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Obst, M., Nakano, H., Bourlat, S.J., Thorndyke, M.C., Telford, M.J., Nyengaard, J.R. and Funch, P. 2011. Spermatozoon ultrastructure of Xenoturbella bocki (Westblad 1949). - Acta Zoologica (Stockholm) : 109-115. Here, we report on the sperm ultrastructure of Xenoturbella bocki (Westblad 1949), which we studied for the first time in detail using light, scanning and transmission electron microscopy. The mature spermatozoa are of the bilaterian primitive type, also called aquasperm and develop as uniflagellate sperm consisting of a round head with distinct mitochondria at the base and a 9+2 flagellum of approximately 42 μm in length. The acrosomal complex consists of a small, round electron translucent acrosomal vesicle and a subacrosomal base. There is no separate midpiece, and the mitochondria surround the proximal and distal centriole in the posterior part of the head. The primitive structure of the spermatozoa suggests that these fertilize the egg by free spawning, probably the ancestral mode of fertilization in early bilaterians. When compared to the spermatozoa of other metazoans, we find that the arrangement of organelles in the Xenoturbella sperm shows similarities to a wide range of protostome and deuterostome taxa and does not seem to indicate any particular phylogenetic relationship. [ABSTRACT FROM AUTHOR]

The spermatozoa of two species of Macrodasyida (Gastrotricha), Urodasys anorektoxys and U. acanthostylis, show an ultrastructural organization diverging from one another and from other gastrotrichs: their main peculiarity is in the absence of mitochondria. In U. anorektoxys, the acrosome is a long, twisted column inserted into the nucleus, which is basally cylindrical, and the flagellum shows rows of peculiar, large globules parallel to the axonemal doublets. In U. acanthostylis, the acrosome is completely cork-screwed and surrounds the nucleus, and the tail shows columnar accessory fibers. At present, the absence of mitochondria in the mature sperm, and the peculiar fingerprint aspect of condensed chromatin are the only traits shared by the two species. The features of the spermatozoa of these two species of Urodasys widen the range of different models of gastrotrich spermatozoa, and place the genus in a peculiar position, from the spermatological point of view, within the Macrodasyida. The loss of mitochondria in mature spermatozoa is possibly related to either the dysoxic habitat of the two species or a peculiar fertilization mechanism. [ABSTRACT FROM AUTHOR]

Abstract: The fertility of three bulls carrying different Robertsonian translocations (rob(1;29), rob(14;17) and rob(26;29)) was evaluated. Oocytes-cumulus complexes obtained from slaughterhouse-derived ovaries were matured and then fertilised in vitro with frozen/thawed seminal material from the above mentioned subjects, and from control bulls with normal karyotype. An assessment was first made of the concentration, vitality and acrosome integrity of the seminal material to be sure that possible differences in the results of the in vitro fertilisation experiments were not due to seminal material quality. The results of the experiments, evaluated by the percentage of cleaved embryos and blastocysts per cleaved embryo, indicated that the three bulls carrying Robertsonian translocations had similar fertilising power and semen qualitative parameters to the controls. These data suggest that neither gametogenesys impairment nor decreased spermatozoa fertilising capacity is responsible for the reduced fertility in bulls with Robertsonian translocations. What the data do confirm is that the observed in vivo hypofertility for karyologically abnormal bulls is mainly due to early embryonic mortality. [Copyright &y& Elsevier]

Using two-dimensional gel electrophoresis, changes in protein phosphorylation caused by cyclic nucleotide-dependent protein kinases were analyzed with or without exposure to a protein kinase inhibitor, H-8, during the mouse sperm acrosomal reaction. The acrosomal reaction, induced by the treatment of sperm with dibutyryl cyclic AMP or dibutyryl cyclic GMP, was inhibited by H-8. The activities of cyclic AMP-dependent protein kinase (PKA) and cyclic GMP-dependent protein kinase induced by the sperm extract were also inhibited by H-8. When endogenous PKA in sperm was activated by the addition of cyclic AMP, a 45-kDa protein spot identified by electrophoresis indicated the occurrence of phosphorylation in vivo. Furthermore, the enhanced phosphorylation of the 45-kDa protein spot was inhibited by H-8. These results suggest that the PKA-catalyzed phosphorylation of the 45-kDa protein may be involved in the regulation of the mouse sperm acrosomal reaction. [ABSTRACT FROM AUTHOR]

Bisphenol A (BPA) is a global environmental contaminant that has been implicated as a potential endocrine disruptor. In the present study, newborn rats and mice were injected subcutaneously with BPA to determine the potential developmental effects on the testis. Testes were examined by light and electron microscopy at 15 weeks of age. Other groups of newborn mice and rats were injected with 17β-estradiol (E2) or β-estradiol 3-benzoate (E2B) in a similar manner. BPA, E2, and E2B had similar effects on testes. When treated animals reached puberty and spermiogenesis began, the first sign of the effects was detected in the steps 2–3 spermatids: the acrosomal granule and nucleus were deformed. Henceforth, abnormalities in the acrosome and nucleus were observed in older spermatids and spermatozoa. Ectoplasmic specialization between the Sertoli cell and spermatids was also affected: some specializations were partially or totally deleted. When animals fully matured, the effects of the agents were not found in the testes, and the animals were found to be fertile. The results of the present study show that BPA acts as an estrogen, and causes changes which appear to revert in adults. [Copyright &y& Elsevier]

Capacitated mammalian spermatozoa undergo a fusion response of their head plasma membrane and the outer acrosomal membrane leading to vesiculation classically known as acrosome reaction. Acrosome reaction occurs in response to various acrosome reaction inducers including zona pellucida proteins, calcium ionophore, dibutyryl cAMP, progesterone, etc. All the acrosome reaction inducers cause a transient of calcium influx into the sperm through voltage-dependent cation channels. Efflux of chloride, stimulation of activity of phospholipases, and phosphorylation of proteins are other known changes introduced by acrosome reaction inducers. Macromolecular organization and dynamics of sperm membranes during the progression of this vesiculation are largely unexplored. In this study, we report that progesterone induced the formation of horizontal microdomains within the exofacial surfaces of sperm membranes, which showed progressive and independent alterations in molecular dynamics. In the light of this observation, we propose that sperm membrane rafts may contain both horizontal and vertical microdomains. [Copyright &y& Elsevier]

Various signalling pathways are involved in the regulation of sperm motility, capacitation, acrosome reaction and sperm– zona binding. Recent data pointed out an important role for phosphatidylinositol 3-kinase (PI3K) in human sperm motility. However, no study as of yet has been carried out to determine the effect of sperm treatment with the PI3K inhibitor LY294002 on other sperm parameters. In the present study, we investigated the role of PI3K on human sperm motility, acrosome reaction and sperm–oocyte binding by using this inhibitor. We demonstrate that in vitro incubation of washed unselected spermatozoa with LY294002 increased the percentage motility and progressive motility in asthenozoospermia patients as evaluated by computer-aided sperm analysis. The compound furthermore did not influence the acrosome reaction, whilst it (further) slightly enhanced sperm-oocyte binding. Our results therefore imply that PI3K negatively affects sperm motility and oocyte binding and might suggest a possible therapeutic role for PI3K inhibitors in the treatment regime for asthenozoospermia. [ABSTRACT FROM AUTHOR]

BACKGROUND: Aggressive immobilization of sperm prior to ICSI significantly improves fertilization rates, but the mechanism of this effect is not yet clear. This study was performed in order to assess the characteristics of mechanically immobilized human sperm by transmission electron microscope (TEM). METHODS: Sperm obtained from ejaculated semen samples from three different donors were immobilized in a standard manner for ICSI. They were then injected into the perivitelline space of mouse oocytes in order to be able to locate them by TEM. Intact motile sperm injected subzonally served as controls (n = 160). Finally, the 'carrier' oocytes were fixed and processed for TEM. RESULTS: A total of 300 sperm were mechanically immobilized and inserted into the perivitelline space of mouse oocytes. Ultrathin sections revealed consistent alterations in the acrosomal region including disruption of the plasma membrane, and disruption, vesiculation or even loss of the acrosome. Thus, all of the sperm assessed had undergone some disorganization of the head, in contrast to a majority of control sperm. CONCLUSIONS: Immobilization of sperm for ICSI by compressing and rolling the sperm tails induces a variable disruption and sometimes loss of the acrosome. This could well be a reason for the higher success rates when ICSI is performed using immobilized sperm. [ABSTRACT FROM AUTHOR]

We previously demonstrated that Xenopus sperm undergo an acrosome reaction on the vitelline envelope (VE) in response to the materials secreted from the oviductal pars recta [Dev. Biol. 243 (2002), 55]. A monoclonal antibody against the acrosome reaction-inducing substance in Xenopus (ARISX) was obtained by immunizing mice with pars recta extract (PRE). The acrosome reaction by PRE or on the VE was effectively inhibited by the intact anti-ARISX antibody as well as its Fab fragment, indicating that the antibody recognizes the epitopes localized on the acrosome reaction-inducing substance. On Western blots, the anti-ARISX antibody recognized a molecule with an apparent molecular mass of 300 kDa in PRE and in the VE, but this molecule was not detected in the coelomic envelope. The amount of ARISX in PRE was increased by the treatment of females with pregnant mare serum gonadotropin. Periodate oxidation of PRE completely abolished the acrosome reaction-inducing activity, indicating the involvement of the carbohydrate moieties of ARISX in inducing the acrosome reaction. On immunofluorescence observation, ARISX was localized in the epithelial cells in the posterior region of the pars recta and on the VE as fibrous structures. [Copyright &y& Elsevier]

In the mouse and several other species, including man, capacitated acrosome-intact spermatozoa interact with natural [soluble zona pellucida (ZP) and progesterone (P4)] and synthetic [neoglycoproteins (ngps) and calcium (Ca2+) ionophore] agonists, prior to the initiation of a Ca2+-dependent signal transduction cascade. The net result is the fusion of the sperm plasma membrane overlying the outer acrosomal membrane at multiple sites and exocytosis of acrosomal contents [i.e., induction of the acrosome reaction (AR)]. This step is believed to be a prerequisite that enables the acrosome-reacted spermatozoon to penetrate the ZP and fertilize the egg. Although the rat is one of the most commonly used laboratory animals, very little is known about the chemical nature of agonists that induce the AR in this species. The lack of this information is primarily due to the fact that the rat sperm acrosome is a relatively thin structure. Thus, it is difficult to assess the status of the sperm acrosome in this species. In this report, we describe the use of a Coomassie brilliant blue dye staining procedure to assess the status of the rat sperm acrosome by light microscopy. The procedure is highly reproducible and has allowed us to determine the effects of carbohydrate (ngps and mouse ZP) and noncarbohydrate (P4 and Ca2+ ionophore) agonists on capacitated spermatozoa. In addition, we have used a pharmacological approach to examine the functional significance of calmodulin (CaM), a Ca2+-binding protein, in induction of the AR in spermatozoa. Data presented in this report demonstrate that several ngps, solubilized mZP, P4, and Ca2+ ionophores induce the AR in rat spermatozoa. Furthermore, we demonstrate that, whereas CaM antagonists blocked P4-induced AR, most of the inhibitors used had no significant effect on the Ca2+ ionophore-induced (nonphysiological) AR. [Copyright &y& Elsevier]

Binding to the egg''s zona pellucida stimulates the spermatozoon to undergo acrosome reaction, a process which enables the sperm to penetrate the egg. Prior to this binding, the spermatozoa undergo in the female reproductive tract a series of biochemical transformations, collectively called capacitation. The first event in capacitation is the elevation of intracellular calcium and bicarbonate to activate adenylyl cyclase (AC) to produce cyclic-AMP, which activates protein kinase A (PKA) to phosphorylate certain proteins. During capacitation, there is also an increase in actin polymerization and in the membrane-bound phospholipase C (PLC). Sperm binding to zona-pellucida causes further activation of cAMP/PKA and protein kinase C (PKC), respectively. PKC opens a calcium channel in the plasma membrane. PKA together with inositol-trisphosphate activate calcium channels in the outer acrosomal membrane, which leads to an increase in cytosolic calcium. The depletion of calcium in the acrosome will activate a store-operated calcium entry mechanism in the plasma membrane, leading to a higher increase in cytosolic calcium, resulting in membrane fusion and acrosome reaction. [Copyright &y& Elsevier]

Examines the zona pellucida for the induction of acrosome reaction in Germany. Evaluation of acrosomal status from sperm samples; Dependence of acrosome reaction induced by solubilized zona proteins; Effect of incubation on the spermatozoa in zonae.

Examines the effect of angiotensin converting enzyme (ACE) and angiotensins on human sperm functions. Control of the female and male reproductive functions by the renin angiontensisn system; Effect of ACE ont he capacitation process/acromosome reaction; Negative usefulness of the ACE release for clinical marker on human sperm capacitation.

The mature spermatozoon of Spadella cephaloptera belongs to the filiform type. At the front end it contains a membraneous cap, which may be an acrosomal structure, and some beaded sacs. A crystal structure is found in these sacs in spermatid stages. Near this front end there is a centriolar structure. A very elongated mitochondrial derivative runs through most of the sperm. In the central half of the sperm there is a structure composed of some longitudinal sacs lying alongside the axonema. The nucleus is situated in the hind end, just in front of a free flagellum. The spermatozoon is highly unique, but to some extent it agrees with spermatozoa found among arthropodes and gastropodes. [ABSTRACT FROM AUTHOR]

This study investigates the ability of sucrose to protect spermatozoa against mitochondrial damage, artificial cryoinduction of capacitation, and acrosome reaction. Spermatozoa were isolated using the swim-up procedure performed using three different media: (a) human tubal fluid (HTF, control) medium; (b) HTF with 1% human serum albumin (HSA); and (c) HTF with 1% HSA and 0.25 M sucrose. From each group, 30 µl suspensions of cells were dropped directly into liquid nitrogen and stored for at least 24 h. Cells were thawed by quickly submerging the spheres in HTF with 1% HSA at 37 °C with gentle agitation. Sperm motility, viability, mitochondrial membrane potential integrity, spontaneous capacitation, and acrosome reaction were investigated. Sperm viability, acrosome reaction, and capacitation were detected using the double fluorescence chlortetracycline-Hoechst 33258 staining technique. Mitochondrial function was evaluated using a unique fluorescent cationic dye, 5,5',6,6-tetrachloro-1-1',3,3'-tetraethyl-benzamidazolocarbocyanin iodide, commonly known as JC-1. The number of progressively motile spermatozoa was significantly higher in the sucrose-supplemented medium group (57.1±3.2%, P<0.05) when compared with controls (19.4±1.9%). The combination of HSA and sucrose (65.2±2.6%) has a stronger cryoprotective effect on the integrity of mitochondrial membrane potential (P<0.05) compared with HSA alone (32.6±4.7%). In conclusion, vitrification of human spermatozoa with non-permeable cryoprotectants such as HSA and sucrose can effectively cryopreserve the cells without significant loss of important physiological parameters. [ABSTRACT FROM AUTHOR]