DIV 15 primary rat hippocampal neurons transfection using NeuroMag

Primary hippocampal neurons from Rats at DIV 15 were transfected with a plasmid encoding for shRNA using NeuroMag reagent (Magnetofection). Experiments were performed 6 days after transfection.

This paper shows the high efficiency of NeuroMag transfection reagent from OZ Biosciences to transfect long term cultures (DIV 15) of primary hippocampal neurons with shRNA encoding plasmid and its lack of toxicity.

article reference: PLoS One. 2014 Mar 21;9(3):e92309. 

EphA4 Activation of c-Abl Mediates Synaptic Loss and LTP Blockade Caused by Amyloid-β Oligomers.

M Vargas L, Leal N, Estrada LD, González A, Serrano F, Araya K, Gysling K, Inestrosa NC, Pasquale EB, Alvarez AR.

Abstract

The early stages of Alzheimer's disease are characterised by impaired synaptic plasticity and synapse loss. Here, we show that amyloid-β oligomers (AβOs) activate the c-Abl kinase in dendritic spines of cultured hippocampal neurons and that c-Abl kinase activity is required for AβOs-induced synaptic loss. We also show that the EphA4 receptor tyrosine kinase is upstream of c-Abl activation by AβOs. EphA4 tyrosine phosphorylation (activation) is increased in cultured neurons and synaptoneurosomes exposed to AβOs, and in Alzheimer-transgenic mice brain. We do not detect c-Abl activation in EphA4-knockout neurons exposed to AβOs. More interestingly, we demonstrate EphA4/c-Abl activation is a key-signalling event that mediates the synaptic damage induced by AβOs. According to this results, the EphA4 antagonistic peptide KYL and c-Abl inhibitor STI prevented i) dendritic spine reduction, ii) the blocking of LTP induction and iii) neuronal apoptosis caused by AβOs. Moreover, EphA4-/- neurons or sh-EphA4-transfected neurons showed reduced synaptotoxicity by AβOs. Our results are consistent with EphA4 being a novel receptor that mediates synaptic damage induced by AβOs. EphA4/c-Abl signalling could be a relevant pathway involved in the early cognitive decline observed in Alzheimer's disease patients.

NeuroMag is the first dedicated Magnetofection ™ transfection reagent for neurons. It is perfect for primary neurons and can also be used with cell lines and glial cells. The transfection can be performed with neurons from 3 DIV to 21 DIV.