Tissue and developmental distribution, dependence upon testicular factors and attachment to spermatozoa of GPX5, a murine epididymis-specific glutathione peroxidase.

Using immunohistochemistry and Western blotting analyses, we present a detailed study of the distribution of the glutathione peroxidase protein (GPX5) within the mouse epididymis. We have shown that the expression of the epididymis-specific protein is restricted to the caput and essentially localized to the apical cell border of the caput epithelium. Secretion of the protein was detected as early as the proximal segment of the caput and GPX5 was subsequently found in the lumen of corpus and cauda epididymis duct. Within the caput, Western blot analyses have shown that equivalent quantities of GPX5 protein were found in segments I, II, and III. During ontogenesis, GPX5 appeared at 20 days postnatal, before the completion of the morphological differentiation of the caput and concomitantly with the appearance of spermatozoa within the epididymis, in agreement with what was reported earlier regarding the transcription of its corresponding gene during epididymal ontogenesis (Faure et al., 1991). Hormonal privation by castration abolished the accumulation of the GPX5 protein confirming previous data obtained on GPX5 mRNA levels. Treatments such as testosterone replacement or hemicastration led to the restriction of the protein to the caput epithelium, suggesting that protein secretion partly depends both on the presence of testicular factors and on spermatozoa. Using electron microscopy, we have shown that the secreted protein binds to spermatozoa and is found predominantly on the sperm acrosomic region. Finally, we report here that the GPX5 protein can be detected in fluids recovered from the uterine horns of freshly mated female mice. These results suggest that GPX5 might play an important role in sperm maturation from the early events up to the onset of fertilization and therefore could potentially be used as a tool to monitor sperm quality.