Stage-dependent levels of specific mRNA transcripts in Sertoli cells

Abstract

Localization and stage-dependent levels of transferrin and sulfated glycoprotein-2 (SGP-2) mRNAs were examined in rat testes by in situ and soluble hybridization of mRNA with a single-stranded RNA probe prepared with the SP65 vector. Biotinylated RNA probes were identified in testicular tissue by using a biotinylated glucose oxidase-avidin system followed by a treatment with an appropriate electron carrier and a tetrazolium salt. This procedure demonstrated that the anatomical site of transferrin and SGP-2 gene expression was the Sertoli cells. Tritium-labeled RNA probes were visualized by radioautography. Negative and positive controls as well as in situ hybridization in Sertoli and myoid cells in culture indicated again that the cytoplasm of Sertoli cells was the anatomical site of transferrin and SGP-2 expression. Quantitative radioautography revealed cyclic variations in the level of both transferrin and SGP-2 mRNAs. The level of transferrin mRNA was relatively high from Stage I to Stage VIII. At Stage IX, the level decreased acutely and remained low in Stage X. The level of transcripts increased dramatically at Stage XIII, remaining high until Stage XIV. In the case of SGP-2 mRNAs, levels of transcripts were similar in most stages except at Stages VII and VIII, where higher levels were observed. These data were substantiated by similar results obtained by solution hybridization of both recombinant cRNAs with mRNAs from selected seminiferous tubules staged by transillumination. Thus, our results demonstrated a stage-specific regulation of transferrin and SGP-2 mRNA levels in Sertoli cells.