Efficient #transfection of #LNA (Locked Nucleic Acid) in #Primary #cortical and #hippocampal #neurons using #NeuroMag magnetofection reagent

Inhibition of the Schizophrenia-Associated MicroRNA miR-137 Disrupts Nrg1a Neurodevelopmental Signal Transduction

Kristen Therese Thomas et al. Cell Reports, 2017.

Efficient transfection of LNA (Locked Nucleic Acid) in Primary cortical and hippocampal neurons using NeuroMag magnetofection reagent

SUMMARY

Genomic studies have repeatedly associated variants

in the gene encoding the microRNA miR-137 

with increased schizophrenia risk. Bioinformatic predictions 

suggest that miR-137 regulates schizophrenia-

associated signaling pathways critical to 

neural development, but these predictions remain 

largely unvalidated. In the present study, we demonstrate 

that miR-137 regulates neuronal levels of 

p55g, PTEN, Akt2, GSK3b, mTOR, and rictor. All are 

key proteins within the PI3K-Akt-mTOR pathway 

and act downstream of neuregulin (Nrg)/ErbB and 

BDNF signaling. Inhibition of miR-137 ablates 

Nrg1a-induced increases in dendritic protein synthesis, 

phosphorylated S6, AMPA receptor subunits, 

and outgrowth. Inhibition of miR-137 also blocks 

mTORC1-dependent responses to BDNF, including 

increased mRNA translation and dendritic 

outgrowth, while leaving mTORC1-independent S6 

phosphorylation intact. We conclude that miR-137

regulates neuronal responses to Nrg1a and BDNF 

through convergent mechanisms, which might 

contribute to schizophrenia risk by altering neural 

development.

Transfections

For western blot experiments using LNA inhibitors, neurons were transfected on DIV11 or DIV12 with 75 nM LNA inhibitor (final concentration) by magnetofection using Neuromag (Oz Biosciences, NM50200) per manufacturer’s instructions, and neurons were lysed after 3 days (on DIV14 or DIV15, respectively).