Cytoplasmic localization during storage and translation

During spermatogenesis in mammals, the transcripts of transition protein 1 (TP 1) and protamine 1 (Prm 1) are under translational regulation. Following their transcription in round spermatids, the mRNAs for TP 1 and Prm 1 are stored in the cytoplasm from 3-7 days before being translated towards the end of spermatogenesis. To test the hypothesis that the inactivation or activation of transcripts during spermiogenesis could be mediated by mRNA compartmentalization in the cytoplasm of spermatids, light and electron microscopy were used to localize, by in situ hybridization, the cellular and subcellular sites of stored and translated mRNAs for these two testisspecific transcripts. During early spermiogenesis (before step 7) nuclear transcripts of both TP 1 and Prm 1 were seen. After step 7 the TP 1 and Prm 1 mRNAs were only detected in the cytoplasm. Throughout spermiogenesis the cytoplasmic mRNAs were not localized to any membrane-bound organelles such as the endoplasmic reticulum or mitochondria or to non-membrane-bound structures such as the chromatoid body. These studies demonstrate that the translational arrest of the TP 1 and Prm 1 mRNAs is not primarily controlled by compartmentalized storage in the cytoplasm of spermatids. Moreover, when translation of these mRNAs occurs in elongated spermatids, the mRNAs are present throughout the cytoplasm.