NMDA receptor-dependent regulation of miRNA expression and association with Argonaute during LTP in vivo.
microRNAs (miRNAs) are major regulators of protein synthesis in the brain. A major goal is to identify changes in miRNA expression underlying protein synthesis-dependent forms of synaptic plasticity such as long-term potentiation (LTP). Previous analyses focused on changes in miRNA levels in total lysate samples. Here, we asked whether changes in total miRNA accurately reflect changes in the amount of miRNA bound to Argonaute protein within the miRNA-induced silencing complex (miRISC). Ago2 immunoprecipitation was used to isolate RISC-associated miRNAs following high-frequency stimulation (HFS)-induced LTP in the dentate gyrus of anesthetized rats. Using locked-nucleic acid-based PCR cards for high-throughput screening and independent validation by quantitative TaqMan RT-PCR, we identified differential regulation of Ago2-associated and total miRNA expression. The ratio of Ago2/total miRNA expression was regulated bidirectionally in a miRNA-specific manner and was largely dependent on N-methyl-D-aspartate receptor (NMDA) activation during LTP induction. The present results identify miRNA association with Ago2 as a potential control point in activity-dependent synaptic plasticity in the adult brain. Finally, novel computational analysis for targets of the Ago2-associated miRNAs identifies 21 pathways that are enriched and differentially targeted by the miRNAs including axon guidance, mTOR, MAPK, Ras, and LTP.
Department of Biomedicine and K.G. Jebsen Centre for Research on Neuropsychiatric Disorders, University of Bergen Bergen, Norway.
Balagopal
B
Pai
Department of Biomedicine and K.G. Jebsen Centre for Research on Neuropsychiatric Disorders, University of Bergen Bergen, Norway.
Taweeporn
T
Siripornmongcolchai
Department of Biomedicine and K.G. Jebsen Centre for Research on Neuropsychiatric Disorders, University of Bergen Bergen, Norway.
Birgitte
B
Berentsen
Department of Biomedicine and K.G. Jebsen Centre for Research on Neuropsychiatric Disorders, University of Bergen Bergen, Norway.
Ashraf
A
Pakzad
Department of Biomedicine and K.G. Jebsen Centre for Research on Neuropsychiatric Disorders, University of Bergen Bergen, Norway.
Christel
C
Vieuille
Department of Psychosocial Science, University of Bergen Bergen, Norway.
Ståle
S
Pallesen
Institute for Adaptive and Neural Computation, School of Informatics, University of Edinburgh Edinburgh, UK.
Maciej
M
Pajak
Institute for Adaptive and Neural Computation, School of Informatics, University of Edinburgh Edinburgh, UK ; Biomathematics and Statistics Scotland JCMB, Edinburgh, UK.
T Ian
TI
Simpson
Institute for Adaptive and Neural Computation, School of Informatics, University of Edinburgh Edinburgh, UK.
J Douglas
JD
Armstrong
Department of Biomedicine and K.G. Jebsen Centre for Research on Neuropsychiatric Disorders, University of Bergen Bergen, Norway.
Karin
K
Wibrand
Department of Biomedicine and K.G. Jebsen Centre for Research on Neuropsychiatric Disorders, University of Bergen Bergen, Norway.
Clive R
CR
Bramham
Argonaute; RNA-induced silencing complex; gene expression; hippocampus; microRNA; microRNA target prediction; protein synthesis; synaptic plasticity