• Journal of Embryo Transfer
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• v.16 no.1
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• pp.61-68
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• 2001

Sperm competent for fertilization can become capacitated, bind to the zona pellucida (ZP) of an egg in a specific manner, and complete acrosomal exocytosis. Failure to carry out these functions results in infertility. Although the interactions between the ZP and the plasma mem-brane overlying the sperm acrosome have been considered important for sperm-egg recognition and signalling, recent results have prompted a reassessment of current paradigms concerning these interactions. In this review, we're going to discuss about the roles of the acrosomal matrix, the particulate component of the acrosomal contents, in fertilization. The general hypothesis is that acrosomal exocytosis leads to the exposure of acrosomal matrix proteins that become de-facto extracellular matrix (ECM) on the surface of the sperm head, and that the dynamic interactions of this newly-exposed sperm ECM with the egg ECM (the ZP) govern sperm-egg recognition and sperm penetration of the ZP Informations from these experiments may provide new ways to address the poor ZP binding of sperm from some human infertility patients and may offer new avenues for contraception through the disruption of purposeful sperm-ZP binding.

• Journal of Veterinary Clinics
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• v.21 no.4
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• pp.363-368
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• 2004

To investigate the effects of seeding during freezing procedure on post-thaw viability, motility, and acrosome integrity of boar spermatozoa, semen from 5 Yorkshire boars were collected for this experiment. Raw semen were diluted with Merck I, subsequently added with cooling diluent containing lactose and egg yolk and with freezing diluent containing glycerol. The diluted semen were frozen on the rack in the styrofoam box filled with liquid nitrogen at the distance of 5 cm or I cm above LN2 level. Seeding was performed to only a group of straws frozen at 5 cm away on the surface of LN2. The frozen semen were thawed in $50^{\circ}C$C water and the viability and local motility were analyzed by sperm analysis imaging system. A part of thawed semen was taken for the examination of morphology of apical ridge of the acrosome to compare with the effect of seeding between the seeding-treated and non treated groups. I. Post-thaw viability was considerably higher in seeding-treated sperm than non-seeding group (p<0.01), however, no difference of local motility was obtained among the groups. 2. At three hours after thawing, viability was also higher in seeding-treated group than non-treated group (p<0.05), along with no difference of motility among the groups. 3. Higher normal acrosome integrity was obtained in the seeding-treated sperm than non-treated groups (p<0.01). 4. Between non-seeded groups, higher normal acrosome integrity was obtained in the sperm group frozen at 5cm upper on the surface of LN2 than that frozen at 1cm away (p<0.01). These results indicated that seeding treatment during freezing boar spermatozoa was beneficial to post-thaw viability and normal acrosome integrity.

• Korean Journal of Animal Reproduction
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• v.26 no.4
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• pp.339-345
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• 2002

The present study was carried out to investigate the effects of season on semen characteristics, frozen-thawed sperm viability and testosterone concentration in Yorkshire boars. There were no significant differences in the semen volume and sperm concentration on Yorkshire boars among spring, summer, autumn and winter. However, the pH of sperm-rich and sperm-poor fractions in winter season was higher than in spring, summer and autumn season in Yorkshlre boars. Sperm motiliy and normal acrosome of raw semen in Yorkshire boars did not differ significantly among spring, summer, autumn and winter. However, motility and normal acrosome of frozen-thawed sperm were higher in spring season than in summer, autumn and winter. Serum testosterone concentrations in Yorkshire were higher in spring than summer, autumn and winter. In conclusion, we found out that serum testosterone concentrations were very important role for frozen-thawed sperm viability in Yorkshire boars.

• Animal cells and systems
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• v.1 no.2
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• pp.371-379
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• 1997

To investigate the movement of sperm head and the role of sperm neck in forward sperm motility in the Korean striped field mouse, Apodemus agrarius coreae, the morphological characteristics of the cauda epididymal spermatozoa were examined by light microscopy and scanning and transmission electron microscopy. Spermatozoa of A. agrarius coreae were characterized by the conspicuous shape of the acrosome and the long tail compared with those of other rodents. Total length of the sperm was $133\mu{m}$. The sperm head had a curved falciform shape. The head was 8.0${\mu}$m in length, and about 4.0 ${\mu}$m in width. The shape of acrosome had an openerlike form. The sperm tail (125 ${\mu}$m) consisted of four major segments: neck (0.5 ${\mu}$m), middle piece (29.5 ${\mu}$m), and principal piece plus the end piece (95 ${\mu}$m). The outer dense fibers were arranged in a horseshoe fashion, and No. 1, 5, 6, and 9 of the outer dense fibers were larger than the others. The mitochondrial bundles of middle piece were composed of a pair of arms, which surrounded the axone of the middle piece by the 45 0 angled helical structure. The total number of mitochondrial gyres was 188. In particular, the microfilament structures existed in plasma membrane of the sperm, which was adjacent to the acrosomal region on the nuclear membrane. The segmented columns were surrounded by microfilament structures, and the microfilament bundles were adjacent to the outer membrane of the first mitochondria of middle piece. This study presents for the first time the existence of microfilament structures within the plasma membrane of sperm which is located from the adjacent acrosome region to the connecting piece in sperm neck of Korean striped field mouse, Apodemus agrarius coreae. The present result suggests that the constriction and extension of microfilament in sperm neck as well as the wave-movement of sperm tail may play a role in the movement of sperm head.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.10
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• pp.1369-1373
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• 2004

The percentages of sperm motility and normal acrosome on the liquid boar semen diluted and preserved at $4^{\circ}C$ with lactose hydrate, egg yolk and N-acetyl-D-glucosamine (LEN) diluent were significant differences according to preservation day and incubation time, respectively. The sperm motility steadily declined from 96.9% at 0.5 h incubation to 78.8% at 6 h incubation at 1 day of preservation. However, the sperm motility rapidly declined after 4 day of preservation during incubation. The normal acrosome steadily declined from 93.3% at 0.5 h incubation to 73.8% at 6 h incubation at 1 day of preservation. However, the normal acrosome rapidly declined after 3 day of preservation during incubation. The rates of sperm penetration and polyspermy were higher in 5 and $10{\times}10^6$ sperm/ml than in 0.2 and $1{\times}10^6$ sperm/ml. Mean numbers of sperm in penetrated oocyte were highest in $10{\times}10^6$ sperm/ml compared with other sperm concentrations. The rates of blastocysts from the cleaved oocytes (2-4 cell stage) were highest in $1{\times}10^6$sperm/ml compared with other sperm concentrations. In conclusion, we found out that liquid boar sperm stored at $4^{\circ}C$ could be used for in vitro fertilization of pig oocytes matured in vitro. Also, we recommend $1{\times}10^6$sperm/ml concentration for in vitro fertilization of pig oocytes.

• The Korean Journal of Zoology
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• v.38 no.4
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• pp.514-522
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• 1995

In order to examine the interaction of albumin with the sperm during capacitation in mouse, proteins of cauda epididymal sperm were extracted under various conditions and analyzed with SDS-PAGE. Sperm surface labeling patterms were also examined using fluorochroin~conjugated wheat germ agglutinin (WGA) and bovine serum albumin (BSA). Albumin was detached from the sperm surface during the incubation and seemed to be constituted the major protein components of the conditioned media in which sperm incubated for 90 mm. Detachment of albumin from the sperm was not affected by the Ca2+ in the medium. WGA-FITC labeling confirmed that Triton X-100 permeabilired plasma membrane overlaying the apical segment of sperm head and detached plasma membrane associated proteins having negatively charged glycoconjugates. BSA-FITC labeling of epididymal sperm occurred on the apical segment of periacrosoinal region and postacrosomal region of the head. BSA-FITC labeling was not observed in periacrosoinal region of the sperm treated with Ca2+-ionophore ~3187 (10 MM)~ whereas the postacrosome region of acrosome-reacted sperm was still labeled after the AR. These results suggest that albumin bound to the surface of epididymal sperm is detached during the capacitation process, and it might be involved In physiological change of sperm plasma membrane accompanying the capacitation.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.4
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• pp.486-494
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• 2006

In order to protect the spermatozoa against cold shock, hen egg yolk is widely used as a cryoprotective agent in semen freezing extenders for domestic animals. The protective action of yolk is largely presumed to be due to low density lipoproteins (LDL). The effects of LDL on sperm quality of bull and northern pike (Esox lucius) after freezing-thawing have been reported, but no study has been made to evaluate the effect of LDL on boar sperm motility and other characteristics. The experiment was carried out to investigate the effect of LDL on the freezing of boar sperm in 0.25 ml straws. The aim was to evaluate the quality of boar spermatozoa cryopreserved in the presence of LDL. Motility of semen cryopreserved in LDL was analyzed and compared to semen cryopreserved with Tris-citric acid-glucose (TCG) and Tris-citric acid-fructose (TCF), two basic freezing extenders containing egg yolk. Similarly, acrosome and plasma membrane integrity were also evaluated and compared to semen cryopreserved with TCG and TCF. Analysis of sperm quality after freeze-thaw showed that the motility, acrosome and plasma membrane integrity were improved with LDL in the extender, as compared to the TCG and TCF. The highest post-thaw integrity of acrosome and plasma membrane and motility were obtained with 9% LDL (w/v). Consequently, the optimum LDL concentration in the extender was 9%. It is also suggested that the concentration of LDL addition is important for the effect on boar sperm protection during freezing and thawing. The percentage of motile spermatozoa was significantly higher after freezing in 9% LDL than in TCG and TCF 54.4% versus 30.4% and 30.1% (p<0.05), respectively. The integrity of acrosome and plasma membrane were also significantly higher at 70.3% and 50.5% respectively with semen frozen in 9% LDL extender compared to TCG at 37.8% and 30.3% and TCF at 36.4% and 29.9%, respectively (p<0.05),. In conclusion, we propose that extender containing LDL extracted from hen egg yolk could be used as a cryoprotective media with a better efficiency than TCG and TCF. LDL improved boar semen quality, allowing better spermatozoa motility, acrosome and plasma membrane integrity after the freeze-thaw process. Furthermore, we found out that the extender with 9% LDL concentration significantly enhanced motility, acrosome and plasma membrane integrity of boar sperm after freezing and thawing.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.2
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• pp.181-189
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• 2008

The aim of this study was to evaluate the effects of $Ca^{2+}$, $HCO_3{^-}$ and BSA on the in vitro capacitation-associated protein tyrosine phosphorylation, hyperactivation and acrosome reaction in guinea pig sperm. Caudal epididymal sperm were incubated in four different groups: modified TALP (Tyrode's albumin lactate pyruvate) or TALP without one of the medium constituents ($Ca^{2+}$, $HCO_3{^-}$ and BSA). After incubation for the required time (0 h, 0.5 h, 1 h, 3 h, 5 h, and 7 h), sperm were removed for further experiment. The capacitation effect was assessed by CTC (Chlortetracycline) staining. Western blotting and indirect immunofluorescence were used to analyze the level and localization of tyrosine phosphorylation. The results showed that guinea pig sperm underwent a time-dependent increase in protein tyrosine phosphorylation during the in vitro capacitation and the percentage of protein tyrosine phosphorylated sperm increased from 36% to 92% from the beginning of incubation to 7 h incubation. Also, there was a shift in the site of phosphotyrosine-specific fluorescence from the head of sperm to both the head and the flagellum. Moreover, an absence of $Ca^{2+}$ or $HCO_3{^-}$ inhibited in vitro hyperactivation and acrosome reaction and decreased the phosphorylation of the proteins throughout the period of in vitro capacitation. However, an absence of BSA could not influence these processes if substituted by polyvinyl alcohol (PVA) in the medium.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.36 no.6
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• pp.641-645
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• 2003

Spermatogenesis, ultrastructure, and sperm morphology of the Pacific oyster (Crassostrea gigas) were investigated with TEM and SEM. C. gigas sperm were primitive consisting of a head midpiece and tail. Sperm size (head and midpiece) was about 1.78 ${\mu}m$. Sperm morphology was similar to a sharp of a small water jar with a rough surface. Sperm had both anterior nuclear fossa (anf) and posterior nuclear fossa (pnf). Acrosome forms had a hat-like shape. The axial rod was projected in front of the acrosome. C. gigas sperm had four large mitochondria in the midpiece.

• Reproductive and Developmental Biology
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• v.38 no.4
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• pp.171-177
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• 2014

L-Carnitine is an antioxidant for the transport of fatty acids in mitochondria and breakdown of lipids for metabolic energy. Some studies have suggested that carnitine improves sperm motility in mammals. The objective of this study was to investigate the effect of L-carnitine on the characteristics in fresh semen of miniature pigs. The collected fresh semen was stored in modena B medium with L-carnitine (0, 1.0, 2.0, and 4.0 mg/ml) for 10 days at $18^{\circ}C$. The semen quality of viability, acrosome reaction and mitochondria integrity was analyzed on 0, 3, 7, and 10 day of semen storage. The percentages of live and dying sperm were not different among treatment groups with different concentrations of L-carnitine during the storage period. In acrosome reaction analysis, when the sperm stored for 7 day, the percentages of live sperm with acrosome reaction were significantly (p<0.05) lower in 1 ($9.0{\pm}0.9%$), 2 ($7.6{\pm}0.2%$) or 4 mg/ml ($7.9{\pm}0.8%$) L-carnitine-treated groups than the control group (0 mg/ml L-carnitine) ($11.12{\pm}0.2%$). However, there were no difference in percentages of live sperm with acrosome reaction for 3 and 10 days of storage with each concentrations of L-carnitine. When sperm was stored for 3 and 10 days, the percentages of live sperm with mitochondria integrity were significantly higher in 2 mg/ml of L-carnitine-treated group than control group (p<0.05). In conclusion, the L-carnitine has a positive effect on acrosome reaction and mitochondria integrity in liquid state of fresh semen in miniature pigs.