• 한국패류학회지
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• 제22권1호
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• pp.1-11
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• 2006

The gonadosomatic index (GSI), oogenesis and reproductive cycle in female Neptunea (Barbitonia) afhritica cumingii were investigated by light and electron microscope observations. In the early vitellogenic oocyte, the Golgi complex and mitochondria were involved in the formation of glycogen, lipid droplets and yolk granules. Late in the vitellogenic oocytes, the rough endoplasmic reticulum and multivesicular bodies were involved in the formation of proteid yolk granules in cytoplasm. In particular, compared with the results of other gastropods, it showed a different result that appearances of cortical granules at the cortical layer and microvilli on the vitelline envelope, which is associated with heterosynthetic vitellogenesis, were not observed in vitellogenic oocytes during oogenesis. A mature yolk granule was composed of three components: main body (central core), superficial layer, and the limiting membrane. Monthly changes in the gonadosomatic index in females studied in 2004 and 2005 were closely associated with ovarian developmental phases. Spawning occurred between May and August in 2004 and 2005 and the main spawning occurred between June and July when the seawater temperature rose to approximately $18-23^{\circ}C$. The female reproductive cycle can be classified into five successive stages: early active stage (September to October), late active stage (November to February), ripe stage (February to June), partially spawned stage (May to August), and recovery stage (June to August).

• Clinical and Experimental Reproductive Medicine
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• 제45권4호
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• pp.189-194
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• 2018

Objective: The aim of this study was to compare the rate of maturation, fertilization, and embryo development of in vitro-matured human oocytes derived from pregnant and non-pregnant women. Methods: Immature oocytes were obtained by needle aspiration from 49 pregnant women (group 1) who underwent a cesarean section at term and 77 non-pregnant women (group 2) who underwent a gynecological operation during the same period (8 months). Healthy immature oocytes (530 in group 1 and 539 in group 2) were cultured and assessed for maturation 36 hours later. Mature oocytes were inseminated by intracytoplasmic sperm injection and cultured up to 144 hours. Results: The percentage of degenerated oocytes was significantly higher (12.1% vs. 6.3%; p<0.001) in group 1 than in group 2. There was no significant difference in the maturation rate (66.8% vs. 68.1%; p=0.698), fertilization rate (66.7% vs. 67.6%; p=0.857), or the rate of formation of good-quality blastocysts (46.2% vs. 47.2%; p=0.898) in oocytes obtained from pregnant and non-pregnant women. Conclusion: The developmental competence of immature oocytes did not differ between pregnant and non-pregnant women.

• 한국수정란이식학회지
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• 제19권2호
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• pp.81-87
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• 2004

본 연구는 재래산양에 있어서 과배란 처리시 공란 산양의 반복사용에 따른 oocyte의 회수방법과 회수란의 질적 개선방법을 확립하기 위하여 실시하였으며, 성선자극호르몬의 1회 처리와 반복처리, 호르몬제의 종류에 따른 oocyte의 회수율과 등급을 조사하였다. 공시동물은 체중 15~25kg 전${\cdot}$후의 성숙한 미경산 재래산양으로서 발정 동기화를 위하여 CIDR를 10일간 질내에 삽입하고 과배란 처리는 70 mg의 FSH 와 1,000 IU의 PMSG를 투여하여 과배란을 유기하였다. ${PGF_{2\alpha}$ 는 8일째에 투여하여 과배란을 유기하였으며, CIDR는 10일째 제거와 동시에 hCG 400 IU를 투여하였다. 난자의 회수는 hCG 투여 후 35~40시간째에 외과적인 방법으로 실시하였으며, 회수한 난자는 난구세포와 세포질의 부착정도에 따라 4등급으로 분류하여 회수율을 조사하였다. FSH와 PMSG를 투여하여 공란 산양으로 처음 사용하였을 때 49두에서 324(6.63${\pm}$0.63) 개와 27두에서 305(11.30${\pm}$1.29)개의 배란점을 확인하였으며, 이 중 43.8%(142/324)와 69.2%(211/305)가 회수되어 두당 회수율은 2.92${\pm}$0.52 개와 7.81${\pm}$1.14개로서 FSH와 PMSG간에 전체 회수율과 두당 회수율은 PMSG 처리구가 유의적(P<0.05)으로 높았다. 반복 사용하였을 때 FSH와 PMSG 처리구의 두당 배란수는 2.88${\pm}$0.69 개와 7.56${\pm}$1.11 개였으며, 회수율은 각각 17.4%(4/23) 및 44.9%(61/136) 로 PMSG 처리구가 유의적(P<0.05)으로 높았다. 두당 회수율은 0.50${\pm}$0.27개 및 3.39${\pm}$0.86개로서 PMSG 처리구가 유의적으로 높았다. 회수한 난포란의 1 및 2등급에 있어서 공란 산양으로 처음 사용하였을 때 FSH 투여구는 각각 20.6%(GI) 및 14.9%(GII)였으며, PMSG 투여구는 12.9%(GI) 및 14.1%(GII)로 나타났다. 반복 사용하였을 때, 회수한 난포란의 등급은 FSH 투여구는 각각 31.3%(GI) 및 28.1%(GII)였으며, PMSG 투여구는 36.4%(GI), 5.1%(GII)로 나타났다. 이상의 결과로 볼 때 배란된 난자의 회수율은 PMSG 가 FSH보다 높았으며, 난포내 난자의 회수율은 투여 호르몬 및 반복사용 여부에 따른 차이는 없었다.

• 한국발생생물학회지:발생과생식
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• 제6권2호
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• pp.97-104
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• 2002

대부분의 포유동물에서 수란관내로 배란된 난자는 정자에 의해 수정이 된 후 개체발생을 시작한다. 그러나 수정이 되지 못한 난자들은 난구세포와 함께 수란관내에서 퇴화하여 제거되는데, 그 기작에 대해서는 구체적으로 알려져 있지 않다. 따라서 본 연구는 포유동물의 수란관내 물질이 난자-난구 복합체에 미치는 영향을 알아보고자 사람의 난포액과 소의 수란관 조직 추출액을 생쥐의 난자-난구 복합체에 처리하고 난자의 생존율 및 난구세포의 세포자연사(apoptosis)를 조사하였다. 수란관 조직 추출액을 처리한 미성숙난자는 난포액만을 처리한 난자와 비교하여 난자의 성숙률에 차이를 보이지 않았다. 그러나 난구세포에서 난포액만을 처리한 경우 확장을 일으키면서 배양접시 바닥에 붙어서 배양 후 72시간까지 분열 증식을 계속하는 것이 관찰된 반면, 수란관 조직 추출액을 처리한 난구세포는 확장이 억제되며 72시간 배양 후 모두 죽었다. 한편 난포액과 수란관 조직 추출액을 함께 처리한 난구세포는 배양 후 24시간에 화장을 일으켰으나 72시간 후에는 수란관 조직 추출액 만을 처리했을 때 와 마찬가지로 모든 난구세포가 죽었다. 이러한 난구세포의 죽음이 세포자연사에 의한 것인지를 알아보기 위하여 DAPI로 핵을 염색한 후 관찰한 결과 세포자연사의 특징인 응축되고 분절화된 핵을 관찰 할 수 있었고, 특히 수란관 조직 추출액을 처리한 난구세포에서 많이 관찰되었다. 또한 TUNEL 방법으로 세포자연사를 확인한 결과 응축되고 분절화된 핵을 가진 난구세포에서 염색되는 것을 확인하였다. 본 연구의 결과들을 종합해 볼 때 소의 수란관 조직 추출액에 의한 생쥐 난구세포의 죽음은 세포자연사에 의한 것으로 판단되며, 이로 미루어 수란관 조직세포에는 난구세포의 세포자연사를 유도할 수 있는 물질을 포함하고 있는 것으로 사료된다.or teacher educators to re-conceptualize teacher education in Korea. 것이 필요하다.량은 264,000$m^2$, 79,200$m^3$이였으며, 우려기준의 40% 농도 이상의 경우 복원 면적과 물량은 779,000$m^2$, 233,700㎥, 배경치 농도 이상의 복원 목표 설정의 경우에는 각각 969,200$m^2$, 290,760$m^3$ 이였다. 토양오염 우려기준 농도의 40%를 복원 목표로 하는 경우와 배경치 농도를 복원 목표로 하는 경우, 복원 물량은 우려기준 농도를 복원 목표로 하는 경우의 3.0배와 3.7배정도 증가하는 것으로 나타나 복원 목표치 설정시 우려기준 농도의 40%와 배경농도의 차이에 다른 복원 물량의 차이는 적어서, 복원 목표 설정시 배경치 농도로 복원계획을 세우는 것이 가장 바람직한 것으로 판단되었다.amma}$D)안정동위원소값이 광화I시기에는 각각 11∼9.${\textperthansand}$, -92∼-86${\textperthansand}$, 광화 II시기에는 각각 0.3${\textperthansand}$(${\gamma}^{18}O_{H2O}$),-93${\textperthansand}$({\gamma}$D)이며, 리본-호상구조를 보이는 것으로 보아 대봉광상의 광화유체에 대한 기원과 진화과정을 두 가지로 생각할 수 있다. 1) 마그마유체로부터 광화작용이 진행됨에 따라 계속적인 순환수의 혼입이 있었으며 2) 조기 마그마${\pm}$변성유체에서 유체압력의 차에

• 한국어류학회지
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• 제26권3호
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• pp.179-184
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• 2014

잉어과(Pisces, Cyprinidae)에 속하는 멸종위기 어류인 돌상어 Gobiobotia brevibarba의 난막미세구조 및 난자형성과정을 광학현미경과 주사전자현미경을 이용하여 관찰하였다. 2014년 5월의 돌상어 난소 내에는 다양한 발생단계의 생식세포들이 관찰되었다. 난모세포들의 상대면적은 성숙란(74.5%), 난황물질이 형성되는 난황구기(16.6%), 난황포기(6.6%), 그리고 미숙한 발달단계인 주변인기(2.3%) 순으로 구성되어 있었다. 주변인기 단계의 세포질은 hematoxylin에 강하게 염색되며, 핵막의 안쪽에는 다수의 인들이 존재하였다. 난황형성물질인 난황포기와 초기 난황구기 단계에서는 여포층과 방사대가 뚜렷하게 형성되기 시작되었으며, 방사대 위에는 미세융모의 난막구조가 더욱 뚜렷해졌다. 난황구기 및 성숙란의 단계에서는 난황구 일부가 융합되면서 강한 호산성을 보였다. 또한 성숙란의 표면에는 깔때기 모양의 난문이 관찰되었으며, $2{\sim}3{\mu}m$ 길이의 미세융모가 일정하게 분포하였다.

• 한국양식학회지
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• 제21권2호
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• pp.96-101
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• 2008

능성어 암컷의 난모세포 발달에 따른 성숙 및 배란유도를 위하여 제주도 연안에서 채집하여 사육중인 능성어 51마리(전장 $69.1{\pm}1.0cm$, 체중 $5.8{\pm}0.3kg$)를 대상으로 암컷의 생식소 발달을 cannulation으로 조사하였다. 난황구기 단계 이상 난모세포(난경 $300{\sim}500{\mu}m$)를 가진 어미를 대상으로 HCG 500 IU/kg BW로 주사하였다. HCG 처리 후 난경변화는 24시간째에 $300{\sim}700{\mu}m$, 48시간째에는 $800{\mu}m$ 이상이 $91.3{\sim}98.8%(95.1{\pm}3.7%)$이었다. HCG 처리 실험구에서는 실험어 8마리 중 7마리가 배란되었고, 총 배란된 알 2,480 mL이었으며, 이중 부상란은 1,360 mL이었다. 수정률과 부화율은 각각 $56.2{\sim}94.9%(77.8{\pm}2.9%),\;70.0{\sim}97.7%(92.2{\pm}2.1%)$이었다. 이들 결과들은 HCG 500 IU/kg BW 농도로 처리했을 때 난황구기 난모세포를 가진 능성어 암컷의 성숙 및 배란에 효과가 있을 것으로 사료된다.

• Clinical and Experimental Reproductive Medicine
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• 제27권1호
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• pp.23-29
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• 2000

Objective: Zona pellucida (ZP) has been thought to be the barrier of egg to sperm penetration before and after fertilization. The phenomenon of ZP hardening has been considered as a post-fertilization event until now, and it is generally accepted that it is caused by the secretory products of cortical granules released during the cortical reaction. Hardening of ZP could occur "spontaneously" in mammalian oocytes in standard culture conditions, and that it is probably not a consequence of cortical reaction. The purpose of our study was to investigate the effect of human amniotic fluid (HAF) on nuclear maturation (NM) and fertilization ability of mouse immature oocytes. Methods: HAF was obtained from patients undergoing amniocentesis at $16{\sim}20$ weeks of gestation. HAF from five to ten patients was centrifuged and the supernatants was pooled. Cumulusenclosed mouse immature oocytes were incubated in the medium containing HAF, and examined to confirm NM and fertilization. Female ICR mice (about 3 weeks old) were stimulated with 7.5 IU PMSG. Immature oocytes were isolated at $48{\sim}52$ hrs post PMSG injection and cultured in TCM-199 supplemented with 20% HAF for 18 hrs. FBS was used as a control for the examination. Matured oocytes (MII) were fertilized with sperms collected from the epididymis of male mice (over 10 weeks old). Fertilization was in conducted T6 medium containing 15 mg/ml BSA, and confirmed at 6 hrs post-insemination. Fertilization rate was assessed in zona-intact or zona-free oocytes (denuded by trypsin). Evaluation of NM and fertilization was carried out by rapid staining method. ZP hardening was evaluated by incubating cumulus cell-free mature oocytes in 0.001% chymotrypsin at $37^{\circ}C$ for 10 min. Results: There was no significant difference between the effects of HAF (86.6%) and FBS (87.7%) supplements on NM of immature oocytes. When maturation medium was supplemented with HAF, total fertilization rates (7%) were significantly lower (p<0.01) than that of FBS (85.1%). In HAF group, fertilization rate was increased (p<0.01) in zona-free oocytes (7% versus 100%). The resistance of mouse oocyte ZP to digestion by chymotrypsin after maturation in vitro was significantly higher (p<0.01) in HAF group (86.7%) than in FBS (6.7%). To culture oocytes in FBS were very effective in preventing ZP hardening. However cultured oocytes in HAF showed high rate of ZP hardening (p<0.01). Conclusions: These results suggest that HAF can be used as a supplement for the NM of mouse immature oocytes in vitro. However, HAF induces spontaneous hardening of ZP of mouse immaure oocytes during maturation in vitro.

• 한국패류학회지
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• 제20권1호
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• pp.55-63
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• 2004

Gonadosomatic index, condition index and reproductive cycle with the gonadal development of the female Octopus ocellatus were investigated by histological observations and morphometric data, from January to December, 2000. And changes in biochemical components of the ovary and the trunk tissues including the digestive organ associated with gonadal development were studied by biochemical analysis from January to October, 2001. The specimens were collected at the coastal waters of Buan, Jeollabuk-do, Korea, from January 2000 to October 2001. O. ocellatus is a dioecious organism. The gonad of O. ocellatus locates medially in posterior region of the body. Morphology of the ovary shows round and oval in shape, the average diameter and external colour of ripe ovary was 32 mm and semitransparent light brown in colour. As the ovary was getting mature, transparent elongated eggs covered with chorion were present in the ovarian cavity. Monthly changes in the gonadosomatic index (GSI) showed a similar pattern with those of the condition index. The GSI and condition index began to increase in March and reached the maximum in April. And then, their values decreased from May and reached the minimum in September. Reproductive cycle of O. ocellatus can be categorized into five successive stages: early developing stage (September to December), late developing stage (November to March), ripe stage (March to May), partially spawned stage (April to June), and degenerative/resting stage (June to October). Follicle cells attached to an oocyte were involved in vitellogenesis in the cytoplasm of the vitellogeneic oocyte and formation of chorion (secondary egg membrane) of the ovarian eggs. Spawning occurred between April and June. The spawning period was once a year and the peak took place between May and June. This species belongs to semelparity. According to changes in biochemical contents of the ovary and the digestive organ, monthly variations of moisture, total protein, total lipid and glycogen contents (%) in the ovary showed a negative correlationship with those of the trunk tissues including the digestive organ. Accordingly, it is assumed that the ovary only may be received nutrient supply (total lipid content) for gonadal development from the trunk tissues including the digestive organ (r = -0.55, p < 0.05).

• 한국패류학회지
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• 제27권1호
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• pp.43-53
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• 2011

The gametogenic cycle and the spawning season in female and male Cyclina sinensis were investigated by quantitative statistical analysis using an image analyzer system, and the biological minimum size (the size at 50% of sexual maturity) was calculated by combination of quantitative data by size and von Bertalanffy's equation. Compared the gametogenic cycle by quantitative statistical analysis with the previous qualitative results in female and male C. sinensis, monthly changes in female and male gametogenic cycles calculated by quantitative statistical analysis showed similar patterns to the gonadal stages in female and male reproductive cycles by qualitative histological analysis. Comparisons of monthly changes in the portions (%) of each area to eight kinds of areas by quantitative statistical analysis in the gonads in female and male C. sinensis are as follows. Monthly changes in the portions (%) of the ovary areas to total tissue areas in females and also monthly changes in the portions of the testis areas to total tissue areas in males increased in March and reached the maximum in May, and then showed a rapid decrease from June to October. Monthly changes in the portions (%) of oocyte areas to ovarian tissue areas in females and also monthly changes in the portions of the areas of the spermatogenic stages to testis areas in males began to increase in March and reached the maximum in June in females and males, and then rapidly dropped from July to October in females and males when spawnig occurred. From these data, it is apparent that the number of spawning seasons in female and male C. sinensis occurred once per year, from July to October. Monthly changes in the number of the oocytes per mm2 and in the mean diameter of the oocyte in captured image which were calculated for each female slide showed a maximum in May and reached the minimum from December to February. Therefore, C. sinensis in both sexes showed a unimodal gametogenic cycle during the year. The percentage of sexual maturity of female and male clams ranging from 25.1 to 30.0 mm in length was over 50% and 100% for clams over 40.1 mm length. In this study, the biological minimum size (sexually mature shell lengths at 50% of sexual maturity) in females and males were 26.85 and 26.28 mm, respectively.

• Clinical and Experimental Reproductive Medicine
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• 제26권3호
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• pp.419-432
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• 1999

Objective: To establish the evaluation system of the quality of oocytes on the basis of the incidence of cumulus cells apoptosis, to investigate the relationships beween the incidence of cumulus cells and the outcomes of IVF-ET. Method: Thirth-four cycles undergoing controlled ovarian hyperstimulation for IVF-ET with tubal infertility (23 cycles) or unexplained infertility (11 cycles) were included in this study. Cumulus cell masses surrounding mature oocyte and co-culture of embryos with autologous cumulus cells during IVF-ET process. The incidence of apoptosis in cumulus cells was assessed by apoptosis detection kit fluorescein. The effect of co-culture using cumulus cells and the incidence of cumulus cells apoptosis. Results: The results were as follows: 1. The incidence of apoptosis in cumulus cells markedly increased in patients aged 40 or over, while the fertilization rate was greatly decreased in those age group. 2. Apoptosis in cumulus cells was found in both the fertilized oocytes and unfertilized oocytes, but the incidence of apoptosis was higher in unfertilized oocytes. 3. There is no clear correlation between apoptosis in cumulus cells and the number of oocytes retrieved. However, the incidence of apoptosis was increased when the number of oocytes retrieved was 5 and fewer in comparison with $6{\sim}10$. 4. Embryo grade was significantly affected by the incidence of apoptosis in cumulus cells. 5. Pregnancy rate of IVF-ET per cycle was 29.4%, and the pregnant group had the higher fertilization rate and a significantly lower incidence of apoptosis in cumulus cells compared with the nonpregnant group. 6. When cumulus cells were used as helper cells in the co-culture of the embryo, in vitro activity of cumulus cells based on morphological change and proliferation did not influence the quality of embryo, but was closely associated with the implantation rate and pregnancy rate, which was enhanced when morphological changes and proliferation of cumulus cells was more active. 7. This difference in the outcome of IVF-ET according to in vitro activity of cumulus cells used for co-cultue was not associated with the incidence of apoptosis in cumulus cells; but rather had likely relations with the different secretion pattern of protein, which may be an embryo trophic factor by cumulus cells. Conclusion: These results suggest that the incidence of apoptosis in cumulus cells can be used in predicting oocyte qualities and the outcomes of IVF-ET. And the effect of co-culture largely depends on the in vitro activity of cumulus cells as well.