• Clinical and Experimental Reproductive Medicine
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• v.29 no.1
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• pp.57-66
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• 2002

Objective : This study was designed to investigate the interrelationship and clinical usefulness of sperm morphology by strict criteria (SM), acrosome reaction following ionophore challenge test (ARIC) and sperm penetration assay (SPA) using zona-free hamster ova as prognostic factors in in vitro fertilization. Materials and Methods: Semen samples were provided by 83 patients undergoing IVF. We first evaluated the differences between normal fertilization group and poor fertilization group on three andrologic tests. Secondly, we analyzed the relationship between the three andrologic tests and in vitro fertilization on IVF settings. Finally, we evaluated the effectiveness of the three andrologic tests as the prognostic indicators for fertilizing ability. Results: The fertilization rate of all men in the poor fertilization group was less than 30%; but there was no evidence that this poor fertilization was due to oocyte defects. The results of three andrologic tests were significatly higher in normal fertilization group. Fertilization rate (%) in vitro was highly correlated (p<0.001) with % normal sperm by SM, ARIC value (%), and SPA result. By using Receiver-Operator-Characteristic curve (ROC), we evaluated the effectiveness of these three tests. The sensitivity and specificity of SM, ARIC test and SPA in predicting fertilization potential in IVF setting were 76% and 75%, 84% and 90%, and 76% and 95%, respectively. Conclusion: Our data suggest that the three andrologic tests can be reliable tools as prognostic factors of sperm fertilizing ability. Among these test, ARIC test and SPA gave more accurate information on fertilizing capacity. ARIC test was shown to have a predictive value for fertilizing ability comparable to that of SPA that appears to be a simple and cost-effective addition to current andrology laboratory. Combined application of these three tests may give more information on predicting sperm fertilizing capacity.

• Reproductive and Developmental Biology
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• v.31 no.3
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• pp.175-179
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• 2007

The objective of this study was to investigate the effect of thawing temperature on the sperm viability and acrosomal morphology for semen storage of miniature pig by the 0.5ml straw method. In this present study, sperm viability (SYBR-14/PI staining), membrane integrity (Hypoosmotic Swelling Test), acrosome intactness, intensity and capacitation status (chlorotetracycline staining) in frozen miniature pig sperm were evaluated after thawing at 37, 50 and $70^{\circ}C$ for 5, 10 and 45 sec, respectively. Interestingly, the results indicated that sperm thawed at $70^{\circ}C$ for 5 sec significantly (p<0.05) increased sperm viability, but lower the percentage of AR (acrosome reacted spermatozoa) pattern compared to sperm thawed at $37^{\circ}C$ for 45 sec and $50^{\circ}C$ for 10 sec. In terms of thawing condition, high temperature for a short time using the 0.5ml straw was improved cryosurvival of miniature pig semen. Therefore, appropriate thawing method for cryopreservation of miniature pig is required for increasing post-thawing viability.

• Journal of Animal Science and Technology
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• v.56 no.7
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• pp.26.1-26.10
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• 2014

Background: Endocrine disruptors are exogenous substance, interfere with the endocrine system, and disrupt hormonal functions. However, the effect of endocrine disruptors in different species has not yet been elucidated. Therefore, we investigated the possible effects of $17{\beta}$-estradiol (E2), progesterone (P4), genistein (GEN) and 4-tert-octylphenol (OP), on capacitation and the acrosome reaction in bovine, mouse, and porcine spermatozoa. In this in vitro trial, spermatozoa were incubated with $0.001-100{\mu}M$ of each chemical either 15 or 30 min and then assessed capacitation status using chlortetracycline staining. Results: E2 significantly increased capacitation and the acrosome reaction after 30 min, while the acrosome reaction after 15 min incubation in mouse spermatozoa. Simultaneously, capacitation and the acrosome reaction were induced after 15 and 30 min incubation in porcine spermatozoa, respectively. Capacitation was increased in porcine spermatozoa after 15 min incubation at the lowest concentration, while the acrosome reaction was increased in mouse spermatozoa after 30 min (P < 0.05). E2 significantly increased the acrosome reaction in porcine spermatozoa, but only at the highest concentration examined (P < 0.05). P4 significantly increased the acrosome reaction in bovine and mouse spermatozoa treated for 15 min (P < 0.05). The same treatment significantly increased capacitation in porcine spermatozoa (P < 0.05). P4 significantly increased capacitation in mouse spermatozoa treated for 30 min (P < 0.05). GEN significantly increased the acrosome reaction in porcine spermatozoa treated for 15 and 30 min and in mouse spermatozoa treated for 30 min (P < 0.05). OP significantly increased the acrosome reaction in mouse spermatozoa after 15 min (P < 0.05). Besides, when spermatozoa were incubated for 30 min, capacitation and the acrosome reaction were higher than 15 min incubation in E2 or GEN. Furthermore, the responsiveness of bovine, mouse and porcine spermatozoa to each chemical differed. Conclusions: In conclusion, all chemicals studied effectively increased capacitation and the acrosome reaction in bovine, mouse, and porcine spermatozoa. Also we found that both E2 and P4 were more potent than environmental estrogens in altering sperm function. Porcine and mouse spermatozoa were more responsive than bovine spermatozoa.

• Applied Microscopy
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• v.28 no.2
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• pp.193-205
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• 1998

The Urechis unicinctus sperm and spermatogenic cells prepared from the testis are investigated to identify $\alpha-tubulin$ of axoneme microtubules using mouse monoclonal $anti-\alpha-tubulin$ as the first Ab and Gold(10nm) conjugated goat anti-mouse IgG as the Ab marker. The Ag-Ab reaction analyzed excellently the localization of $\alpha-tubulin$ and the gold particles incorporated with the proximal and distal centrioles, manchette microtubules, and flagellum. The gold particles can be also observed in the spermatogenic cells while the cells are still in sperm ball which is composed of a somatic cell and spermatogenic cells. The sperm ball is the functional unit of sperm production in U unicinctus testis. The spermatids are developed from the spermatogenic cells in the sperm ball and released into the testis cavity through a cortical cytoplasmic opening. The spermatid architectures are similar with the mature sperm of the testis cavity in aspects of shape of discoid acrosome, degree of nuclear condensation and ring type of mitochondrion. However, the distal centriole connecting with the flagella can be observed from the mature sperm while the both proximal and distal centrioles reveal only in the spermatids. The proximal centriole is directly connected with nuclear outer membrane during the stage of nuclear condensation and oriented perpendicularly to the distal centriole whose axis coinciding with the longitudinal axis of the spermatozoon. There are indications that the distal centriole is intimately associated with the polymerization of the flagellum. The manchette microtubules appear during spermatid development but the mature sperm have round head and no conspicuous middle piece.

• Development and Reproduction
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• v.11 no.2
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• pp.97-104
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• 2007

This study describes the spermatogenesis and sperm ultrastructure of the spiny top shell, Batillus cornutus using light and electron microscopy. The spiny top shells were collected by divers in the coastal water of Wandogun, Cheollanamdo, Korea(N $34^{\circ}13'$, E $126^{\circ}47'$) at May 2003. Spiny top shells of $60.0{\sim}69.9\;mm$ in shell height were used in this study. The testis comprises many spermatogenic follicles which contains germ cells in different developmental stages. The primary spermatocytes in the pachytene stage were characterized by synaptonemal complexes. The early spermatids were characterized by appearance of Golgi complex, increased karyoplasmic electron density and tubular mitochondria. In early spermatid the mass of proacrosomal granules consists of numerous heterogeneous granules with high electron density. From the mid-stage of spermiogenesis the well-developed mitochondria aggregate posterior to the nucleus, and surround the proximal and distal centrioles. In this stage, proacrosomal granules are condensed and form a acrosome with thin envelope. During the late spermiogenesis, the acrosome begins to elongate and then became conical. The sperm consists of head, mid-piece and tail. The head comprises a round nucleus and a conical acrosome. Acrosomal rod of microfibrous is observed between nucleus and acrosome. Five mitochondria observed in mid-piece. And tail has the typical "9+2" microtubular system originates from the centrioles.

• Journal of Veterinary Clinics
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• v.8 no.2
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• pp.183-190
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• 1991

Pellet and straw methods in canine semen freezing are compared with respect to motility, viability and acrosome demage of sperm during each of the two major processing steps, to prior-freezing and to frozen-thawing. Senen was extended with a tris-buffered egg yolk contained 4% glycero1 Pellet freezing in the hole of dry ice and straw freezing on the surface of liquid nitrogen were carried out, respectively. The frozen semen 10 days after storage in liquid nitrogen container. wao thawed. In the comparison of two freezing methods, the straw freezing method with 42.7% in motility. 49.2% in viability and 0.186 acrosome score after thawing seems to be superior to the pellet freezing method with 31.2%, 34.5% and 0.314%, respectively. Sperm motility of processing step to frozen-thawing against decrease rate 12.67% to Prior freezing appeared of 33.84% and 49.37% in straw and pellet freezing and increase of 0.02 in acrsomal score to prior freezing appeared of 0.08 and 0.21 in straw and pellet freezing method to frozen-thawing

• Applied Microscopy
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• v.26 no.3
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• pp.267-275
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• 1996

The internal ultrastructural changes of germ cells and external morphology of spermatozoon during the spermatogenesis in the rockfish, Sebastes inermis were studied using transmission and scanning electron microscope. The testis is seminiferous tubule type in internal structure. Seminiferous tubule consist of many cyst which contain numerous germ cells in same developmental stage. Spermatogonium contained a large nucleus with single nucleolus in interphase. Primary spermatocyte identified by the presence of synaptonemal complex in nucleus and the contained a number of mitochondria, endoplasmic reticula and Golgi bodies in cytoplasm. The nucleoplasm of secondary spermatocyte was more concentrated than that of the previous phase. Spermatids were more condensed in nucleus and cytoplasm, and show the long-spherical shape. In the cytoplasm of spermatid mitochondria located to lower portion of the nucleus and Golgi bodies located to upper portion, but proacrosomal granule is not appeared. The spermatozoon consist of the head and tail. No acrosome could be found in the head. The cytoplasmic collar of posterior part in sperm head contained mitochondria which surrounded axial filament. The well developed axonemal lateral fins were identified in sperm flagellum, and the axial filament of the flagellum consist of nine pairs of peripheral microtubules and one pair of central microtubules.

• Reproductive and Developmental Biology
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• v.29 no.1
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• pp.25-30
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• 2005

This study was carried out to investigate the effects of semen characteristics, frozen-thawed sperm viability and serum FSH, LH, estradiol-17β and testosterone concentrations between breeds and among seasons in boars. In all seasons, Yorkshire boars produced higher semen volume compared with Duroc boars, whereas sperm concentration did not differ significantly between Duroc and Yorkshire boars. Semen volume in spring was higher compared with summer, autumn and winter in both Duroc and Yorkshire boars, but sperm concentration did not differ significantly among seasons. Sperm motility and normal acrosome rate of frozen-thawed sperm produced in spring were higher than those in summer, autumn and winter in both Duroc and Yorkshire boars. Sperm motility of frozen-thawed sperm in Yorkshire boars was higher than that in Duroc boars regardless of seasons. However, normal acrosome rate did not differ significantly between Duroc and Yorkshire boars. Serum FSH concentration in Yorkshire boars was lower than that in Duroc boars in all seasons. However, there were no significant differences on serum FSH concentration of Duroc and Yorkshire boars among seasons. Serum LH and estradiol-17β concentrations did not differ significantly between Duroc and Yorkshire boars. Also, there were no significant differences in serum LH and estradiol-17β concentrations of Duroc and Yorkshire boars among seasons. Serum testosterone concentration in Yorkshire boars was higher than that in Duroc boars in all seasons. In both breeds, serum testosterone concentrations were higher in spring than in summer, autumn and winter. In conclusion, when serum FSH concentrations were low, semen volumes were high, and when serum testosterone concentrations were high, sperm motility and normal acrosome rate of frozen-thawed sperm were high.

• Korean Journal of Animal Reproduction
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• v.23 no.3
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• pp.191-203
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• 1999

The objectives of this study were to investigate 1) the effects of Selenium(Se), Vitamin E (Vit. E) or recombinant Bovine Somatotropin(rBST) administration on fresh and frozen/thawed semen characteristics and 2) the effect of taurine on frozen/thawed semen characteristics in Hanwoo sires Hanwoo sires were randomly assigned to five groups (1. control, 2. rBST, 0.09mg/kg body weight (BW), 3. Vito E 1,500IU/kg BW, 4. Se 0.l mg/kg BW, 5. Vit. E 1,500IU plus Se 0.1 mg/kg BW). The administration of Se, Vit. E and rBST for each experimental group were given 6 times at 15 days interval by intramuscular injection. The administration of Se, Vit. E or rBST in Hanwoo sires didn't affect semen volume and pH values, but sperm viability was significantly increased comparing to the control group. Also, frozen/thawed semen analysis showed that the sperm viability increased, but any other effects were not found in total sperm :lumber, motility and abnormality among treatments. The addition of taurine in semen freezing extender had a beneficial effects on frozen/thawecl semen characteristics in all groups. The administrations of rBST, Vit. E and Se did not affect the sperm capacitation and acrosome reaction, either the ratio of F pattern(uncapacitated and acrosome intact sperm) or AR pattern(capacitated and acrosome-reacted sperm), but the ratio of B patten(capacitated and acrosome intact sperm) of treatment groups was significantly higher than that of control group, These results indicated that the viability, motility and quality of semen in Hanwoo sires were slightly increased by the injection of rBST, Vit. E and Se, and the addition of taurine in semen freezing extender were also increased the semen characteristics after thawing.

• Journal of Embryo Transfer
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• v.20 no.1
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• pp.43-48
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• 2005

This experiment was carried out to investigate the extender, cooling rate and concentration of glycerol for freezing of boar semen. The result obtained were summarized as follows: 1. Optimal cooling rate was $0.17\~022^{\circ}C/min$ from 25 to $5^{\circ}C$ in LEY extender on the viability and normal acrosome after thawed. 2. The LEY extender was effective in protecting frozen boar semen from cold shock among the extenders(p<0.001, respectively). 3. The sperm viability and normal acrosome rates after thawing was showed greater in the 3 or $4\%$ of glycerol concentration than $2\%$ in LEY extender. 4. Viability of sperm was higher when both 15mM of fructose and 3 or $4\%$ glycerol were added to the LEY extender compared with other concentrations of fructose and glycerol were added it(p<0.001).