We have cloned the full-length cDNA encoding the rat homolog of type I neurofibromin isoform, a protein product of a gene linked to neurofibromatosis type 1. Rat type I neurofibromin isoform is composed of 2,820 amino acid residues and shares about 98.5% amino acid identity with the human counterpart. By S1 nuclease mapping analysis of the alternatively spliced neurofibromin mRNAs in adult rat tissues, we showed that type I isoform mRNA was predominantly expressed in the brain, pituitary, and testis, while type II mRNA, carrying a 63-nucleotide insertion in the region coding for the domain related to GTPase-activating protein, was predominantly expressed in most other tissues, such as heart, kidney, and ovary. Furthermore, type II mRNA is predominant in the testis at age 1 week, while type I mRNA becomes predominant at 3 weeks and is subsequently expressed at higher levels, as seen in the adult testis. In contrast, type I mRNA is the predominant form in the brain throughout the postnatal development. Thus, the relative expression levels of type I and type II mRNAs may be specific to the tissues and to the developmental stage of certain tissues.