Isolation of 10 Differentially Expressed cDNAs in Differentiated Neuro2a Cells Induced Through Controlled Expression of the GD3 Synthase Gene


Address correspondence and reprint requests to Dr. S. Tsuji at Department of Molecular Glycobiology, Frontier Research Program, Institute of Physical and Chemical Research, Wako, Saitama 351-0198, Japan.


Abstract: Recently, we showed that transfection of GD3 synthase cDNA into Neuro2a cells, a mouse neuroblastoma cell line, causes cell differentiation with neurite sprouting. In a search for the genes involved in this ganglioside-induced Neuro2a differentiation, we used a tetracycline-regulated GD3 synthase cDNA expression system combined with differential display PCRs to identify mRNAs that were differentially expressed at four representative time points during the process. We report here the identification of 10 mRNAs that are expressed highly at the Neuro2a differentiated stage. These cDNAs were named GDAP1–GDAP10 for (ganglioside-induced differentiation-associated protein) cDNAs. It is interesting that in retinoic acid-induced neural differentiated mouse embryonic carcinoma P19 cells, GDAP mRNA expression levels were also up-regulated (except that of GDAP3), ranging from three to > 10 times compared with nondifferentiated P19 cells. All the GDAP genes (except that of GDAP3) were developmentally regulated. The GDAP1, 2, 6, 8, and 10 mRNAs were expressed highly in the adult mouse brain, whereas all the other GDAP mRNAs were expressed in most tissues. Our results suggested that these GDAP genes might be involved in the signal transduction pathway that is triggered through the expression of a single sialyltransferase gene to induce neurite-like differentiation of Neuro2a cells.

Abbreviations used:

embryonic day


glyceraldehyde-3-phosphate dehydrogenase


ganglioside-induced differentiation-associated protein


postnatal day


retinoic acid


rapid amplification of cDNA ends


sodium dodecyl sulfate


salin–sodium citrate