We have isolated vesicular structures from mouse epididymal fluid, referred to as epididymosomes. Epididymosomes have a roughly spherical aspect and a bilayer membrane, and they are heterogeneous in size and content. They originate from the epididymal epithelium, notably from the caput region, and are emitted in the epididymal lumen by way of apocrine secretion. We characterized their membranous lipid profiles in caput and cauda epididymidal fluid samples and found that epididymosomes were particularly rich in sphingomyelin (SM) and arachidonic acid. The proportion of SM increased markedly during epididymal transit and represented half the total phospholipids in cauda epididymidal epididymosomes. The cholesterol:phospholipid ratio increased from 0.26 in the caput to 0.48 in the cauda epididymidis. Measures of epididymosomal membrane anisotropy revealed that epididymosomes became more rigid during epididymal transit, in agreement with their lipid composition. In addition, we have characterized the membrane lipid pattern of murine epididymal spermatozoa during their maturation. Here, we have shown that mouse epididymal spermatozoa were distinguished by high percentages of SM and polyunsaturated membranous fatty acids (PUFAs), principally represented by arachidonic, docosapentanoic, and docosahexanoic acids. Both SM and PUFA increased throughout the epididymal tract. In particular, we observed a threefold rise in the ratio of docosapentanoic acid. Epididymal spermatozoa had a constant cholesterol:phospholipid ratio (average, 0.30) during epididymal transit. These data suggest that in contrast with epididymosomes, spermatozoal membranes seem to become more fluid during epididymal maturation.

In mice, GPX5 is a secreted protein abundantly synthesized by the caput epididymidis. The protein is secreted as early as the initial segment of the caput and is found subsequently associated with the sperm plasma membrane in a sub-acrosomic localization. We show here that GPX5 is present in the caput and cauda epididymides lumens in three different locations: either free as a soluble protein in the caput epididymal fluid, weakly bound to caput sperm membranes, or, finally, associated to lipid-containing structures conferring to the protein a protective effect against proteolytic digestions. Within the cauda epididymidis, the amount of free GPX5 is low compared to the caput and the association with sperm membranes proved to be more solid. In both caput and cauda sperm samples, the association of GPX5 with the sperm membrane protects GPX5 from proteolytic cleavages. Protection against proteolytic digestions can be overcome by physical treatments of epididymal fluid and sperm samples such as ultrasounds or very acidic pH. These data suggest that complex phenomena and structures participate in the transfer and binding of the caput-secreted GPX5 protein to the sperm plasma membrane.

__~-_ ~y~ RESUME Des v~sicules de secr(~tions apocrines ont 6t(~ observees dans la lumiere de nombreux tissus du tractus g~nital m&le. Longtemps considerees comme des artefacts de fixation des tissus, elles sont maintenant reconnues comme des elements b part enti6re qui jouent des r61es physiologiques dans la maturation des gametes m~les. L'objet de cette revue est de presenter les diff6rentes structures vdsiculaires qui ont ete decrites chez les mammiferes tant sur les plans de leurs organisations, contenus et fonctions potentielles. ABSTRACT Apocrine secretions in the male genital tract: putative roles in sperm maturationHanae REJRAJI, Jo~l R. DREVET Apocrine secretory vesicles have been observed in the lumen of several ducts of the male genital tract, namely the epididymis and vas deferens. Initially considered to be tissue-fixation artefacts, they are now recognized as genuine elements, which might play physiological roles in terms of sperm maturation. The aim of this review is to present these vesicles observed in various mammalian species and to describe their respective organization, content and putative roles.

Un des aspects de la maturation posttesticulaire des spermatozoïdes chez les mammifères est l'évolution progressive de la composition lipidique de la membrane plasmique des gamètes mâles qui permet à ces derniers d'acquérir des capacités fusogènes particulières détermi-nantes pour la fécondation. Une part importante de cette modification du contenu lipidique de la membrane des gamètes mâles est assurée lors de la descente des spermatozoïdes dans le tubule épididymaire. Les mouvements lipidiques seront ensuite poursuivis dans les voies génitales femelles au cours d'un processus dynamique, la capacitation, étape préalable indispensable à la réaction acrosomique, et donc, à la fécondation. Cette revue fait l'état des lieux des connaissances acquises par divers groupes de recherche dans le premier volet épididymaire de la maturation lipidique des gamètes mâles dans différents modèles de mammifères.Abstract One aspect of mammalian post-testicular sperm maturation is the progressive change in their plasma membrane lipid composition. These modifications in lipids allow sperm cells to fuse with oocytes during fertilization. A significant share of these sperm lipid changes occurs during their descent through the epididymal tubule. It then continues within the female genital tract during the capacitation process, an essential prerequisite for acrosomic reaction and hence fertilization. This review presents what is known concerning the sperm plasma membrane lipid changes during epididymal maturation in various mammalian models. In the first section, after a brief presentation of the classic eukaryotic cell plasma membrane lipid organization, the emphasis is on the particularities of sperm plasma membrane lipids. The second section presents the different changes occurring in the three major classes of lipids (i.e. phospholipids, sterols and fatty acids) during the sperm's epididymal descent. The final section briefly describes the mechanisms by which these lipid changes might happen in the epididymal lumen environment. The role played by lipid-rich vesicles secreted by the epididymal epithelium via apocrine secretory processes is highlighted.