Technical Reference #1624

1624.

Valproic Acid Induces Up- or Down- Regulation of Gene Expression Responsible for the Neuronal Excitation and Inhibition in Rat Cortical Nuerons Through its Epegenetic Actions Mamoru Fukuchi; Takuya Nii; Naoki Ishimaru; Aya Minamino; Daichi Hara; Ichiro Takasaki; Akiko Tabuchi; Masaaki Tsuda, University of Toyama, Neuroscience Research, 65(1624), (2009)

Abstract:
Valproic acid (VPA) a drug used to treat epilepsy and bipolar mood disorder inhibits histone deacetylase(HDAC) which is associated with the epigenetic regulation of gene expression. Using a microarray wecomprehensively examined which genes are aff

Keywords:
VPA; HSAC inhibitor; histone acetylation; BDFN; GABAa receptor; GAD

Materials & Methods:
2.5. Measurement of the intracellular Ca2+ concentration in neurons Cortical cells were seeded in a poly-L-lysine-coated glassbottom dish (MatTek Corporation Ashland MA). The change in [Ca2+]i in single cells was measured by the fura-2 technique with minor modification. The cells were incubated with 10 mM fura-2 acetoxymethylester (fura-2-AM Boehringer Mannheim GmbH Dojindo Kumamoto Japan) for 30 min at 37 8C in DMEM. The cells were then washed with balanced salt solution (BSS 150 mM NaCl 5 mM KCl 1.2 mM CaCl2 1.2 mM MgCl2 10 mM glucose and 25 mM HEPES pH 7.4) and mounted on an inverted fluorescence microscope (TE300 Nikon Tokyo Japan). Digital images of fura-2 fluorescence recorded by a high-sensitivity CCD camera (C2400- 87 Hamamatsu Photonics Shizuoka Japan) were analyzed by a digital image processor (Argus-20 Hamamatsu Photonics). The ratio of 510 nm emission fluorescence at 340 nm excitation to that at 380 nm excitation F (340/380) was used as an indicator of [Ca2+]i in single cells.

Microscopic Technique
Fluorescence Microscopy

Cell Type(s)
Neurons