Differential mRNA expression of phospholipase D (PLD) isozymes during cAMP-induced differentiation in C6 glioma cells

GTP gamma S-dependent phospholipase D (PLD) activity time-dependently increased during differentiation of rat C6 glioma cells to astrocytic phenotypes induced by dibutyryl cyclic AMP (dbcAMP)/theophylline. The changes in PLD mRNA level were examined by reverse transcriptase-polymerase chain reaction (RT-PCR) method using the degenerate primers designed based on two conserved amino acid sequences in PLDs of human and yeast. The amplified three DNA fragments (tentatively termed as rPLDa, b, and c) contained the conserved regions present in PLDs of various organisms. RT-PCR using non-degenerate primers showed that rPLDa mRNA increased within 12h following treatment with dbcAMP, reaching a broad plateau and then returned to the initial level at 48h. In contrast, the level of rPLDb mRNA showed a concurrent decrease. rPLDc decreased in a time-dependent manner. These results suggest that the expression of PLD mRNAs are differentially regulated during differentiation in C6 glioma cells.