Adenosine deaminase-related genes: Molecular identification, tissue expression pattern and truncated alternative splice isoform in adult zebrafish ( Danio rerio)

Abstract

Adenosine deaminase (ADA) is responsible for cleaving the neuromodulator adenosine to inosine. Two members of ADA subfamilies, known as ADA1 and ADA2, were described and evidence demonstrated another similar protein group named ADAL (adenosine deaminase “like”). Although the identification of ADA members seems to be consistent, the expression profile of ADA1, ADA2 and ADAL genes in zebrafish has not yet been reported. The aim of the present study was to map the expression pattern of ADA-related genes in various tissues of adult zebrafish ( Danio rerio). An extensive search on zebrafish genome followed by a phylogenetic analysis confirmed the presence of distinct ADA-related genes ( ADA1, ADAL and two orthologous genes of ADA2). Specific primers for each ADA member were designed, optimized semi-quantitative RT-PCR experiments were conducted and the relative amount of transcripts was determined. The tissue samples (brain, gills, heart, liver, skeletal muscle and kidney) were collected and the expression of ADA1, ADAL and ADA2 genes was characterized. ADA1 had a similar expression pattern, whereas ADAL was less expressed in the heart. The highest relative amount of ADA2-1 transcripts was observed in the brain, liver and gills and it was less expressed in the heart. RT-PCR assays revealed that the other ADA2 form ( ADA2-2) was expressed ubiquitously and at comparable levels in zebrafish tissues. The strategy adopted also allowed the identification of an ADA2-1 truncated alternative splice isoform ( ADA2-1/T), which was expressed at different intensities. These findings demonstrated the existence of different ADA-related genes, their distinct expression pattern and a truncated ADA2-1 isoform, which suggest a high degree of complexity in zebrafish adenosinergic system.