SNAREs in Mammalian Sperm: Possible Implications for Fertilization

Ramalho‐Santos, João; Moreno, Ricardo D.; Šutovský, Peter; Chan, Anthony W.S.; Hewitson, Laura; Wessel, Gary M.; Simerly, Calvin; Schatten, Gerald

doi:10.1006/dbio.2000.9745

Cited by 107 publications

(81 citation statements)

References 58 publications

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“…The fact that blocking antibodies against sperm surface antigens did not completely impair fertilization was also observed in our previous assays with bovine sperm (Cesari et al 2004b) and by other authors (Ramalho-Santos et al 2000). Although the physiological interpretation of IVF results in the presence of antibodies against specific proteins is controversial, the role of sperm and oocyte surface proteins has been determined by this experimental approach (Hasegawa et al 2000, Peknicova et al 2001.…”

Section: Discussionsupporting

confidence: 78%

Serine protease activity, bovine sperm protease, 66 kDa (BSp66), is present in hamster sperm and is involved in sperm–zona interaction

Cesari

Katunar²,

Monclus³

et al. 2005

Reproduction

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Bovine sperm protease, 66 kDa (BSp66) is a serine protease previously characterized in bovine spermatozoa. Like other proteases, it may be present in sperm from other mammalian species different from bovine, playing a role in the fertilization process. In this study, we looked for BSp66 in hamster spermatozoa using heterologous antibodies against bovine BSp66. An immunoreactive protein was detected by Western blotting in mature and immature sperm. The detected protein had two isoforms similar to the ones reported in bovine sperm. Furthermore, indirect immune detection by fluorescence and electron microscopy assays, showed BSp66 signal at the acrosomal region similar to bovine sperm. As it was determined in bovine sperm, the acrosomal reaction displays the antigen within the acrosomal content. When live hamster sperm was incubated with polyclonal antibody against bovine BSp66 a decrease in the number of sperm bound to zona pellucida in homologous IVF and an impairment of head–head agglutination, were observed. These results suggest that a protease homologous to bovine BSp66 is present in golden hamster spermatozoa, with a conserved molecular mass and cellular location. Moreover, hamster BSp66 is probably involved in zona pellucida recognition.

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Section: Discussionsupporting

confidence: 78%

Serine protease activity, bovine sperm protease, 66 kDa (BSp66), is present in hamster sperm and is involved in sperm–zona interaction

Cesari

Katunar²,

Monclus³

et al. 2005

Reproduction

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“…Frozen samples were thawed in warm modified Tyrodelactate medium with pyruvate and albumin, and immunocytochemistry using VAMP antibodies (a kind gift of Dr. Gary Wessel, Brown University; and obtained commercially via Synaptic Systems, Göttingen, Germany) was carried out as described in detail elsewhere (1). Briefly, poly-L-lysine coated coverslips with attached sperm were fixed in 2% formaldehyde, permeabilized in 1% Triton X-100, and blocked by further incubation in phosphate-buffered saline (PBS) containing 2 mg/ml bovine serum albumin and 100 mM glycine.…”

Section: Methodsmentioning

confidence: 99%

“…One protein that has been recently identified on the human acrosome is the vesicle-associated membrane protein (VAMP), also known as synaptobrevin (1). Together with other acrosomal components, VAMP may have an important functional role in promoting membrane fusion during the acrosome reaction (1).…”

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confidence: 99%

VAMP/synaptobrevin as an acrosomal marker for human sperm

Ramalho‐Santos

Schatten

2002

Fertility and Sterility

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Objective:To determine the possible use of the mammalian acrosomal marker vehicle-associated membrane protein (VAMP)/synaptobrevin to detect acrosome abnormalities in human sperm. Design: Analysis of human sperm after fixation and staining with an anti-VAMP antibody. Setting: An academic research institution. Patient(s): Semen samples from consenting patients who were participating in an infertility treatment program. Intervention(s): Human sperm samples were fixed, permeabilized with detergent, and examined by immunocytochemistry. Main outcome measure: Immunostaining. Result(s):Observation of sperm from patients with no obvious sperm morphological defects revealed normal looking acrosomes, as assessed by VAMP immunostaining. However, severe acrosome malformations were detected in other cases. The observations registered varied from the absence of a fully formed organelle in samples of patients with globozoospermia to abnormal VAMP staining in samples from patients with acrosomal defects. Conclusion: VAMP/synaptobrevin may be a useful marker for the functional assessment of acrosomal status in human sperm. (Fertil Steril 2002;77:159 -61.

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“…The increase in intracellular calcium is necessary for docking and fusion of the outer acrosomal and plasma membranes during acrosomal exocytosis. Membrane fusion appears to include SNAREs, Rab3A, and other molecules involved with membrane fusion in somatic cells (Schulz et al, 1997;Iida et al, 1999;Ramalho-Santos et al, 2000).…”

Section: Signal Transduction Steps During the Acrosome Reactionmentioning

confidence: 99%

Molecular Basis of Mammalian Gamete Binding

Miller

Shi

Burkin

2002

Recent Progress in Hormone Research

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Despite the importance of fertilization for controlling human reproduction, regulating animal production, and promoting preservation of endangered species, the molecular basis underlying gamete binding and fertilization has been perplexing. More progress has been made in the mouse than in other mammals and, recently, targeted deletion of specific genes in the mouse has yielded intriguing results. This review will emphasize research performed by our laboratory and others done primarily with mouse gametes but will include some interesting observations from other mammals. Studies of murine fertilization indicate that oligosaccharides on the egg coat glycoprotein ZP3 bind sperm. The precise oligosaccharides that bind sperm are the subject of considerable debate. ZP3 also induces exocytosis of the sperm acrosome, allowing sperm to penetrate through the egg coat (zona pellucida). A number of candidate ZP3 receptors have been proposed and studies of ␤1,4galacto-syltransferase-I (GalT-I) are reviewed here in the most detail. Sperm from mice with a targeted deletion of GalT-I still are able to bind the zona pellucida but are unable to acrosome react and penetrate through the zona. Therefore, the unique role of GalT-I appears to be in signal transduction. GalT-I forms a complex with heterotrimeric G proteins and activates signaling, leading to exocytosis in sperm and in heterologous cells expressing GalT-I. Other signaling steps triggered by GalT-I are under active investigation; this receptor forms a complex with a protein kinase anchoring protein.After exocytosis of the acrosome, sperm penetrate the zona pellucida and fuse with the oocyte plasma membrane using ADAM family members on sperm and integrins on oocytes. These proteins, along with the tetraspanins on oocytes, may form a complex web at gamete fusion. Targeted deletion of specific genes in this putative complex has provided important information about their redundancy. After the oocyte is fertilized, the binding site for GalT-I is lost from ZP3, preventing additional sperm from binding to the zona pellucida. New technical advances and creative ideas offer the opportunity to make important advances and to solve the conundrum of fertilization.

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SNAREs in Mammalian Sperm: Possible Implications for Fertilization

Cited by 107 publications

References 58 publications

Serine protease activity, bovine sperm protease, 66 kDa (BSp66), is present in hamster sperm and is involved in sperm–zona interaction

Serine protease activity, bovine sperm protease, 66 kDa (BSp66), is present in hamster sperm and is involved in sperm–zona interaction

VAMP/synaptobrevin as an acrosomal marker for human sperm

Molecular Basis of Mammalian Gamete Binding

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