Disruption of persistent, stress-associated memories is relevant for treating posttraumatic stress disorder (PTSD) and related syndromes, which develop in a subset of individuals following a traumatic event. We previously developed a stress-enhanced fear learning (SEFL) paradigm in inbred mice that produces PTSD-like characteristics in a subset of mice, including persistently enhanced memory and heightened cFos in the basolateral amygdala complex (BLC) with retrieval of the remote (30-day-old) stress memory. Here, the contribution of BLC microRNAs (miRNAs) to stress-enhanced memory was investigated because of the molecular complexity they achieve through their ability to regulate multiple targets simultaneously. We performed small-RNA sequencing (smRNA-Seq) and quantitative proteomics on BLC tissue collected from mice 1 month after SEFL and identified persistently changed microRNAs, including mir-135b-5p, and proteins associated with PTSD-like heightened fear expression. Viral-mediated overexpression of mir-135b-5p in the BLC of stress-resilient animals enhanced remote fear memory expression and promoted spontaneous renewal 14 days after extinction. Conversely, inhibition of BLC mir-135b-5p in stress-susceptible animals had the opposite effect, promoting a resilient-like phenotype. mir-135b-5p is highly conserved across mammals and was detected in post mortem human amygdala, as well as human serum samples. The mir-135b passenger strand, mir-135b-3p, was significantly elevated in serum from PTSD military veterans, relative to combat-exposed control subjects. Thus, miR-135b-5p may be an important therapeutic target for dampening persistent, stress-enhanced memory and its passenger strand a potential biomarker for responsivity to a mir-135-based therapeutic.
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We thank the Scripps Florida Genomics Core for sequencing services, Nripesh Prasad at the Genomic Services Lab at Hudson Alpha for sequencing services and data analysis, Adrian Reich and the Bioinformatics Core for data analysis, the Mouse Behavior core and Alicia Brantley for assistance and behavioral equipment, all members of the Miller/Rumbaugh Labs for their technical assistance and thoughtful discussions. This work was funded by grants from the National Institute of Mental Health MH105400 and MH105400-02 (Diversity Supplement) (CM), National Institute of Neurological Disorders and Stroke NS096833 (CM), National Institute on Drug Abuse DA041469 (SS) and the Brain and Behavior Foundation-NARSAD Young Investigator Award (SS). This research project was supported in part by the Viral Vector Core of the Emory Neuroscience National Institute of Neurological Disorders and Stroke Core Facilities grant, P30NS055077. LdN and smRNA-Seq experiments in human serum were funded by a VIDI award number 91718336 from the Netherlands Scientific Organization (BR) and the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie grant agreement No. 707362 (LDN).
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Sillivan, S.E., Jamieson, S., de Nijs, L. et al. MicroRNA regulation of persistent stress-enhanced memory. Mol Psychiatry 25, 965–976 (2020). https://doi.org/10.1038/s41380-019-0432-2
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