Hippocampal neurons transfection with DNA and siRNA using the Magnetofectamine Kit (CombiMag and L2000)

Culture of primary mice hippocampal neurons were transfected at 12 DIV with DNA construct and/or siRNA 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 Neurosci. 2013 Oct 16;33(42):16828-45.

The Neural Cell Adhesion Molecule Promotes Maturation of the Presynaptic Endocytotic Machinery by Switching Synaptic Vesicle Recycling from Adaptor Protein 3 (AP-3)- to AP-2-Dependent Mechanisms.

Shetty A, Sytnyk V, Leshchyns'ka I, Puchkov D, Haucke V, Schachner M.

Abstract

Newly formed synapses undergo maturation during ontogenetic development via mechanisms that remain poorly understood. We show that maturation of the presynaptic endocytotic machinery in CNS neurons requires substitution of the adaptor protein 3 (AP-3) with AP-2 at the presynaptic plasma membrane. In mature synapses, AP-2 associates with the intracellular domain of the neural cell adhesion molecule (NCAM). NCAM promotes binding of AP-2 over binding of AP-3 to presynaptic membranes, thus favoring the substitution of AP-3 for AP-2 during formation of mature synapses. The presynaptic endocytotic machinery remains immature in adult NCAM-deficient (NCAM-/-) mice accumulating AP-3 instead of AP-2 and its partner protein AP180 in synaptic membranes and vesicles. NCAM deficiency or disruption of the NCAM/AP-2 complex in wild-type (NCAM+/+) neurons by overexpression of AP-2 binding-defective mutant NCAM interferes with efficient retrieval of the synaptic vesicle v-SNARE synaptobrevin 2. Abnormalities in synaptic vesicle endocytosis and recycling may thus contribute to neurological disorders associated with mutations in NCAM.

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