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Using reverse-transcription polymerase chain reaction (RT-PCR), we identified three alternative splicing pathways of the ret proto-oncogene (proto-ret) which occur at intron 4. Although the majority of the primary transcripts were spliced to make mRNA encoding the protein product previously reported, two alternatively spliced transcripts containing either a 62 bp or a 69 bp insertion were also present. Intron 4 of proto-ret was cloned from human genomic DNA using a PCR method, and both the 62 bp and 69 bp sequences were localized in the intron. Furthermore, using a semi-quantitative RT-PCR method, we found that the expression of the two alternatively spliced transcripts varies in cell lines derived from different tissues. The relative amount of the transcripts containing the 62 bp insertion in these cell lines was: THP-1 > NB-39-nu = TT, whereas the relative amount of the transcripts containing the 69 bp insertion was: THP-1 > TT > NB-39-nu.
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