To investigate the effect of heparin on sperm capacitation. The acrosome reaction of bovine sperm which were incubated in mTALP containing heparin and the in vitro development of bovine follicular oocytes which were cocultured with heparin-treated sperm were evaluated, and the resutls were as follows : 1. When bovine fresh sperm were incubated in mTALP solution containing 0, 5, 10 and 25$\mu\textrm{g}$/ml heparin for 15 to 840 minutes, there was no significant difference between motilies of heparin-treated sperm and untreated sperm, but the acrosome-reaction rate of heparin-treated sperm was significantly higher than that of untreated sperm. Moreover the acrosome reaction rate of was sperm treated with heparin was significantly increased after incubating for 15 minutes. 2. When fresh sperm were incubated in mTALP solution containing 10$\mu\textrm{g}$/ml heparn for 840 minutes, the motility of sper incubated for 840 minutes was lower than those of sperm incubated for 0, 15, 60 and 120 minutes, but the acrosome-reaction rate of sperm incubated for 840 minutes was higher than those of the others. 3. When frozen-sperm were incubated in mTALP solution containing 10$\mu\textrm{g}$/ml heparin for 120 minutes, the acrosome reaction rate of sperm was significantly increased after incubating for 15 minutes. 4. When fresh sperm treated with heparin were cocultured with bovine follicular oocytes, 16.7 to 23.7% of the oocytes were developed to 2-8 cell stage.
Effects of caffeine, heparin and caffeine-heparin treatments for in vitro capacitation of Korean Native Cattle sperm on acrosorne reaction and viability were studied using the methods of Wells-Awa and Dual stain. The results were summerized as follows: 1. The acrosome reaction of sperm when treated with caffeine after 0 to 4 hrs of preincubation were 11.0~75.7% for Wells-Awa stain, and 14.3~75.55% for Dual stain. True acrosome reaction of sperm for Dual stain was 3.0~29.2%. The viability of sperm was 62. 2~27.2%. 2. The acrosome reaction of sperm when treated with heparin after 0 to 4 hrs of preincubation were 17.0~81.2% for Wells-Awa, and 14.3~75.5% for Dual Stain. True acrosome reaction of sperm for Dual stain was 1.5~26.6%. The viability of sperm was 58.6~35. 8%. 3. The acrosome reaction of sperm when treated with caffeine-heparin after 0 to 4 hrs of preincubation were 13.0~83.2% for Wells Awa, and 11.0~78.5% for Dual stain. True acrosome reaction of for Dual stain was 5.1~26.3%. The viability of sperm was 60.5~30.1%.
This study was carried out ot investigate effect of pH stimulation on acrosome reaction of bovine spermatozoa. The results obtained were as follows : 1. When sperm was sequentially washed with SHP solution of pH 7.4, 7.7 and 7.4 and incubated in mTALP solution of pH 7.4 for 120min, 15, 30, 60 and 120min incubations showed significantly(p<0.05) higher sperm acrosome reaction rate than 0 min. 2. When sperm was sequentially washed with SHP solution of pH 7.4, 8.0 and 7.4 and incubated in mTALP solution of pH 7.4 for 15 minutes, sperm acrosome reaction rate was significantly(p<0.01) increased until 9 min. Incubation, but not increased thereafter. 3. When sperm were separately washed with SHP solutions of pH 7.0, 7.4 and 8.0 and incubated in mTALP solution of pH 7.4 for 9min, sperm acrosome reaction rate was 74.8, 71.8 and 93.4%. pH 8.0 showed signifciantly(p<0.01) higher sperm acrosome reaction rate than pH 7.0 and 7.4. The results suggest that stimulation of sperm with high pH induces sperm crosome reaction.
This study was carried out to investigate the effect of heparin, CSA and PC12 on sperm motility and acrosome reaction in bovine fresh and frozen semen which were washed and incubated in mTALP, and also the effect of heparin-pretreatment on motility and acrosome reaction in mTALP, and also the effect of heparin-pretreatment on motility and acrosome reaction of sperm treated with PC12, and the results obtained were as follows : 1. When fresh sperm was once washed and then incubated for 15 minutes in mTALP containing heparin 1, heparin 2, CSA and PC12, the percent of motile sperm of PC12 was significantly lower than that of control, heparin 1, heparin 2 and CSA. But the percent of acrosomereacted sperm of PC12 was signifciantly higher than that of control, heparin 1, heparin 2, and CSA. 2. When frozen sperm was once washed and then incubated for 15 minutes in mTALP containing heparin 1, heparin 2, CSA and PS12, there was no significant difference in the percent of motile sperm among treatments, but the percent of acrosome-reacted sperm of PC12 was signifciantly higher than that of heparin 2, and there was no significant difference in the percent of acrosome-reacted sperm among control, heparin and CSA. 3. When fresh sperm was twice washed and then incubated for 15 minutes in mTALP containing heparin and PC12, there was no significant differrence in the percent of motile sperm among treatments, but the percent of acrosome-reacted sperm of PC12 was significantly higher than that of control and heparin. When the sperm was incubated for 120 minutes, the percent of motile sperm of PC12 was significantly lower than that of control and heparin, but the percent of acrosome-reacted sperm of PC12 was significantly higher than that of control and heparin. 4. When fresh sperm was twice washed and preincubated in mTALP containing heparin for 0, 15, 120, and 240 minutes, and then incubated with PC12 for 15 minutes, there was no significant difference in the perce수 of motile sperm among treatments, but the percent of acrosome-reacted sperm of 120 and 240 minutes was significantly higher than that of 0 and 15 minutes.
This study investigated the influence of sodium bicarbonate (NaHCO3) and progesterone on acrosome reaction and proportion of polyunsaturated fatty acid (PUFA) composition boar sperm. The sperm were diluted with semen extender and incubated with NaHCO3 and progesterone at 38℃, 5% CO2 for 6 h. Plasma membrane integrity and acrosome reaction were analyzed using SYBR14/propidium iodide (PI) and FITC-PNA/PI doubling staining method, and proportion of PUFA was analyzed using gas chromatography. In results, Plasma membrane integrity was significantly decreased in 50 mM NaHCO3 group and acrosome reaction was significantly increased by over the 100 mM NaHCO3 group compared to control group (p < 0.05). In addition, progesterone significantly increased decreased plasma membrane integrity at 100 mM progesterone and acrosome reaction at over the 5.0 µM progesterone (p < 0.05), but there was no difference among the 5.0 to 100 µM groups. PUFAs were significantly decreased in 100 mM NaHCO3 and 50 µM progesterone treatments compared to control group. In summary NaHCO3 and progesterone induce acrosome reaction and reduce PUFA composition in boar sperm, therefore, the results maybe help to understand basically knowledge for the acrosome reaction and PUFA composition in boar sperm.
This study was designed to analyze boar sperm to compare motility, acrosome morphology, viability and ATP by various preservation methods between Duroc boar A with cold shock resistance sperm and Duroc boar B with cold shock sensitivity sperm. Semen volume, sperm concentration, motility and normal acrosome between Duroc boar A and B did not show any differences within 2 h after collection. There were no differences in sperm motility and normal acrosome between boar A and B at 1 day of preservation at $17^{\circ}C$ and $4^{\circ}C$, respectively. However, sperm motility and normal acrosome from 2 day of preservation at $17^{\circ}C$ and $4^{\circ}C$, respectively, were higher for boar A than boar B. The frozen-thawed sperm motility and normal acrosome were higher for boar A than boar B. The sperm viability and ATP concentration according to storage period of liquid semen at $17^{\circ}C$ and $4^{\circ}C$ were higher for boar A than boar B. Also, the sperm viability and ATP concentration of frozen-thawed semen were higher for boar A than boar B. In conclusion, we found out that the original quality of boar semen with cold shock resistance sperm played an important role.
This study was carried out ot investigate effects of X-537A on hydrogen ion concentration insperm washed solution and sperm acrosome reaction. The results obtained were as follows. 1. When bovine sperm was twice washed with SHP solutions of pH 6.8 and 7.4 and again washed with SHP solution containing 4$\mu$M of X-537A, in case of pH 6.8 the sperm washed with 4$\mu$M of X-537A showed signifciantly(p<0.01) higher hydrogen ion concentration in sperm washed solution than the sperm washed without X-537A. 2. When the sperm was twice washed with SHP solution and then washed with SHP solution containing 4$\mu$M of X-537A, sperm acrosome rection rate was signifciantly(p<0.01) increased from 12min after incubation in the sperm washed without X-537A, but was signifciantly(p<0.01) increased from 8 min after incubation in the sperm washed with 4$\mu$M of X-537A. 3. When the sperm was twice washed with SHP solution and then washed with SHP solution containing 0, 4 and 40$\mu$M of X-537A, and then incubated in m-TALP for 120 min, sperm acrosome reaction rate was significantly(p<0.01) increased from 15 min after incubation in 0, 4 and 40$\mu$m OF X-537A. However at 60 min incubation 40$\mu$M of X-537A showed significantly(p<0.01) higher sperm acrosome reaction rate than 0 and 4$\mu$M and at 120 min incubation 4 and 40$\mu$M of X-537A showed signifciantly(p<0.01) higher acrosome reaction rate than 0$\mu$M of X-537A.
The ultrastructures of germ cells during spermatogenesis and sperm morphology in male Mya arenaria oonogai, which was collected on the coastal waters of Samcheonpo, south coast of Korea, were investigated by transmission electron microscopic observations. In the early stage of the spermatid during spermiogenesis, a few granules and a proacrosomal granule, which is formed by the Golgi complex, appear on the spermatid nucleus, and then it becomes a proacrosomal vesicle. Consequently, it becomes an acrosome by way of the process of acrosome formation. The morphologies of the sperm nucleus type and the acrosome of this species have a curved cylindrical type and cone shape, respectively. The spermatozoon is approximately $48-50{\mu}m$ in length including a curved cylinderical sperm nucleus (about $2.65{\mu}m$ long), an acrosome (about $0.64{\mu}m$ in length) and tail flagellum ($40-45{\mu}m$ long). As some ultrastructural characteristics of the acrosomal vesicle, the peripheral parts of two basal rings show electron opaque part (region), while the apex part of the acrosome shows electron lucent part (region). These charateristics of the sperm belong to the family Myidae or some species of Veneridae in the subclass Heterodonta, unlike a characteristic of the subclass Pteriomorphia showing all part of the acrosome being composed of electron opaque part (region). Therefore, it is easy to distinguish the families or the subclasses by the acrosome structures. Exceptionally, In particular, a cylinder-like nucleus of the sperm is curved (the angle of the nucleus is about $20^{\circ}$), as seen in some species of Veneridae (range from $0^{\circ}-80^{\circ}$). The number of mitochondria in the midpiece of the sperm of this species are four, as one of common characteristics appeared in most species except for a few species in Veneridae in the subclass Heterodonta. Cross-sectioned axoneme of the sperm tail flagellum shows a 9+2 structure: the axoneme of the sperm tail flagellum consists of nine pairs of peripheral microtubules at the periphery and a pair of central doublets at the center.
This study was carried out to investigate the effects of Ca, BSA, heparin, semen storage and individual bull on motility and acrosome reaction of bovine fresh sperm and sperm stored in lactose-egg yolk solution(LES) at 5$^{\circ}C$ for 4hours, and the results obtained were as follows: 1. When sperm was incubated in SCS containing Ca, BSA, Ca + BSA, heparin, heparin + Ca, heparin + BSA, and heparin + Ca + BSA for 15 minutes, there was significant difference in sperm motility among the treatments, especially BSA showed significantly higher sperm motility than the others. Also there was significant difference in sperm acrosome reaction among the treatments, especially BSA and Ca + BSA showed significantly higher sperm acrosome reaction than the others. 2. Bull KNC 1 showed significantly higher sperm motility than KNC 1, HOL 1 and 2 in both fresh and stored semen, however KNC 1 showed significantly lower sperm acrosome reaction than KNC 1, HOL 1 and 2. Therefore, there was significant difference in sperm motility and acrosome reaction among individual bulls. 3. When KNC 1 and KNC 2 sperm were incubated in SCS and SCS + Ca, SCS + BSA, SCS + Ca + BSA, SCS + heparin, SCS + heparin + Ca, SCS + heparin + BSA, and SCS + heparin + Ca + BSA, there was significant difference in sperm motility among individual bulls, especially BSA in KNC 1 and BSA, Ca and Ca + BSA in KNC 2 showedsignificantly higher motility than the others. However, there was significant difference in sperm acrosome reaction among individual bulls, Ca in KNC 1 and Ca + BSA in KNC 2 showed higher acrosome reaction than the others.
Kim, Ki-Sun;Hwang, Kyung-A;Kim, Hyoung-Chin;Nam, Ki-Hoan;Choi, Kyung-Chul
Journal of Embryo Transfer
/
v.26
no.4
/
pp.287-296
/
2011
Mammalian fertilization is a complex cascade process consisting of sperm migration through the female reproductive tract, physiological changes to sperm such as sperm capacitation and acrosome reaction, and sperm-egg interaction in the oviduct in vivo. On the other hand, in vitro fertilization (IVF) is a process by which egg cells are fertilized by sperm outside the body: in vitro. IVF has been used for a variety of purposes in reproductive biotechnology for human and animals. The discovery of sperm capacitation in 1951 promoted the development of IVF technology. In the initial stage of IVF, sperm capacitation in preincubation medium was shown to be essential to fuse with eggs. Besides, sperms should detour some of the in vivo regulations for IVF. This review introduces a general mammalian fertilization process, including sperm capacitation, removal of cumulus matrix, acrosome reaction, and sperm-egg fusion and focuses on the roles of key biochemical molecules, signal mechanisms, and genes involved during IVF and novel results of sperm-oocyte interaction elucidated in various gene-knockout mice models.
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