Synthesis, characterization and hormonal regulation of epididymal proteins during postnatal development of the mouse

Abstract Maturation of mammalian spermatozoa depends on their interactions with epididymal proteins. The incorporation of 35 S-methionine into these proteins was investigated by in vitro incubation of tissue minces from the mouse epididymis at different ages of postnatal development. The greatest amount of incorporation per wet weight of tissue was seen in 7 to 21-day-old mice. It decreased progressively during development while the rate of proteins released into the medium remained almost constant until the adult state. Separation of labeled proteins on sodium dodecyl sulphate polyacrylamide gels followed by fluorography showed that the great majority of secretory proteins synthesized in adult mouse epididymis could be recovered already from 7-day-old animals. Regional differences appeared at 21 days of age. These were marked by the secretion of proteins characteristic of the proximal (26, 25, 20, 19 kDa) and distal (44, 29 kDa) epididymis. Analysis of cytosol and luminal fluid proteins from prepubertal and adult epididymis revealed a number of proteins of the same mobility as those synthesized and secreted in vitro. Among the luminal proteins which showed variations during development and regional differences, four (29, 26, 20, 19 kDa) were characteristics of the epididymis and three (88, 34, 13 kDA) comigrated with testicular components. Castration or estrogen treatment of prepubertal mice for 4, 3 and 2 weeks inhibited or reduced the synthesis of the luminal proteins which appeared during postnatal development and/or presented regional differences. Testosterone replacement of castrated mice reversed this effect and induced the secretion of new proteins (37, 24 kDa). Combined administration of estrogen and testosterone to castrated or intact prepubertal mice reduced the androgen effect on protein synthesis. These results indicate that epididymal differentiation is characterized by the secretion of epididymal segment-specific and androgen-regulated proteins which appear early during development even before the sperm arrival in the epididymis.