Molecular analyses of gene expression during sea star spermatogenesis

We have investigated actin gene expression during the annual spermatogenic cycle of Pisaster ochraceus by Northern blot analyses of testes RNAs pooled from defined spermatogenic stages. Specific probes for cytoplasmic (Cy) and muscle (M) actin gene products detect 2.3 and 2.1 kb transcripts, respectively. In addition, actin-coding sequence probes detect a third, much larger (3.5 kb) transcript designated FAT. Preliminary sequence analyses of two cDNAs representing portions of the FAT transcript show over 90% homology to Pisaster Cy actin at the amino acid level but only 80% nucleotide identity. The expression patterns of these three transcripts, plus two spermiogenic indicator transcripts (H3 histone and beta-tubulin), were determined over the cycle. The Cy transcript is seen at all stages but is ten- to 100-fold higher early in the cycle when mitotic activity predominates. The M transcript appears at the onset of gonadal growth and is maintained at constant levels through spermatogenesis consistent with the expansion of the muscular sheath surrounding the testes. The FAT, H3 histone, and beta-tubulin transcripts reach their highest levels in ripe testes when spermiogenic activity is maximal. The homology of the FAT transcript to actin, and its pattern of expression, suggest the hypothesis that this transcript may encode acrosomal actin.