• Journal of Embryo Transfer
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• v.16 no.3
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• pp.203-211
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• 2001

This study was carried out to determine sex of porcine embryos produced by in vitro fertilization. Porcine oocyte-cumulus complexes were cultured in BSA-free North Carolina State University (NCSU) 23 medium containing porcine follicular fluid (10%), cystein (0.1 mg/ml) and hormonal supplement (10 IU eCG and 10 IU hCG per ml) for 20~22 hrs. They were then cultured in the same medium but without hormonal supplement for additional 20~22 hrs. After culture, cumulus cells were removed and oocytes were co-incubated for 6 hrs with four different concentrations (5$\times$10$^4$, 2.5$\times$ 10$^{5}$ , 5.0$\times$10$^{5}$ and l0$\times$10$^{5}$ ) of porcine sperm. After fertilization, oocytes were transferred into NCSU 23 with 0.4% BSA medium. The cleavage and blastocyst formation rates were evaluated at 48 and 144 hrs, respectively. In this study, the polymerase chain reaction (PCR) was used to determine the sex of porcine embryos in the stage of blastocyst. The PCR was performed using a set of oligonucleotide primers (5‘-TCATGGACCAGGTAGGGAAT-3', 5’-GAAAGACACGTCCTTGGA GA-3') for 491 bp fragment of porcine male-specific DNA sequence. In the flour different sperm concentration (5$\times$10$^4$, 2.5$\times$10$^{5}$ , 5.0$\times$10$^{5}$ and l0$\times$10$^{5}$ ) for fertilization condition, the cleavage rate was 55.95, 67.88, 60.18 and 47.60%, respectivety, and the development rate of blastocysts was 16.03, 20.40, 21.41 and 12.37%, respectively. At 5.0$\times$10$^4$and 2.5$\times$10$^{5}$ of sperm concentrations per ml cleavage rate and development rate of blastocyst were higher than those of 5.0$\times$10$^4$and l0$\times$10$^{5}$ of sperm concentration (P<0.01). The male of porcine embryos was detected at 491 bp by PCR, and 18 of the 31 porcine blastocysts were the male (58.1%) and the rest 13 were the female(41.9%).

• Korean Journal of Animal Reproduction
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• v.20 no.4
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• pp.395-412
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• 1997

Mammalian eggs are ovulated arrested at meiotic metaphase II until fertilization. Generally in mammals, fertilization results in a series of intracellular calcium oscillations that are mediated by inositol triphosphate (IP$^3$) or cyclic adenosine diphosphoribose (cADPr). The high levels of maturation promotion factor (MPF) within the cell decrease, pronuclei form, the cytoskeleton is reorganized and proteins are post-translationally modified. If all is normal, the newly formed embryo initiates the developmental program specific to that species. Artificial methods of producing these effects in pig oocytes are discussed. One potential mechanism mediated via a signal transduction pathway is present in pig oocytes. Stimulation of this pathway leads to the early events following fertilization, and electrical stimulation leads to apparently normal de v velopment to day 12. Further studies are needed to determine which mechanism(s) the sperm uses to initiate development.

• Animal Systematics, Evolution and Diversity
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• v.12 no.3
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• pp.253-264
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• 1996

The ultrastructure of spermatozoa in urodeles and primitive anurans was examined and compared. The spermatozoa of urodeles are characterized by seven plesiomorphies in subacrosomal cone, endounclear canal. perforatorium, ring, marginal filament, undulating membrane and tail axis. Most primitive anuran spermatozoa have no marginal filament, subacrosomal cone and ring structure with the exception of having the subacrosomal cone in Ascaphus and the ring in Discohlossus as compared with those of urodeles. Persistence of the subacrosomal cone and the ring structure is typical in most urodeles and is further linked with the primitive anurans. Therefore, these characters are regarded as symplesiomorphies in urodeles and primitive anurans. The organization of sperm tail, endounclear canal and perforatorium indicates a close phylogenetic relationship between urodeles and the primitives anurans.

• 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.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.37-37
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• 2001

포유동물의 정자형성과정 (spermatogenesis) 은 유사분열과 감수분열이 동시에 일어나는 매우 복잡하지만, 효율적으로 생식세포를 증식, 분화시키는 시스템이다 정상적인 spermatogenesis가 일어나는 testis에서는 haploid (IN), diploid (2N), 그리고 tetraploid(4N)과 같은 핵형을 갖는 세포들이 일정한 비율로 존재한다. DNA flow cytometry(DNA FCM) 는 세포의 핵형(ploidy)을 신속·정확하게 측정하여, 1N, 2N 그리고 4N에 대한 비율을 예측할 수 있어서, 생식세포를 포함한 다양한 유형의 세포주기를 분석하는데 적용되어져 왔다. 세포주기 분석법 중 이와 같은 FCM이외에, flow cytometer와 static image cytometer를 결합시켜 새롭게 고안된 laser scanning cytometer (LSC)가 있다. 그리고, 이제까지 LSC를 사용한 spermatogenesis에 관한 연구에 대해서는 보고된 바가 없다. 본 실험은 설치류에 있어 각기 다른 발달단계에 있는 정상적인 정소세포를 분리하여 PI (propidium iodide) 로 DNA를 염색한 후, DNA함량을 LSC로 분석하였다. 이것을 FCM에 의한 정소세포의 DNA분석과 비교·검토하였으며, 이 방법을 정상적인 spermatogenesis 가 일어나지 않는 동물시스템에 적용시켰다. 생식세포를 소멸시키기 위해 항암제인 busulfan과 비타민 A를 결핍시켜 이것이 세포주기의 어떤 시점에서 어떻게 작용하여, 생식세포를 소멸시키는지 알아보았다. 위의 실험·분석결과로부터 LSC를 사용한 DNA함량과 핵형의 결정은 FCM과 동일한 수준의 정확성을 제시하였다. busulfan 또는 비타민 A의 결손은 살아있는 세포의 80% 이상이 2N의 핵형에 해당하는 G0/G1 기에 있는 것으로 나타났다. 그리고 1N:2N 및 4N:2N의 핵형비율의 변화를 가져왔다. 이러한 자극은 생식세포주기제어에 관여하며, 생식세포가 증폭하고 분화로 들어가는 단계를 차단, G0/G1 기에서 정체(arrest)되는 것으로 시사된다.

• Korean Journal of Construction Engineering and Management
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• v.12 no.1
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• pp.125-132
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• 2011

Since early 2000s, the importance of CM services has been increasingly recognized and the CM market is continuously growing. As a consequence, the number of CM companies has increased, which means keener market competition. In order to increase their competitiveness, it has been a major task for CM companies to employ and foster competent construction managers who can play a role of efficient project leaders. The objective of the paper is to analyze key characteristics of construction managers' roles during design stages from the perspective of leader's roles including strategic planner, team builder, gatekeeper, expert, champion.

• Development and Reproduction
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• v.10 no.2
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• pp.79-84
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• 2006

We reviewed sex-change patterns in the wrasses Halichoeres poecilopterus, H. tenuispinis, Pteragogus flagellifer, and Pseudolabrus sieboldi inhabiting the coastal waters of Jeju, Korea, based on the sex distribution according to standard length and sex characteristics of the gonads. Halichoeres poecilopterus, H. tenuispinis, Pt. flagellifer, and Ps. sieboldi are protogynous hermaphroditic fish. Histological observations revealed that these wrasses are undelimited type 2 species because testicular tissue(spermatogenesis area) appears in most parts of the gonads during ovary of degenerative stage. Both initial- and terminal-phase males were present in the investigated populations, indicating that Halichoeres poecilopterus, H. tenuispinis, and Ps. sieboldi are of the diandric type. In contrast, Pt. flagellifer is considered a monandric type, because all males in the investigated populations were terminal-phase males produced via sex change from functional females.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.531-539
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• 1996

The lectin-binding patterns in the testis of the sexually matured Jindo dog were investigated to study the distribution of glycoconjugates in the seminiferous tubule under light and transmission electron microscopy. Positive reactions to Wheat germ agglutinin(WGA) and Dolichos biflorus agglutinin (DBA) were observed in the Sertoli cell and in the residual body of spermatid with a stronger reaction in the Sertoli cell to the lectins than in the residual body. Strong reactions to Soybean agglutinin(SBA) and Peanut agglutinin(PNA) were observed in the acrosome vesicles of the Golgi- and cap-phase spermatid, while a moderate reaction was observed in the acrosome-phase, maturation-phase spermatid and the residual body. The acrosome area of the spermatid reacted intensively to Griffonia simplicifolia agglutinin( GS-I) when the cell was in the acrosome-phase and maturation-phase, and the same reaction to the GS-I was observed in the residual body. However, the seminiferous tubule did not react to Ulex europeus agglutinin I(UEA-I). The gold-labelling of the Sertoli cells with DBA resulted in positive reactions of the Sertoli cell column and processes when observed under the electron microscopy, while the Golgi-, cap- and acrosome-phase spermatids reacted positively to SBA in the peripheral low-dense area of the acrosome vesicle of spermatid. Based on these results, we concluded that differences in the lectin-binding pattern of the seminiferous tubules were recognized in the Jindo dog compared to other animals.

• Development and Reproduction
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• v.3 no.1
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• pp.1-13
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• 1999

It is believed that genetic defects make an important contribution to male infertility. Since spermatogenesis is such a complex process, it seems inevitable that many genes are involved in controlling the entire development of germ cells. Genes for infertility, however, are considered to be only those which are defected in the reproduction ability, but normal in other functions. Microdeletions of the Y chromosome have been observed frequently in infertile males. At least two genes, RBM and DAZ, are known to present in the loci where microdeletions occur frequently. A number of autosomal genes were also considered as candidates of infertility genes, based on phenotypes of knockout mice that were deficient of these genes.

• Clinical and Experimental Reproductive Medicine
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• v.22 no.3
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• pp.273-278
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• 1995

Mammalian, including human, spermatozoa undergo morphological and physiological changes during sperm maturation. There were, these changes may affect the fertilization and embryonic development. In this study, we examined the pronucleus formation, pronucleus disappearance and embryonic development in the human oocytes fertilized by intracytoplasmic sperm injection (ICSI). The injected spermatozoa were grouped into ejaculated, epididymal and testicular by the collecting region. Among 363 metaphase II injected oocytes, 287(79.1%) oocytes were normally fertilized and displayed two pronuclei. There were no difference in the fertilization rates and in the pronucleus formation and pronucleus disappearance at 16, 20 and 24 hr after ICSI, among the each spermatozoa group. Also, at 64 hr, the appearance of embryonic development was similar. From these results it can be concluded that there was no difference of maturity among the sperm collected from ejaculated, epididymis and testis in the pronucleus formation and embryonic development. Therefore, testicular spermatozoa are successfully used with ICSI in IVF-ET program.