Co-transfection of plasmid DNA into primary amygdal neurons using NeuroMag

Primary rat amygdal neurons were co-transfected with plasmid DNA using NeuroMag magnetic nanoparticles. 

This paper shows the high efficiency of NeuroMag from OZ Biosciences to transfect any kind of primary neurons such as maygdal neurons.

article reference: http://dx.doi.org/10.1016/j.psyneuen.2014.04.017

Functional effects of polymorphisms on glucocorticoid receptor modulation of human anxiogenic substance-P gene promoter activity in primary amygdala neurones

Colin W. Hay, Lynne Shanley, Scott Davidson, Philip Cowie, Marissa Lear, Peter McGuffin, Gernot Riedel, Iain J. McEwan, Alasdair MacKenzie.

 

Abstract

Expression or introduction the neuropeptide substance-P (SP; encoded by the TAC1 gene in humans and Tac1 in rodents) in the amygdala induces anxiety related behaviour in rodents. In addition, pharmacological antagonism of the main receptor of SP in humans; NK1, is anxiolytic. In the current study we show that the Tac1 locus is up-regulated in primary rat amygdala neurones in response to activation of the glucocorticoid receptor (GR); a classic component of the stress response. Using a combination of bioinformatics, electrophoretic mobility shift assays (EMSA) and reporter plasmid magnetofection into rat primary amygdala neurones we identified a highly conserved GR response sequence (2GR) in the human TAC1 promoter that binds GR in response to dexamethasone (Dex) or forskolin. We also identified a second GR binding site in the human promoter that was polymorphic and whose T-allele is only found in Japanese and Chinese populations. We present evidence that the T-allele of SNPGR increases the activity of the TAC1 promoter through de-sequestration or de-repression of 2GR. The identification of Dex/forskolin response elements in the TAC1 promoter in amygdala neurones suggests a possible link in the chain of molecular events connecting GR activation and anxiety. In addition, the discovery of a SNP which can alter this response may have implications for our understanding of the role of regulatory variation in susceptibility to stress in specific populations.

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. It can also be used on ES cell derived motor neurons.