Transfection of Rat primary hippocampal Neurons using Magnetofectamine Kit (CombiMag and L2000)

Rat primary hippocampal neurons were transfected after 11 DIV with 1µg plasmid using Lipofectamine 2000 and CombiMag, two transfection reagents associated in the Magnetofectamine Kit

This paper demonstrated the high efficiency of Magnetofectamine from OZ Biosciences to transfect primary hippocampal neurons. Numerous neuron types can efficiently be transfected using Magnetofection technology.

article reference: J Biol Chem. 2013 Feb 13.

ERK and GSK3β regulate gephyrin postsynaptic aggregation and GABAergic synaptic function in a calpain-dependent mechanism.
Tyagarajan SK, Ghosh H, Yevenes GE, Imanishi SY, Zeilhofer HU, Gerrits B, Fritschy JM.

Abstract

Molecular mechanisms of plasticity at GABAergic synapses are currently poorly understood. To identify signaling cascades that converge onto GABAergic postsynaptic density proteins we performed MS analysis using gephyrin isolated from rat brain and identified multiple novel phosphorylation and acetylation residues on gephyrin. Here, we report the characterization of one of these phospho-residues, S268, which when de-phosphorylated leads to the formation of larger postsynaptic scaffolds. Using a combination of mutagenesis, pharmacological treatment, and biochemical assays, we identify ERK as the kinase phosphorylating S268, and describe a functional interaction between residues S268 and S270. We further demonstrate that alterations in gephyrin clustering via ERK modulation are reflected by amplitude and frequency changes in miniature GABAergic postsynaptic currents. We unravel novel mechanisms for activity- and ERK-dependent calpain action on gephyrin, which are likely relevant in the context of cellular signaling affecting GABAergic transmission and homeostatic synaptic plasticity in pathology.

CombiMag from OZ Biosciences is a very efficient transfection reagent based on Magnetofection™ technology.