT2) and Specific Protein-substrates
Published on: February 27, 2016
Sirtuin 2 Regulates Dorsal Hippocampal Actin Polymerization and Microtubule Acetylation-Dependent EB3 Activation to
Jian-Dong Long1, Song-Yu Yao2, Wei-Wei Wu1
1Chinese Academy of Sciences Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China.
Background:
Aversive memories formed during drug withdrawal can potentiate compulsive drug-seeking behavior. Dynamic rearrangement of the actin cytoskeleton in dendritic spines is crucial for aversive memory formation. We previously discovered that actin polymerization in the dorsal hippocampus plays a key role in this process, but the molecular mechanisms governing this reorganization remain unclear.
Methods:
Conditioned morphine withdrawal-induced conditioned place aversion behavior in male Sprague Dawley rats was developed to evaluate aversive memory associated with opioid withdrawal. Contextual fear conditioning and appetitive conditioning paradigms were used to assess the behavioral specificity of the identified mechanisms. Western blots and dendritic spine imaging detected actin rearrangement and spine density. Co-immunoprecipitation, genetic manipulation, and pharmacological approaches were used to investigate sirtuin 2 (Sirt 2) and its downstream signaling.
Results:
We demonstrate that Sirt 2, an NAD+-dependent deacetylase, critically regulates dorsal hippocampal actin polymerization and opioid withdrawal-associated aversive memory through modification of microtubules (MTs). Sirt 2-induced tubulin deacetylation triggers EB3 dephosphorylation by promoting translocation of Ca2+-dependent phosphatase calcineurin to MTs. This enhances EB3-drebrin interaction, driving actin remodeling through the p140Cap/Src/cortactin/cofilin pathway. Furthermore, pharmacological inhibition of Sirt 2 also attenuated contextual fear conditioning and showed a trend toward reducing appetitive conditioning, indicating a shared role across multiple memory processes.
Conclusions:
The current study elucidates the molecular mechanisms by which Sirt 2 regulates dorsal hippocampal actin dynamics and drives opioid withdrawal memory and associative learning. Our findings reveal a broader role for Sirt 2 in multiple forms of associative memory, offering potential therapeutic insights for withdrawal-induced compulsive behaviors and other memory-related disorders.
