Immunofluorescent localization of a murine seminal vesicle proteinase inhibitor

The indirect immunofluorescent technique was used to localize a proteinase inhibitor isolated from murine seminal vesicles. The inhibitor was found in the lumen and in the apical epithelium of the seminal vesicle but not in the testes, epididymides, ductus deferens or Cowper's glands. It was also associated with the anterior acrosomal region of ejaculated sperm and sperm recovered from the female tract within 5 min of coitus. The inhibitor is removed from uterine sperm between 2 to 4 h postcoitus, however sperm recovered from the uterus 2 h postcoitus will rebind inhibitor. The inhibitor is not normally associated with epididymal or ductus sperm although these sperm will bind purified inhibitor in vitro.     

The role of the seminal vesicles, coagulating glands and prostate glands on the fertility and fecundity of mice.

Female mice were allowed to mate with males which had been sham-operated (Group 1); had seminal vesicles, coagulating glands and ventral and dorsal prostate glands removed (Group 2); had the seminal vesicles removed (Group 3); had the coagulating glands removed (Group 4) or had the ventral and dorsal prostate gland removed (Group 5). The pregnancy rate was normal in Groups 1 and 4, severely reduced in Groups 2 and 3 and less so in Group 5. Litter size was reduced in Groups 2 and 3 but not in Group 5. It is suggested that the seminal vesicles and possibly the prostate glands are important in the production of young in mice.     

Effects of seminal vesicle removal on fertility and uterine sperm motility in the house mouse

The relative significance of the accessory glands of the male reproductive tract in fertility is unclear. To clarify the role of the seminal vesicles, fertility and uterine sperm motility were determined before and after removal of seminal vesicles in the house mouse. After removal of seminal vesicles, the pregnancy rate (number of females pregnant/number of females X 100) was reduced and the time to birth was increased, while the average litter size was not changed. Fertilization, determined by examining the oocytes 30 h after mating, was highly variable after matings with males whose seminal vesicles were removed; in some cases none of the oocytes were fertilized. The motility of sperm recovered from the uterus 1 h after matings with males before and after seminal vesicle removal and sham operations was analyzed using a videomicrographic system. The motility of uterine sperm was less progressive with more lateral displacement of the head about the trajectory and a less linear trajectory after removal of the seminal vesicles. Sham-operated animals showed no consistent changes in motility of uterine sperm. The changes in sperm motility could contribute to the reduction in fertilization since sperm motility is necessary for transport in the female reproductive tract and interaction with the oocytes.     

Ultrastructure and function of the seminal vesicle of Bittacidae (Insecta: Mecoptera)

The fine structure of the seminal vesicle and reproductive accessory glands was investigated in Bittacidae of Mecoptera using light and transmission electron microscopy. The male reproductive system of Bittacidae mainly consists of a pair of testes, a pair of vasa deferentia, and an ejaculatory sac. The vas deferens is greatly expanded for its middle and medio-posterior parts to form a well-developed seminal vesicle. The seminal vesicle is composed of layers of developed muscles and a mono-layered epithelium surrounding the small central lumen. The epithelium is rich in rough endoplasmic reticulum and mitochondria, and secretes vesicles and granules into the central lumen by merocrine mechanisms. A pair of elongate mesodermal accessory glands opens into the lateral side of the seminal vesicles. The accessory glands are similar to the seminal vesicle in structure, also consisting of layers of muscle fibres and a mono-layered elongated epithelium, the cells of which contain numerous cisterns of rough endoplasmic reticulum and mitochondria, and a few Golgi complexes. The epithelial cells of accessory glands extrude secretions via apocrine and merocrine processes. The seminal vesicles mainly serve the function of secretion rather than temporarily storing spermatozoa. The sperm instead are temporarily stored in the epididymis, the greatly coiled distal portion of the vas deferens.     

The reproductive tract of the male spiny mouse (Acomys cahirinus) and coagulation studies with other species

The testes of the spiny mice showed asymmetry, the left being significantly heavier than the right (P = 0.025). Histological studies indicated that spermatozoa were first present in the testes of animals 35--45 days of age but the maturation of the accessory glands, especially the lateral prostates and coagulating glands, occurred later. The highest fructose concentration in the adult was in the lateral prostates (126.97 +/- 22.23 mg fructose/100 g, n = 5) and coagulating glands (99.38 +/- 17.65 mg fructose/100 g gland weight, n = 5). Coagulation tests of mixtures of extracts of seminal vesicles and coagulating glands from spiny mice and rats indicated that the vesiculase of the spiny mouse was active on rat substrates and vice versa. Cross-reactions of extracts of house mouse (Mus musculus), hamster (Mesocricetus auratus), gerbil (Meriones unguiculatus), and guinea-pig (Cavia porcellus) seminal vesicles (substrate) and coagulating glands (vesiculase) with those of rats and spiny mice showed that although the substrates of rat and spiny mouse were readily coagulated by vesiculase from all the other species, rat and spiny mouse vesiculase were not equally active on substrates of the other species.     

Decreased fructose concentrations in coagulating glands of prepuberal rats treated with testosterone and LH

Doses of testosterone propionate from 2.5 to 320 microgram and doses of LH from 2 to 360 microgram given over 1--3 days generally decreased fructose/body weight ratios in the coagulating glands of late prepuberal rats. The ratios of testes, seminal vesicles, coagulating glands and ventral prostates to body weight were increased after different treatment regimes with testosterone propionate. These changes in the variables measured could be detected by computer analysis in spite of the rapid growth rates of organs of rats of this age. LH increased the weights of only the seminal vesicles and coagulating glands, and then, only at the highest doses given.     

A propos des vésicules séminales: le point de vue du biologiste

Progress in assisted procreation techniques has led to decreased interest in seminal vesicle investigations. However, recent progress in comparative anatomy and physiology emphasize the important role played by the seminal vesicles in fertilization. Seminal vesicle secretions play a role in the mechanics of coitus, on the behaviour of spermatozoa and the female genital tract and they also protect against infections. They consequently facilitatein vivo fertilization and the andrologist should pay particular attention to the seminal vesicles to ensure physiological procreation under good conditions.     

Decapacitation and recapacitation of rabbit spermatozoa treated with membrane vesicles from seminal plasma

The article presents 2 experiments to determine whether a dense vesicle fraction concentration (Fr.1) of seminal plasma membrane vesicles could decapacitate rabbit spermatozoa and whether spermatozoa decapacitated by this method could recapacitate. It was found that 9k to 1 mg. concentration of Fr.1/ml. impaired spermatozoic fertility in dose 2 hours after ovulation. Epididymal fluid, free of sperm cells, impaired the fertility of spermatozoa capacitated in utero. This result can be explained by the high content of Fr.1-type vesicles in the epididymis. Fertilization was achieved in 19 of 28 (68%) does with the deposit of decapacitated spermatozoa obtained 7 hours after mating in the uterus about 6 hours before ovulation. This is not significantly lower than the fertilization rate obtained with untreated spermatozoa (80%). Spermatozoa obtained 13 hours after mating showed a poor fertilization capacity when deposited 6 hours before ovulation.     

Elucidation of the protein composition of mouse seminal vesicle fluid

The seminal vesicles are male accessory sex glands that contribute the major portion of the seminal plasma in which mammalian spermatozoa are bathed during ejaculation. In addition to conveying sperm through the ejaculatory duct, seminal vesicle secretions support sperm survival after ejaculation, and influence the female reproductive tract to promote receptivity to pregnancy. Analysis of seminal vesicle fluid (SVF) composition by proteomics has proven challenging, due to its highly biased protein signature with a small subset of dominant proteins and the difficulty of solubilizing this viscous fluid. As such, publicly available proteomic datasets identify only 85 SVF proteins in total. To address this limitation, we report a new preparative methodology involving sequential solubilization of mouse SVF in guanidine hydrochloride, acetone precipitation, and analysis by label-free mass spectrometry. Using this strategy, we identified 126 SVF proteins, including 83 previously undetected in SVF. Members of the seminal vesicle secretory protein family were the most abundant, accounting for 79% of all peptide spectrum matches. Functional analysis identified inflammation and formation of the vaginal plug as the two most prominent biological processes. Other notable processes included modulation of sperm function and regulation of the female reproductive tract immune environment. Together, these findings provide a robust methodological framework for future SVF studies and identify novel proteins with potential to influence both male and female reproductive physiology.     

Seminal vesicle secretion of African catfish, its composition, its behaviour in water and saline solutions and its influence on gamete fertilizability

The seminal vesicle secretion (SVS) of the African catfish, Clarias gariepinus, was investigated by analytical and experimental methods. SVS consists mainly of proteins and glycoproteins which are responsible for its viscous and sticky nature. The secretion contains also high activities of acid phosphatase, alkaline phosphatase, and proteases. These catabolic enzymes do not have functions in autolysis or liquefaction of SVS but are considered to eliminate aging spermatozoa from the proximal portions of seminal vesicle and from the spermatic duct. SVS of the African catfish is unstable in the environment relevant for natural spawning. When SVS was mixed with water, seminal plasma or different types of saline solutions its protein coagulated forming fibrous or granular particles of variable size within a few seconds. Pure SVS completely inhibited the motility as the sticky secretion hindered spermatozoa in free swimming. SVS had also a negative effect on sperm fertility, egg fertility, and sperm egg contact, as the fertilization was drastically suppressed in the presence of SVS. Basing on our analytical and experimental results we exclude that SVS has functions in stabilizing the viability of spermatozoa stored in the spermatic ducts or is an energy resource of spermatozoa. It also does not improve or stabilize the fertilization process and has no functions in adhering the eggs to substrates or in covering the eggs for mechanical protection or antibacterial defense. A function of SVS in the male and female communication during the prenuptial spawning behaviour is discussed.     

The effect of vesiculectomy on seminal characteristics in the stallion.

Successful unilateral extirpation of an inflamed seminal vesicle in a stallion led to systematic trials of the influence of a reduction and absence of the secretion of this gland upon semen characteristics. Operations were performed by the method described for the bull. The volume of ejaculates dropped and sperm concentration per ml increased in each of 2 stallions from which the seminal vesicles had been uni- or bi-laterally removed. Total sperm number and motility remained uninfluenced, but the percentage of eosin-stained spermatozoa increased in the unilaterally operated stallion and the percentage of abnormal spermatozoa increased significantly in both. Concentration of citric acid per ml and per ejaculate was significantly reduced after bilateral vesiculectomy. Ergothioneine concentration per ml increased in the unilaterally and in the bilaterally operated stallion.     

The relationship between the morphology of cell organelles and kinetics of the secretory process in male sex accessory glands of mice

Summary Two male sex accessory glands of the mouse, seminal vesicle and coagulating gland, were compared with the aim of relating differences in the morphology of organelles to the kinetics of the secretory process. The epithelial cells of the two glands were assessed by morphometric analysis, cytochemical staining, and electron-microscopic autoradiography after administration of a labeled amino acid. The rough endoplasmic reticulum of the seminal vesicle comprised narrow parallel cisternae, while that of the coagulating gland was greatly distended and occupied a much larger percentage of the cytoplasmic volume. Radioactively labeled products were secreted much more rapidly in the seminal vesicle than in the coagulating gland. The primary point of difference in kinetics of intracellular transport between the two glands was in exit of material from the rough endoplasmic reticulum. The more rapid drainage of the rough endoplasmic reticulum may be related to its relatively greater membrane surface density and lesser internal volume. In contrast, similarities in size and cytochemical staining in the Golgi apparatus of the two glands were accompanied by similar kinetics of intracellular transport of secretory protein through this organelle.     

Localization and characterization of glutathione peroxidase (GPx) in boar accessory sex glands, seminal plasma, and spermatozoa and activity of GPx in boar semen

Boar ejaculate owes its characteristic large volume mainly to accessory sex gland (ASG) secretions. These are main contributors to the protective functions of seminal plasma, especially against oxidative damage. Numerous antioxidants have been detected in ASG secretions, and, respectively, in seminal plasma. However, as regards one key antioxidant protector -- the Se-dependent enzyme glutathione peroxidase (GPx) -- there is no agreement yet among researchers as to its presence in boar seminal plasma. Nevertheless, the beneficial effect of dietary Se supplementation on male fertility has been widely recognized. The aim of the present study was to investigate the localization and characterization of GPx in boar ASGs, seminal plasma, and spermatozoa, as well as to evaluate GPx activity in boar semen. Immunohistochemical assays demonstrated GPx presence in the epithelial cells, vacuole membranes, and vascular endothelium of boar seminal vesicle, prostate and bulbourethral glands. Western blot analysis demonstrated the presence of a monomer form of GPx with MW 20 kDa in lysates from seminal vesicle, prostate, bulbourethral glands, and spermatozoa, but not in seminal plasma. Surprisingly, peroxidase activity detected in seminal plasma from normal ejaculates was nearly three times as high as in spermatozoa. Our findings confirmed the presence of immunoreactive GPx in the boar reproductive tract, while further investigation is still warranted to uncover the exact protein forms involved and their function.     

Semen-coagulating protein, SVS2, in mouse seminal plasma controls sperm fertility

Mammalian seminal plasma is known to contain a decapacitation factor(s) that prevents capacitation and thus, the fertility of sperm. This phenomenon has been observed in experiments conducted in vitro that assessed the inhibition of epididymal sperm fertility by seminal plasma or by the purified decapacitation factor. However, the phenomenon of decapacitation has not yet been characterized in vivo. In the present study, we demonstrate that seminal vesicle protein secretion 2 (SVS2), which is a 40-kDa basic protein and a major component of the copulatory plug, enters the uterus and interacts with ejaculated sperm heads after copulation. The SVS2-binding region of sperm changed from the postacrosomal region to the equatorial segment, while the sperm migrated through the uterus and finally disappeared in the oviduct. Furthermore, SVS2 reduced the fertility of epididymal sperm. The sperm treated with SVS2 decreased the percentage of fertilized oocytes from 60% to 10%. The capacitation state was assessed by protein tyrosine phosphorylation and the comprehensiveness of the acrosome reaction. SVS2 functioned to maintain sperm in the uncapacitated state and to reverse capacitated sperm to the uncapacitated state. We found that the fertility of ejaculated sperm is associated with SVS2 distribution in the female reproductive tract. These results indicate that SVS2 functions as a decapacitation factor for mouse sperm.     

Effects of seminal vesicle fluid components on sperm motility in the house mouse

Fluid obtained by stripping dissected seminal vesicles was mixed with phosphate-buffered saline and the soluble proteins were separated by gel filtration on BioRad P150 into 4 fractions. Fractions were collected and concentrated using an Amicon ultrafiltration system using YM2 membranes with a molecular weight cut-off of 1000. Epididymal sperm suspensions were incubated in medium containing one of the 4 fractions or 1 mg BSA/ml, or no added protein. After incubation for 2 h the motility of the spermatozoa in each suspension was assessed by a videomicrographic procedure. Two aspects of motility, velocity and the shape of the swimming path, were monitored. The results indicate that the seminal vesicles produce at least three factors that influence sperm motility. Fraction 3 (Mr 12,000-24,000) was detrimental to motility; after incubation for 2 h almost all the spermatozoa were immotile. Fractions 2 (Mr 25,000-40,000) and 4 (Mr 7000-12,000) both influenced the shape of the swimming path: spermatozoa incubated in Fraction 2 had straighter trajectories while those incubated in Fraction 4 showed more progressive paths with less side-to-side movement of the head about the path. These effects of factors from the seminal vesicle fluid on sperm motility may influence the way in which the spermatozoa move in the female reproductive tract and could help to explain why removal of the seminal vesicles reduces fertility in the mouse.     

Carcinogenesis in accessory sex organs of rats induced by 3,2′-dimethyl-4-aminobiphenyl: response to androgen deprivation

 It is generally accepted that early human prostate cancers reveal higher androgen dependency than do advanced ones. In the present study, we examined whether the animal model of prostate cancer has already lost androgen dependency at the early stages of carcinogenesis. At experimental week 46, androgen deprivation was induced in rats and the incidences of atypical hyperplasia and cancer were examined in the ventral, dorsolateral prostate, coagulating glands, and seminal vesicles. Androgen deprivation significantly lowered the incidence of atypical hyperplasia in all four organs. As for the incidence of cancer, no significant differences were observed in the coagulating glands and seminal vesicles. Regarding atypical hyperplasia, androgen deprivation significantly decreased the proliferative cell nuclear antigen labeling index in the coagulating gland and seminal vesicles. The presence of cancer was also decreased in the coagulating gland but not in the seminal vesicles. With control group specimens, more intense staining of androgen receptor was observed in atypical hyperplasias than in cancers. Compared with the atypical hyperplasias, the cancers revealed low androgen dependency at the early stages of carcinogenesis. The cancers in the seminal vesicles also revealed higher androgen independency than did those in the coagulating gland. Accepted: 6 May 1997     

SERPINE2, a serine protease inhibitor extensively expressed in adult male mouse reproductive tissues, may serve as a murine sperm decapacitation factor

SERPINE2, one of the potent serine protease inhibitors that modulates the activity of plasminogen activator and thrombin, is implicated in many biological processes. In the present study, we purified SERPINE2 from mouse seminal vesicle secretion (SVS), using liquid chromatography and identified it by liquid chromatography/tandem mass spectrometry, and it showed potent inhibitory activity against the urokinase-type plasminogen activator. SERPINE2 was expressed predominantly in seminal vesicles among murine male reproductive tissues. It was immunolocalized to the SVS and mucosal epithelium of the seminal vesicle, epididymis, coagulating gland, and vas deferens. In the testes, SERPINE2 was immunostained in spermatogonia, spermatocytes, spermatids, Leydig cells, and spermatozoa. SERPINE2 was also detected on the acrosomal cap of testicular and epididymal sperm and was suggested to be an intrinsic sperm surface protein. The purified SERPINE2 protein could bind to epididymal sperm. A prominent amount of SERPINE2 was detected on ejaculated and oviductal spermatozoa. Nevertheless, SERPINE2 was detected predominantly on uncapacitated sperm, indicating that SERPINE2 is lost before initiation of the capacitation process. Moreover, SERPINE2 could inhibit in vitro bovine serum albumin-induced sperm capacitation and prevent sperm binding to the egg, thus blocking fertilization. It acts through preventing cholesterol efflux, one of the initiation events of capacitation, from the sperm. These findings suggest that the SERPINE2 protein may play a role as a sperm decapacitation factor.     

The phospholipid-binding protein of the reproductive tract of the bull — Effect on the removal of spermatozoal cytoplasm droplets in other species and influence of antibodies on its reactivity

The phospholipid-binding protein (PBP) isolated from bull seminal vesicle fluid removed cytoplasm droplets not only from bull, but also from ram, boar and rabbit epididymal spermatozoa. However, the presence of a protein cross-reacting with anti-PBP antisera was demonstrated by immunofluorescent staining in ram seminal vesicles and ampullae. In contrast to PBP from bull, the ram PBP-like protein did not lyse bull or ram erythrocytes. Rabbit antiserum against PBP only negligibly reduced the ability of PBP to remove cytoplasm droplets from bull epididymal spermatozoa, but it inhibited the haemolytic effect of the protein.     

A major secretory protein from rat seminal vesicle binds ejaculated spermatozoa

The rat seminal vesicle produces large amounts of a protein-rich fluid that greatly contributes to semen volume. RSV IV, a protein abundantly secreted from this gland, binds in vitro to rat epididymal spermatozoa. However, there is no evidence that this protein may have an in vivo role as a sperm-coating antigen. We report in this paper that high-molecular-weight RSV IV immunologically related proteins can be detected on ejaculated spermatozoa, but not on epididymal spermatozoa. After incubation of purified RSV IV with ejaculated spermatozoa in freshly recovered semen or with epididymal spermatozoa in a medium containing the coagulating gland secretion, sperm-bound proteins with analogous properties were detected. These results support the hypothesis that RSV IV is modified at ejaculation to an high-molecular-weight, sperm-coating antigen.     

Ultrastructure and histochemistry of the male reproductive system of the genus Callinectes Stimpson, 1860 (Brachyura: Portunidae)

We described the ultrastructure and histochemistry of the reproductive system of five Callinectes species, and evaluate the seasonal variation in weight of the reproductive system and hepatopancreas by comparing annual changes of somatic indices. The somatic indices changed little throughout the year. In Callinectes, spermatogenesis occurs inside the lobular testes and, within each lobule, the cells are at the same developmental stage. Spermatogenesis and spermiogenesis follow the same development pattern in all Callinectes studied. Mature spermatozoa are released into the seminiferous ducts through the collecting ducts. Cells of the vas deferens are secretory as evidenced by rough endoplasmic reticulum, Golgi complex, and secretory vesicles that produce the seminal fluid. The anterior vas deferens shows two portions: proximal and distal. In proximal portion (AVDp), spermatozoa are clustered and embedded in an electron-dense, basophilic glycoproteinaceous secretion Type I. In the distal portion (AVDd), the spermatophore wall is formed by incorporation of a less electron-dense glycoproteinaceous secretion Type II. The secretion Type I change to an acid polysaccharide-rich matrix that separates the spermatophores from each other. The median vas deferens (MVD) stores the spermatophores and produces the granular glycoproteinaceous seminal fluid. The posterior vas deferens (PVD) has few spermatophores. Its epithelium has many mitochondria and the PVD seminal fluid changes into a liquid and homogeneous glycoprotein. Many outpocketings in the PVD and MVD help to increase the fluid production. Overall, the reproductive pattern of Callinectes is similar to other species that produce sperm plugs. The secretions of AVD, MVD, and PVD are responsible for the polymerization that forms the solid, waxy plug in the seminal receptacle. The traits identified here are common to all Portunidae species studied so far.