• Asian-Australasian Journal of Animal Sciences
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• v.20 no.3
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• pp.351-358
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• 2007

Carbohydrate-protein interactions are known to be important in gamete interactions. Several evidence indicated that a fucose-containing sulfated polysaccharide fucoidan was potential inhibitor of fertilization in vitro and thus fucose seemed to be part of the recognition signal of gamete interaction in mammals. In recent investigation we found that ${\alpha}$-L-fucosidase activity was present in boar spermatozoa and it was related to sperm binding to and penetration into zona pellucida (ZP) in vitro. The objective of this study was to determine the effects of monosaccharide L-fucose and polysaccharide fucoidan on sperm ${\alpha}$-L-fucosidase activity and relation to sperm-oocyte interaction in pig. Results indicated that the activity of sperm ${\alpha}$-L-fucosidase was largely inhibited (62%) when sperm suspension was treated with monosaccharide L-fucose. It also significantly inhibited the number of sperm binding to ZP (32%) and penetration into zona-intact oocytes (72%), but did not inhibit penetration into zona-free oocytes when fertilization medium contained L-fucose. The chlorotetracycline (CTC) assessment showed that L-fucose did not affect induction of sperm capacitation and acrosome reaction. In contrast, the activity of sperm ${\alpha}$-L-fucosidase was not inhibited when sperm suspension was treated with polysaccharide fucoidan but sperm-ZP binding was greatly inhibited (85%) and completely blocked sperm penetration into zona-intact or zona-free oocytes. The CTC assessment showed that fucoidan increased the F pattern and decreased the AR pattern sperm. These results suggested that the different inhibitory mechanisms were present between monosaccharide L-fucose and polysaccharide fucoidan on sperm-oocyte interaction, the inhibition effect of ${\alpha}$-L-fucose on sperm binding and penetrating into ZP caused sperm ${\alpha}$-L-fucosidase inhibited by ${\alpha}$-L-fucose.

• Journal of Embryo Transfer
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• v.10 no.3
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• pp.243-250
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• 1995

Frozen storage of the oocytes has been used in a few mammalian species including mouse, hamster, human and cattle. However, frozen4hawed oocvtes show different sperm penetration on the levels of the zona pellucida and the plasma memhrane when compared with fresh oocytes. To elucidate biological changes occurring during freezing and thawing, we examined the kinetics of sperm penetration into frozen-thawed hamster oocytes. Oocytes obtained from superovulated female golden hamsters were frozen-thawed in an autofreezer according to an established method. Fresh and frozen4hawed oocytes were fertilized in vitro with capacitated hamster spermatozoa after removing the zona pellucida. The oocytes were examined at 1, 2, 3 and 6 h postinsemination. Sperm penetration found to be 1 h delayed in frozen-thawed oocytes. Other parameters such as degree of polyspermy and decondensing sperm heads were not affected by freezing and thawing. The results suggest that freezing and thawing may cause changes in the egg membrane surface and subsequently which leads to delay in the sperm-egg fusion.

• The Korean Journal of Zoology
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• v.28 no.1
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• pp.1-8
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• 1985

A series of observation on sperm penetration in urodele (Ambystoma mexicanum) eggs are reported. The whole sperm including the tail appears to penetrate the egg surface. It can be demonstrated that the Ambystoma mexicanum egg is typically polyspermic. Each sperm penetration point is marked by a distinct crater on the egg surface the so called sperm pit. Initially, no sign of disruption in the surface structure observed. Once sperm penetration was complete, the site of entry became covered with long microvilli.

• Clinical and Experimental Reproductive Medicine
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• v.16 no.2
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• pp.173-177
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• 1989

Defective or inadequate semen quality, usually presenting as low sperm count or poor sperm motility , is recognizable by semen analysis. However, the ability of spermatozoa to fertilize an ovum is not determined used in various experiments. In this study, hamster oocyte sperm penetration assay was used to determine the fertilizing capacity of sperms in 20 subjects which divided into two groups, group A with 10 normal fertile men, and group B with 10 infertile men. The % penetration in group A and group B were 61% and 35% respectively, which showed statistically not significant but fertilization index was significantly different between group A(FI=2.24) and group B(FI=O.05). Additionally it seemed that the percentage of sperm penetraton was influenced more by the motility of spermatozoa than by the number.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.1
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• pp.31-41
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• 1994

Morphological estimation of human spermatozoa is complicated by the fact that there is great natural variation in shape. This natural variation in shapes makes it difficult to say which forms are associated with infertility and which are normal variations. Possibly post coital test or in vitro cervical mucus penetration tests will help to clarify this question by showing which sperm are capable of penetration. The purpose of this investigation was performed to assess distribution of various morphological abnormalities according to the ability of sperm to penetrate cervical mucus. The sperm-mucus penetration using hen's egg white as substituting mucus for human cervical mucus was done in 45 fertile men with normal semen analysis and 122 infertile men with abnormal seminal parameters more than one. The female partners of 122 infertile couples showed normal results in the female fundamental test for fertility. Conventional semen analysis was evaluated according to the WHO standard normal(l980). The detailed classification of the abnormal sperm was made according to David et al(l975). The vitality of the sperm samples determined by eosin yellow-nigrosin stainig according to the method of Eliasson(l977). Results were as follw; 1. The patients had significantly lower total sperm count, motility (%), normal morphology (%), viability and total functional sperm fractions(TFSF) than fertile donors. 2. The mean value of sperm penetration distance of the patients(28.69${\pm}$11.02mm) showed significantly lower than fertile donors(37.33${\pm}$5.49mm). And 43/45 fertile donors(95.5%) as well as 57/122 patients(46.7%) had over 30mm in sperm penetration distance respectively. While 2/45 fertile donors(4.5 %) and 65/122 patient(53.3%) had under 30mm in sperm penetration distance respectively. 3. The morphological abnormalities in fertile donors were significantly lower 23.04${\pm}$5.83% (head = 12.89${\pm}$4.98, neck=6.11${\pm}$3.83%, and tail=3.43${\pm}$2.65%), compared to 36.03${\pm}$14. 40% in patients(head = 15.98 8.60%, neck 11.20${\pm}$6.56% and tail=8.70${\pm}$6.55%). Also, 3 types of sperm abnormalities including head, neck and tail were significantly lower in patient than fertile donors, respectively. Both the patients and fertile donors showed higher distribution of sperm with abnormal head than abnormal neck and tail. 4. The mean morphological abnormalities(SP>30mm) of the patients(30.68 11.64%; head = 15.95${\pm}$9.35%, neck=8.14${\pm}$4.21 %, tail=6.56${\pm}$5.64%) were significantly lower compared to patients(40.72${\pm}$15.01 %; head=16.02${\pm}$7.69%, neck 13.89${\pm}$7.82%, tail=1O.58${\pm}$6.75%) under 30mm in sperm penetration distance. Also, both groups over 30mm and under 30mm in sperm penetration showed distance higher distribution of sperm with abnormal head than abnormal neck and tail. The morphological abnormalities of head did not show significant difference but abnormal neck and tail were significant difference between the over 30mm and under 30mm group in sperm penetration distance.

• Environmental Analysis Health and Toxicology
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• v.19 no.2
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• pp.177-182
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• 2004

Alcohol consumption and exposure to endocrine disruptors and industrial solvents have been implicated in impaired spermatogenesis, increase in the incidence of malformed sperm and decrease in the percentage of moving sperm. The aim of this study was to determine and compare the direct effects of some organic solvents(bisphenol A; BPA, dibutyl phthalate; DBP, formaldehyde; HCHO, dimethylsulphoxide; DMSO, ethanol) on mucus penetration distance, motility and survival rate of human sperm in vitro. Semen samples from 3 health subjects were prepared using swim-up method and 0.0005-0.5% organic solvents were added to the test medium. BPA, DBP, HCHO and DMSO produced significant decreases in the motility and survival rate with a different potency. The most potent inhibition of mucus penetration distance, motility and survival rate was observed after exposure to HCHO. A concentration of 0.0005% HCHO significantly inhibited sperm motility. When ethanol m.: added directly to sperm, at concentrations equivalent to that in serum after heavy drinking, these damaging effects were lowest compared with other solvents. Present study shows that each compound has different toxic potency to human sperm and we need special caution for the use of HCHO.

• Korean Journal of Veterinary Research
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• v.32 no.3
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• pp.435-450
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• 1992

To evaluate the fertilizing capacity of domestic animal spermatozoa by hamster test, semen were collected from 15 boars(Duroc, Landrace, and Yorkshire) and 2 mixed dogs which had been proved to be fertile in the past then, the semen were preserved in BWW medium at $4^{\circ}C$ or $18^{\circ}C$ for about 20 hours and coincubated with zona-free hamster ova for 5 hours. The ova were stained by lacmoid and examined under phase contrast microscope to investigate the rates of sperm binding to the ova, penetration and formation of a male pronucleus, and the numbers of both bound and penetrated sperm per ovum. Both the semen preserved at $18^{\circ}C$ for about 20 hours and that treated by swim up procedure showed considerably higher rates of sperm binding and penetration as well as higher number of penetrated sperm than that preserved at $4^{\circ}C$ for about 20 hours, respectively(p<0.01). Motility of boar sperm at insemination was from 40 to 90% and no difference in hamster test was obtained according to different degree of sperm motility. Abnormality in morphology of boar sperm at insemination was from 6 to 45% and no difference in hamster test was obtained according to different degree of sperm abnormality. The sperm concentrations of $7{\times}10^7$ and $7{\times}10^6$ showed considerably higher rates of sperm binding and penetration as well as higher number of bound sperm than that of $7{\times}10^4$ (p<0.01) along with the same higher results than that of $7{\times}10^5$(0<0.05), respectively. Boar sperm showed considerably higher rates of sperm binding and penetration as well as higher numbers of both bound and penetrated sperm than dog sperm, when both semen were treated by BWW＋heparin medium and swim up procedure, respectively. These results indicated that fertile boar sperm showed considerably lower rates in the results of hamster test, when preserved at $4^{\circ}C$ for about 20 hours and in lower concentration of sperm than when preserved at $18^{\circ}C$ for about 20 hours and in higher concentration of sperm, respectively, and at the same time considerably higher results than fertile dog sperm, consequently to prove that hamster test would be of great value in assaying the fertilizing capacity of boar sperm.

• Journal of Veterinary Clinics
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• v.10 no.1
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• pp.1-10
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• 1993

To investigate the availability of hamster test in assaying the fertilizing capacity of dog sperm and the effect of canine sperm motility on sperm binding and penetration, semen were collected from four dogs(three dogs had been proven to be fertile and one dog to be subfertile during the past two years) and then preserved in BWW(Biggers, Whitten, Whittingham) medium for about 20 hours. The semen were given each different treatment according to the experimental design and coincubated with zona-free hamster ova for 5 hours. The ova were stained by lacmoid and examined under phase contrast microscope to investigate the rates of ova bound with sperm(sperm binding) and ova penetrated by sperm(penetration), and also numbers of both bound and penetrated sperm per ovum. In comparison between fertile dogs and a subfertile dog, the rate of sperm binding was higher in fertile dogs than the subfertile dog(p<0.01, p<0.05). The number of bound sperm per ovum was considerably higher in a fertile dog than the subfertile dog((p<0.01), and also difference of number of the bound sperm was obtained among the fertile dogs(p<0.01, p<0.05). The rate of penetration as well as the number of penetrated sperm per ovum was higher in the fertile dogs than the subfertile dog(p<0.01, p<0.05). In fertile dogs. the canine semen preserved at 4$^{\circ}C$ for 18 to 22 hours showed from 30 to 80% motility at Insemination, however, no difference in hamster test was obtained according to different degree of sperm motility. These results indicated that hamster test would be of avail in assaying the fertilizing capacity of dog sperm.

• Clinical and Experimental Reproductive Medicine
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• v.19 no.1
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• pp.57-64
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• 1992

The present study was undertaken to clarify the role of TEST-Yolk Buffer(TYB) as a factor for the improvement of human sperm fertility potential. We examined the effects of low temperature capacitation using TYB on sperm motility (%), motility pattern, normal morphology, true acrosome reaction, sperm penetration assay and human in vitro fertilization. Comparing the TYB method and swim-up method, the sperm motility(%) of selected sperm was not significantly different, but statistically significant differences were found in curvilinear velocity, linearity, lateral head displacement, normal morphology(%) and true acrosome reaction(%)(p<0.05). Results obtained from the sperm penetration assay demonstrated that the penetration index and penetration rate were increased significantly(p<0.05) when the spermatozoa were incubated in TYB, as compared with swim-up method. And fertilization of intact human oocytes was more succesful when spermatozoa were pretreated with TYB at $4^{\circ}C$ for 48 hours as compared with swim-up method. Our results show that TYB method have advantages in terms of enhancement of sperm hyperactivation, increased true acrosome reaction, increased ability to penetrate zona-free hamster ova and augmented fertilization of human oocytes, suggesting that TYB is superior in its ability to preserve sperm motility and fertilizing ability.

• Journal of Veterinary Clinics
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• v.9 no.2
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• pp.373-390
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• 1992

To assay the fertilizing capacity of domestic animal spermatozoa by hamster test, semen were collected from 13 boars(Duroc. Landrace and Yorkshire) which had been proved to be fertile in the past. then, were preserved in BWW medium or in raw state at 18$^{\circ}C$ or at room temperature. The preserved semen were given each different treatment according to the experimental design and coincubated with zona-free hamster ova for 5 hours. The ova were stained by lacmoid and examined under phase contrast microscope to investigate the rates of ova bound with sperm(sperm binding). ova penetrated by sperm(penetration) and formation of a male pronucleus(pronucleus formation) and also numbers of both bound and penetrated sperm per ovum. Between BWW and TBM medium for boar sperm. no difference in the results of hamster test was obtained. The boar spermatozoa in BWW medium, BWW with caffeine, BWW with heparin, and BWW with both caffeine and heparin showed no difference in the results of hamster test. The boar spermatozoa in BWW medium containing both calcium and RSA showed considerably higher rates of sperm binding, penetration and pronucleus formation as well as higher numbers of both bound and penetrated sperm than those not containing calcium with or without BSA( p<0.01) and also the same results higher than that containing calcium without BSA( p< 0.05). The boar spermatozoa irradiated by X-ray(70 KVP, 20mA) for 3 seconds. then, maintained at 18$^{\circ}C$ for 18 hours showed considerably lower rate of sperm binding than all the other groups including the control and X-ray groups irradiated by smaller dose or maintained for shorter period(p<0.01), and also showed lower number of bound sperm than the other groups(p<0.01, p<0.05). All the control groups of both raw and diluted sperm in BWM medium showed higher rates of sperm binding, penetration and pronucleus formation as well as higher number of penetrated sperm than all the X-ray groups irradiated for 3 seconds(70KVP, 20mA) and maintained for either 3 or 18 hours (p<0.01, p<0.05). At the same time the control groups of diluted sperm showed considerably higher rates of sperm penetration and pronucleus formation than the control group of raw sperm( p<0.01). These results indicates that fertile boar sperm showed considerably lower rates In the results of hamster test, when incubated in the medium without calcium and irradiated by X-ray than when incubated in the medium with calcium and not irradiated by X-ray, respectively, to prove consequently that hamster test would be of great value in assaying the fertilizing capacity of boar spermatozoa.