Tissue distribution
Northern blot analyses of the Acdp gene family were carried out using membranes purchased from Origene. A total of 12 mouse tissues were included in the study (Fig. 1). Due to sequence homologies between each Acdp member within the conserved domain, probes for Northern bolts were PCR fragments from the last exon and the 3' untranslated region sequences. The mouse Acdp messages showed almost the same tissue distributions as the human ACDP genes. Acdp1 message is highly expressed in the brain, while kidney and testis also showed low levels of expression. Acdp2 showed higher expressions in the brain, kidney and liver. However, the Acdp2 transcript was not present in the skeleton muscle and skin, and it showed very low levels of expression in the rest of tissues. Acdp3 and Acdp4 showed different levels of expression in all tissues tested; the highest expressions for Acdp3 were observed in the brain, kidney, liver and heart, and the highest expressions for Acdp4 were observed in the kidney, small intestine and testis. The expression levels for Acdp3 and 4 in skeleton muscle were barely detectable; however, β-actin showed normal expression suggesting that the results were not a consequence of bad RNA quality (data not shown). The ubiquitous expression pattern may be taken as another indication of the functional importance of Acdp proteins in fundamental biological processes in addition to the sequence conservation in evolutionarily divergent species.
Figure 1  Northern blot analyses of the Acdp gene family. S. muscle represents skeletal muscle, Sm. Int. represents small intestine. Multiple Choice Northern Blot filters were purchased from Origene.