Brain-specific aldolase C amino acid sequence (greater than 75% of the coding region) was determined for the first time. Two cDNA clones, pAM1 and pAM2, were identified, from a mouse brain library, by using human aldolase B cDNA as a probe. The larger one, pAM2, identified as a cDNA for aldolase C, has been completely sequenced and covers the 5'-untranslated region of the mRNA and the codons for amino acids 1-227 of the protein. The sequence indicates that aldolase C is more akin to aldolase A than to aldolase B. A cDNA library from mouse muscle was also screened, allowing the identification of clones pAM3 and pAM4, which contain cDNAs for aldolase A. The sequence obtained from pAM3 covers 70% of the coding sequence (amino acids 99-355) from the -COOH part of the protein. The cDNAs for the three aldolases, A, B and C, have been hybridized to RNA from various rat tissues. The results confirm the tissue specificity of the expression of the mRNA for the different isoenzymes and support the hypothesis that aldolase C expression, as aldolase A and B, is regulated at the transcriptional level or, in any case, via mRNA concentration.