Primary microRNA precursor transcripts are localized at post-synaptic densities in adult mouse forebrain.
In a previous study, we reported that microRNA (miRNA) precursors are expressed in synaptic fractions within adult mouse forebrain, where they are enriched at post-synaptic densities (PSDs). However, because that study employed qRT-PCR primers that recognize the hairpin region, it was not able to distinguish between primary microRNA gene transcripts (pri-miRs) and small hairpin precursors (pre-miRs). Here, using primer sets that selectively measure regions upstream, downstream and flanking the hairpin, we demonstrate that pri-miRs are present in synaptic fractions (enriched several-fold relative to total tissue homogenate) and are especially enriched in isolated PSDs. Drosha and DGCR8 proteins are also expressed in synaptic fractions and PSDs, and are tightly associated with pri-miRs as assessed by coimmunoprecipitation under stringent conditions. Pri-miRs, drosha, and DGCR8 are highly enriched in fractions that contain mRNA transport particles, and cytosolic drosha is associated with kinesin heavy chain; these findings suggest that pri-miRs are transported to synaptic regions in a manner similar to mRNAs. This study supports the notion that miRNA biogenesis occurs locally near synapses in a regulated fashion.
Department of Psychiatry and Psychiatric Institute, University of Illinois at Chicago, Chicago, IL 60612, USA.
Giovanni
G
Lugli
John
J
Larson
Michael P
MP
Demars
Neil R
NR
Smalheiser
D000818:Animals; D047468:Immunoprecipitation; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000648:ultrastructure*; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D057907:Post-Synaptic Density / Q000378:metabolism*; D016548:Prosencephalon / Q000166:cytology*; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D016601:RNA-Binding Proteins; D043244:Ribonuclease III / Q000235:genetics / Q000378:metabolism; D013574:Synaptosomes / Q000378:metabolism
C474044:Dgcr8 protein, mouse; D035683:MicroRNAs; D011506:Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C513400:Drosha protein, mouse; D043244:Ribonuclease III; D010751:Phosphopyruvate Hydratase