Hippocampal neurons were transfected at DIV 15 using 0.5 µg DNA and 1.75 µL of NeuroMag transfection reagent. Analysis was performed at DIV 20.
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 DNA.
article reference: Mol Cell Neurosci. 2013 Aug 8. pii: S1044-7431
Ezh1 and Ezh2 differentially regulate PSD-95 gene transcription in developing hippocampal neurons. Henriquez B,
Bustos FJ,
Aguilar R,
Becerra A,
Simon F,
Montecino M,
van Zundert B.
Abstract
Polycomb Repressive Complex 2 (PRC2)
mediates transcriptional silencing by catalyzing histone H3 lysine 27
trimethylation (H3K27me3), but its role in the maturation of postmitotic
mammalian neurons remains largely unknown. We report that the PRC2
paralogs
Ezh1 and
Ezh2 are
differentially expressed during hippocampal development. We show that depletion of
Ezh2 leads to increased expression of
PSD-95, a critical plasticity gene, and that reduced
PSD-95 gene transcription is correlated with enrichment of
Ezh2 at the
PSD-95 gene promoter; however, the H3K27me3 epigenetic mark is not present at the
PSD-95
gene promoter, likely due to the antagonizing effects of the H3S28P and
H3K27Ac marks and the activity of the H3K27 demethylases JMJD3 and UTX.
In contrast, increased
PSD-95 gene transcription is accompanied by the presence of
Ezh1 and elongation-engaged RNA Polymerase II complexes at the
PSD-95 gene promoter, while knock-down of
Ezh1 reduces
PSD-95 transcription. These results indicate that
Ezh1 and
Ezh2 have antagonistic roles in regulating
PSD-95 transcription.
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.
No comments:
Post a Comment