MAL mRNA is induced during the differentiation of human embryonal carcinoma cells into neurons and is also localised within specific regions of the human brain

Abstract

We have found that the MAL gene, which encodes a membrane proteolipid expressed during the late stages of T-lymphocyte maturation, is also activated during neuronal differentiation of NTERA2 human embryonal carcinoma cells following induction with retinoic acid. An RT-PCR fragment with a sequence identical to MAL was found amongst 30 cloned DNA fragments corresponding to genes putatively activated during NTERA2 differentiation and isolated using a differential display PCR screen. PCR and Northern blot analysis with a cloned MAL cDNA as a probe confirmed that MAL is not expressed by undifferentiated NTERA2 EC cells, but is expressed, predominantly as a 1.1 kb transcript, within 7 days of retinoic acid-induced differentiation and later in the post-mitotic neurons arising in such cultures. MAL was not expressed in the non-neuronal lineages induced by treatment of NTERA2 cells with the gratuitous inducer hexamethylene bisacetamide. Analysis of cDNA libraries constructed from EC cells, purified neurons and a sub-population of non-neuronal cells (ME311+), confirmed that expression of the MAL gene is activated in the neural lineage of NTERA2 differentiation. Using in situ hybridisation we found that MAL is expressed in the human CNS and especially in grey matter of the cerebral cortex, with less in the cerebellum and the amygdala and little or none in subcortical white matter. In contrast to reports concerning the expression pattern of a rat MAL homologue, MAL was expressed in the human brain predominantly in cell bodies which include neurons, correlating with in vitro data from the NTERA2 line.