Evaluation of acrosome reaction and viability of human sperm with two fluorescent dyes

Ozaki, Tomoya; Takahashi, Kentaro; Kanasaki, Haruhiko; Miyazaki, Kazuhiko

doi:10.1007/s004040000112

Cited by 20 publications

(16 citation statements)

References 7 publications

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“…There are some studies describing the lectin‐binding proteins involved in spermatozoa capacitation and AR (Fierro et al. , 1996; Ozaki et al. , 2002).…”

Section: Discussionmentioning

confidence: 99%

Con A‐binding protein Zn‐α2‐glycoprotein on human sperm membrane is related to acrosome reaction and sperm fertility

et al. 2011

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Fertilization, the recognition and fusion between spermatozoa and oocyte, involves various molecules on the spermatozoa and oocyte membranes. Concanavalin A (ConA)-binding proteins may be one of the molecules involved in mammal spermatozoa fertilization; however, their structure and function remain largely unknown. Here, we initially identified a ConA-binding protein, Zn-α2-glycoprotein (ZAG), involved in regulating the acrosome reaction (AR) of human spermatozoa. ZAG is localized on the pre-equatorial region covering the acrosome, neck and tail (some parts of middle piece and principal piece respectively) regions of the acrosome intact human spermatozoa, and disappears in the acrosomal region of the acrosome-reacted spermatozoa. Polyclonal antibodies against human recombinant ZAG significantly reduced the AR and sperm capability binding to human zona pellucida or penetration into zona-free hamster oocytes. Furthermore, assessment of the signaling pathways regulated by ZAG revealed that ZAG affects sperm AR through both the cAMP/PKA and PKC pathways. These results indicate that ZAG, which is present on the human sperm membrane, plays a critical role in the AR and subsequently, may be involved in sperm fertility.

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“…There are some studies describing the lectin‐binding proteins involved in spermatozoa capacitation and AR (Fierro et al. , 1996; Ozaki et al. , 2002).…”

Section: Discussionmentioning

confidence: 99%

Con A‐binding protein Zn‐α2‐glycoprotein on human sperm membrane is related to acrosome reaction and sperm fertility

et al. 2011

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“…The acrosomal integrity was assessed according to the procedures described by Santos et al [18] and Ozaki et al [19] using a fluorescence inverted microscope (OLYMPUS BX53, Japan). One drop of sperm suspension was placed on a microscopic slide, and through the fluorescence labeling of the sperm nuclei with Hoechst 33258 and acrosomes with FITC–PSA, we examined the integrity of the sperm on days 15, 60 and 90.…”

Section: Methodsmentioning

confidence: 99%

Exposure to Silica Nanoparticles Causes Reversible Damage of the Spermatogenic Process in Mice

Wang

et al. 2014

PLoS ONE

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Environmental exposure to nanomaterials is inevitable, as nanomaterials have become part of our daily life now. In this study, we firstly investigated the effects of silica nanoparticles on the spermatogenic process according to their time course in male mice. 48 male mice were randomly divided into control group and silica nanoparticle group with 24 mice per group, with three evaluation time points (15, 35 and 60 days after the first dose) per group. Mice were exposed to the vehicle control and silica nanoparticles at a dosage of 20 mg/kg every 3 days, five times over a 13-day period, and were sacrificed at 15, 35 and 60 days after the first dose. The results showed that silica nanoparticles caused damage to the mitochondrial cristae and decreased the levels of ATP, resulting in oxidative stress in the testis by days 15 and 35; however, the damage was repaired by day 60. DNA damage and the decreases in the quantity and quality of epididymal sperm were found by days 15 and 35; but these changes were recovered by day 60. In contrast, the acrosome integrity and fertility in epididymal sperm, the numbers of spermatogonia and sperm in the testes, and the levels of three major sex hormones were not significantly affected throughout the 60-day period. The results suggest that nanoparticles can cause reversible damage to the sperms in the epididymis without affecting fertility, they are more sensitive than both spermatogonia and spermatocytes to silica nanoparticle toxicity. Considering the spermatogenesis time course, silica nanoparticles primarily influence the maturation process of sperm in the epididymis by causing oxidative stress and damage to the mitochondrial structure, resulting in energy metabolism dysfunction.

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“…At least 200 sperm from each mouse were examined. Acrosomal status was assessed according to the procedures of Ramalho-Santos45 and Ozaki46.…”

Section: Methodsmentioning

confidence: 99%

Repeated administrations of carbon nanotubes in male mice cause reversible testis damage without affecting fertility

et al. 2010

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Soluble carbon nanotubes are promising materials for in vivo delivery and imaging applications. Several reports have described the in vivo toxicity of carbon nanotubes, however, their effects on male reproduction have not been examined. Here we show that repeated intravenous injections of water-soluble multi-walled carbon nanotubes into male mice can cause reversible testis damage without affecting fertility. Nanotubes accumulated in the testes, generated oxidative stress, and decreased the thickness of the seminiferous epithelium in the testis at day 15, but the damage was repaired after 60 and 90 days. The quantity, quality, and integrity of the sperm and the levels of three major sex hormones were not significantly affected throughout the 90-day period. The fertility of treated male mice was unaffected; the pregnancy rate and delivery success of female mice that mated with the treated male mice did not differ from those that mated with untreated male mice.

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Evaluation of acrosome reaction and viability of human sperm with two fluorescent dyes

Abstract: Viability and acrosome status of a human sperm can be easily assessed simultaneously by Hoechst 33258/FITC-PSA staining method. This combination method is considered useful to evaluate sperm function.

Cited by 20 publications

References 7 publications

Con A‐binding protein Zn‐α2‐glycoprotein on human sperm membrane is related to acrosome reaction and sperm fertility

Con A‐binding protein Zn‐α2‐glycoprotein on human sperm membrane is related to acrosome reaction and sperm fertility

Exposure to Silica Nanoparticles Causes Reversible Damage of the Spermatogenic Process in Mice

Repeated administrations of carbon nanotubes in male mice cause reversible testis damage without affecting fertility

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