Identification and evaluation of NO-regulated genes by differential analysis of primary cDNA library expression (DAzLE)

Nitric oxide (NO) has numerous physiological roles in the cell. One of the actions of NO is gene regulation through protein modification and signal transduction. In neurons, NO can be produced from neuronal NO synthase, which is activated by calcium following N-methyl-D-aspartate (NMDA) receptor activation. Differential analysis of cDNA library expression (DAzLE) was used to identify differentially expressed genes by NO. Fundamentally, this technique combines differential hybridization to isolate genes whose expression is differentially regulated with microarray to analyze the expression of the isolated genes. The expression of genes identified by the DAzLE method is verified further by quantitative real-time polymerase chain reaction (RT-PCR) and/or Northern blot analysis. The high selectivity and sensitivity of this technique for detecting differentially expressed gene transcripts enable the investigation and identification of a panel of genes that are regulated by NO.