• Applied Microscopy
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• v.31 no.4
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• pp.333-345
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• 2001

The mature sperms of three species of cephalopods (Octopus minar, Octopus ocellatus, Todarodes pacificus) were observed by electron microscopy. The results obtained are as follows: The sperm lengths of Octopus minor and Octopus ocellatus of octopods are long and they are about $390{\mu}m$ and $125\sim130{\mu}m$, respectively, but the sperm length of Todarodes pacificus is short and about $35{\mu}m$. The sperm of Octopus minor has a helical acrosome and a head bent a little like a banana while Octopus ocellatus of octopod has a twisted acrosome and a long rod-shaped head. A number of horizontal stripes are observed as a periodic structure in their subacrosome cavities and dense plugs are formed in the cavities of their heads. On the other hand, the acrosome of Todarodes pacificus is circular cap-shaped, and its head is long and oval. It is notable that two small cavities were observed in its basal acrosome. Juxtanuclear acrosomal materials of high electron density filled the subacrosomal cavity. In the middle piece of mature sperms of Octopus minor and Octopus ocellatus, the mitochondria form the mitochondrial sleeve, but the numbers of mitochondria differ between the species so that they are $11\sim12$ and $8\sim9$, respectively. Meanwhile, in the middle piece of mature sperms of Todarodes pacificus, the mitochondria are separated from the axoneme, forming a mitochondrial spur in which $10\sim13$ mitochondria and some electron dense materials concentrate. The axoneme of Octopus minor, Octopus ocellatus and Todarodes pacificus are of 9+2 type in common, surrounded by 9 coarse fibres. A number of glycogen were observed only in the axoneme of Todarodes pacificus. The coarse fibres were found as far as the main piece of sperm tail in Octopks minor and Todarodes pacificus, while to the end piece of sperm tail in Octopus ocellatus.

• Development and Reproduction
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• v.5 no.1
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• pp.47-52
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• 2001

Predetermination of sex in livestock of offpring is in great demand and is of critical importance to providing for the most efficient production of the animal ariculture. Such a sexing techlology would also enhance the economy of conventional artificial insemination(AI) and aid the porcine industry. The purpose of this study was to evaluate the efficiency of enriching X-bearing porcine sperm using discontinuous percoll gradients and PCR mefhod. Semen was collected from mature boars of proven fertility center (AI center KimHae). Sperm was leaded on the isotonic discontinuous percoll gradient and then it was centrifuged at 120 ${\times}$ g for 20 minutes. After centrifugation, sperm included in each fraction were recovered (7${\times}$10$^6$ sperms/ml) and then sperm genomic DNA was extractedfor the PCR. SRY gene was used to evaluate the ratio between X and Y sperm in the separated fractions. Ju viro ffrtilization wascarried out by adding the unseparated sperm (control) or separated (experimental poop) to the matured oocytes in TCM-199. Embryos for sex determination were obtained at 2 cell stage and then was used for SRY gene amplification. After centrifugation of discontinuous percoll gradient, the most motile sperm was obtained at 95% fiaction (94.4% ${\pm}$ 5.1%, p < 0.01). The PCR analysis evaluated that 30%, 50% and 65% fractions were Y sperm rich, whereas 80% and 95% fractions were X sperm rich. PCR analysis with each porcineembryo showed that 33.3% of control and 66.7% of experimental group were determined as female embryos. In conclusion, in vitro matured oocytes inseminated with sperms (95% fraction) prepared by percoll gradient centrifugation showed high fertilization rates and female embryos than control sperms.

• Clinical and Experimental Reproductive Medicine
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• v.19 no.2
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• pp.169-174
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• 1992

Previous studies have indicated that immunological factor is responsible for the infertility. We have detected sperm antibodies in ZIFT patients which grouped as fertilization failure (A; n=18) and low fertilization rate (${\leq}50%$)(B; n=20). Patients, however, had normal oocytes and sperms. We collected serum from wives and semen from husbands and donors (fertile sperm), if it was needed. We examined class, binding patterns and amounts of antisperm antibodies(ASA) by direct and indirect immunobead binding assay. In group A, 11 husbands were ASA positive showing 62.2% and 61.1% binding with IgA and IgG, respectively, and two wives were ASA positive showing 70.0% and 71.0% binding with IgA and IgG, respectively. Binding sites were mainly at the head of sperms (84%). In group B, 8 husbands were ASA positive showing 37.5% and 40.0% binding with IgA and IgG, respectively, and two wives were ASA positive showing 41.3% and 42.0% binding with IgA and IgG, respectively. Binding sites were also mainly at the head of sperms (78%). For the treatment of ZIFT patients who had fertilization failure at the first trial, we used albumin fractionation method and dilution method with 30% fetal cord serum (FCS) to reduce the titer of ASA. We used partial zona dissection (P.Z.D.) method for wives who have antisperm antibodies in their serum. According to represented method, we could inhance the fertilization rate to 60.0% by albumin fractionation and 20.0% by P.Z.D., respectively. We concluded that the use of micromanipulation like P.Z.D. or the other sperm processing methods is required to increase a chance of fertilization. This result suggested that it should be a prerequisite to test antisperm antibodies prior to entering assisted reproductive technologies (ART) programs.

• Journal of Embryo Transfer
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• v.29 no.4
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• pp.397-401
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• 2014

The boar sperm has more lipid droplets and specialty of seminal plasma compared with other species, causing difficulties of freezing sperm and decreases for the utilization of frozen semen into the artificial insemination. However, several studies reported significant results for the recovery of sperm motility and reproductive by addition of cryoprotectants and seminal plasma after thawing. This study was designed to investigate the effects of supplementation of trehalose or glycerol in the LEY (lactose and egg yolk in BTS) solution for the conventional freezing and vitrification process. Two boars aged 16 months were used to collect semen for 2 times in a week. The samples were allotted to 3 freezing solutions (LEY + glycerol 10.5% + OEP 1.5%, LEY + trehalose 1M + OEP 1.5%, and sucrose 1.5M + trehalose 1 M + OEP 1.5%) after centrifugation at 800 g for 10 minutes. Semen was equilibrated in freezing solutions for 10 minutes and injected into plastic straws with 2~3 air bubbles to minimize freezing damages. Vitrification was performed to locate sperm in 5 cm above $LN_2$ for 5 minutes, and the conventional freezing was conducted with an automatic freezer. Motility and survival rates were measured by CASA (Computer assisted sperm an alyzing system) and FITC (Fluorescein isothiocyanate), respectively after thawing semen at $50^{\circ}C$ for 12 seconds. The results were analyzed by ANOVA with STATVIEW statistical program. The vitrificatioin solution (LEY + 10.5% glycerol + 1.5% OEP) presented higher motility (20.9%) than other solutions while the solution (LEY + 1M trehalose + 1.5% OEP) showed the lowest (motility : 5.2%). However, survival rates of vitrified sperms detected by FITC showed 1~4% live sperms in almost of dead sperms at all vitrification solutions' groups, but survival rate of freezing solution of LEY + 1M trehalose + 1.5% OEP LEY and LEY + 10.5% glycerol + 1.5% OEP were showed 49%, and 79%, respectively. There were differences (P<0.05) survival rate of conventional freezing in LEY + 10.5% glycerol + 1.5% OEP and LEY + 1M trehalose + 1.5% OEP and the remaining showed no differences. The results suggested that vitrified boar semen was not enough to be utilized for the artificial insemination, but it showed possibility to utilize for ICSI and conventional freezing with glycerol would be useful method for artificial insemination in pig while we choose the outstanding semen against tolerance to freezing damages.

• Journal of the Korean Institute of Intelligent Systems
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• v.17 no.1
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• pp.86-93
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• 2007

A new operation termed competitive generation in the processes of genetic algorithms is proposed for accelerating the optimization speed of genetic algorithms. The competitive generation devised by considering the competition of sperms for fertilization provides a good opportunity for the genetic algorithms to approach global optimum without falling into local optimum. Experimental results with typical problems showed that the genetic algorithms with competitive generation are superior to those without the competitive generation.

• 제어로봇시스템학회:학술대회논문집
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• 1989.10a
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• pp.1135-1139
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• 1989

Three layered neural network was applied for the pattern recognition problem of human spermatozoa in clinical test. The goodness of recognition rate was studied in relation to the number of hidden layer cells and of output layer cells. The proposed method provided better results than conventional template matching technique. Parallel processing of the back propagation learning algorithm was also studied using transputers and its performance was evaluated.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.4
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• pp.405-415
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• 2000

Objective: The present study was undertaken to synthesize a human Y chromosome specific probe and to confirm the usefulness of the probe for fluorescence in situ hybridization (FISH) in various types of human cells. Methods: An approximately 400 bp DNA fragment of the DYZ1 sequences was synthesized by PCR using digoxigenin labeled dUTP (dig-PCR). The fidelity of probe was tested by FISH for cultured and uncultured human lymphocytes, amniocytes, chorionic villus cells, embryos, sperms, and germ cells of seminiferous tubule. Results: The human Y chromosome specific probe hybridized specifically to Y chromosome of the cells that had been tested. This probe assigned to the Yq12 region where the DYZ1 repetitive sequence is concentrated. Conclusion: We have identified a human Y chromosome specific probe that hybridized specifically to the Y chromosome by FISH for various types of uncultured as well as cultured cells. Therefore FISH technique using human Y chromosome specific probe should be useful for clinical application as a diagnostic tool for the detection of human Y chromosome.

• Clinical and Experimental Reproductive Medicine
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• v.18 no.2
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• pp.233-235
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• 1991

A total of 167 patients underwent mascroscopic vasovasostomy by a modified double layer reanastomosis to correct postvasectomy sterility during a 5-year period between 1986 and 1991. We obtained the anatomical patency and pregnancy rates from 61 patients whose follow-ups were completed. There by, we report the following results. 1. Of the 167 patients, the mean age and the average duration of vasobstruction were 34.0 and 4. 4 years respectively. The reasons for ecanalization were desire for more baby in 71.9%. death of children, 24.5% and remarriage in 3.6%. 2. Of the 61 patients with complete follow-up. the anatomical patency and pregnancy rates were 83.6%(51 patients) and 50.8%(31 patients) respectively. 3. For the 36 out of 61 patients whose duration of vasobstruction was less than 5 years, the anatomical patency and pregnancy rates were 88.9%(32 patients) and 58.3%(21 patients) respectively. The rates for the remaining 25 patients whose duration was greater than 5 years were 80.0%(20 patients) and 40.0%(10 patients). 4. Of the 61 patients, 51 exhibited sperms from the proximal vas on microscope during the operation. Their anatomical patency and pregnancy rates were 88.2%(45) and 54.9%(28) respectively. The rates for the remaining 10 patients without any sperms were 60.0%(6) and 30.0%(3). From the above results, we can conclude that macroscopic reanastomoses by modified double layer technique has appreciable success rates that could possibly be compared to the microscopic results.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.1
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• pp.65-72
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• 2001

Objectives: This study was to investigate the fertilization rate after intracytoplasmic sperm injection (ICSI) or partial zona dissection (PZD) of human and hamster sperm into hamster oocyte in in vitro fertilization (IVF). In addition, the possibility of clinical application was evaluated by the comparison of usefulness and difference of these method. Materials and Methods: Hamster immature oocytes were obtained from oviducts superovulated by PMSG and hCG, and hamster sperms were obtained from epididymis. The freezed human sperms were thawed before use. Fertilization were confirmed by two pronuclei, one pronucleus, swollen sperm head or/and two polar bodies at $7{\sim}8$ hour after ICSI or PZD. Results: The fertilization rates after ICSI and PZD of human sperm to hamster oocyte were 3.6%, 64.2%,73.6%, and 55.6% for negative control, positive control, ICSI, and PZD respectively, suggesting that ICSI only showed improved fertilization rate (p<0.01). The fertilization rates after ICSI and PZD of hamster sperm to hamster oocyte were 11.1%, 51.2%, 39.6%, and 72.7% for negative control, positive control, ICSI, and PZD respectively, suggesting that PZD only showed improved fertilization rate (p<0.01). PZD showed significantly higher fertilization rate than ICSI (p<0.05). Conclusions: As for the fertilization rate by ICSI and PZD using hamster oocyte in IVF, ICSI technique was considered to be more useful for human sperm and PZD technique for hamster sperm. Therefore, ICSI technique was considered more appropriate for experimental application using human sperm and hamster oocyte.

• Bulletin of the Korean Chemical Society
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• v.33 no.5
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• pp.1536-1540
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• 2012

Chicken primordial germ cells (cPGCs) are founder germ cells in embryonic stage of development that eventually give rise to sperms or oocytes. Currently cPGCs are only known cells enabling germline transmission in chicken and their cultivation protocols were recently established. Although genome modifications of chickens are now theoretically possible using cPGCs, there are still several hurdles to overcome to practically use cPGCs as mediators for chicken transgenesis. First, efficiency of gene delivery into cPGCs remains low with current methods. Second, there aregene silencing mechanisms against the expression of foreign genes in cPGCs. In this study, we successfully increased the efficiency of gene delivery in cPGCs by taking advantage of the TTAA-specific $piggybac$ transposon system. Moreover, a pipette-type electroporator significantly enhanced transfection efficiency up to 5-fold compared withcuvette-type methods. Taken together, the technological advances in our study will provide practical benefits for the application to fulfill genetic modifications of chicken genome.