본 연구는 개 난소의 수송온도가 난자의 생존률에 미치는 영향과 체외성숙 배양시 성숙률에 미치는 영향에 대하여 검토하였다. 1. 개 난소를 4와 38에 저장 후 5시간 내로 연구실로 운반한 난소에서 채취한 난자를 체외배양 하여 생존율을 조사한 결과 체외배양 24시간째 13.2%(15/114), 77.8%(105/135)의 생존율을 보였으며, 48시간째는 0% (0/129), 72.9% (129/177) 의 생존율을 나타내어 38에 수송한 난소에서 채취한

본 실험은 원시난포의 체외배양 체계를 확립할 수 있는 가능성을 검토하기 위해 0, 5, 10, 15 및 20일령 마우스에서 분리한 난소를 20, 15, 10, 5 및 0일동안 organ culture 하여 난소의 성장률, 난자의 회수율과 성장률 및 난자의 핵성숙 단계를 조사하여, 비교 ·검토하였다. 그 결과를 요약하면 다음과 같다. 1. Organ culture 전과 후의 각 일령에 따른 난소의 면적 차이는 0일령 35.9%, 5일령 8.7%, 10 일

As an preliminary experiment for making transgenic animals producing human follicle stimulating hormone (hFSH), we tried to express recombinant hFSH gene in vitro. hFSH is a heterodimeric glycoprotein hormone produced in the anterior pituitary gland. The hormone is essential in the regulation of reproductive processes, such as follicular development and ovulation. Genes encoding the common gonadotrophin alpha subunit and FSH-specific beta subunit were inserted into retroviral vectors under the control of the rat beta actin promoter. Gene transfer to the Chinese hamster ovary (CHO) cells was done by infection of the retroviruses harvested from PT67 packaging cells transfected with recombinant retrovirus vector DNA. After selection with G4l8, PCR and RT-PCR analyses of the G4l8-resistant CHO cells showed successful transfer and expression of both and fragments of the FSH gene.

The present study was carried out for the comparative study on the collection of bovine follicular oocytes by ultrasound-guided ovum pick-up(OPU) and slaughterhouse-derived (SHD) ovary aspiration and in vitro production of bovine embryos with the follicular oncytes in Korean native cows. Bovine follicular nocytes were observed with a 6.5 MHz convex-array ultrasound transducer designed for intravaginal use and the oocytes were collected with the aspiration equipment attached to the ultrasonograph. Bovine ovaries were collected and transported in phosphate buffered saline from the local slaughterhouse, the follicular oocytes were collected by the aspiration method. The collected follicular oocytes in good quality were matured, fertilized and cultured in the media. The total number of the visible follicles and the recovery rate of follicular oocytes were increased in ultrasonography following follicle stimulating hormone(FSH) treatment in Korean native cows. The mean recovery rate of oocytes was 66.2, 52.8 and 41.7% in the FSH-OPU, non-treatment-OPU and SHD ovaries, respectively. The mean number of recorved oocytes per cow were not significantly(P<0.05) different between the FSH-OPU(14.011.54) and SHD(17.1i6.21) groups, but the numbers in both groups were significantly(P<0.05) higher than the number in the non-treatment-OPU(3.71.57) group. The mean number of usable nocytes in Grade T /11 per ovary was 6.3, 4.8 and 1.3 in the cows of the SHD, FSH-OPU and non-treatment-OPU groups, respectively. The in vitro developmental rate to the blastocyst was not significantly different between the oocytes obtained via OPU(37.1%) and SHD(29.3%). Therefore, the ultrasound-guided OPU technique can be applied to the production of excellent embryos from the high-quality cows, and for the large scale production of in vitro bovine oocytes and embryos, the SHD ovary aspiration method is valuable.

야외에서 흔히 흰불나방의 피해를 입는 7개의 기주식물에서 채집된 흰불나방의 기주식물별 용의 크기와 포란수 및 그들간의 상관을 실내사육하여 보고된 결과와 비교검토하기 위하여 실험하였다. 그 실험결과는 다음과 같다. (1) 용의 체중은 기주식물의 종류와 자웅 성에 따라 차이가 있었다. 자웅에 있어서 평균용중의 순위는 이태리푸라, 가래나무, 버즘나무, 네군도 단풍나무, 낙우송 수양버들 및 벗나무이었고 웅용에 있어서 평균용중의 순위는 낙우송, 버즘나무, 가래나무, 네군도단풍나무, 이태리 포푸라, 수양버들 및 벗나무이었다. (2) 포란수는 기생식물의 종류에 따라 차이가 있었다. 그의 평균치의 순위는 이태리포푸라, 가래나무, 낙우송, 네군도단풍나무, 버즘나무, 수양버들 및 벗나무 이었다. (3) 기주식물별 흰불나방의 용중과 포란수는 실내사육에 의하여 보고된 결과에 비하여 야외집단의 경우가 현저히 높은 수치를 나타내었다. (4) 공시기주식물은 모다 용의 크기와 포란수간에 고도의 유의한 상관관계를 나타내었다.

Fas ligand (FasL) and its receptor Fas have been implicated in granulosa cell apoptosis during follicular atresia. Although interferon-gamma (IFN-γ) is believed to be involved in the regulation Fas expression in differentiated granulosa or granulosa-luteal cells, the expression of this cytokine and its role in the regulation of the granulosa cell Fas/FasL system and apoptosis during follicular maturation have not been thoroughly investigated. In the present study, we have examined the presence of IFN-γ in ovarian follicles at different stage of development by immunohistochemistry and related their relative intensities with follicular expression of Fas and FasL, and with differences in granulosa cell sensitivity to Fas activation by exogenous agonistic Anti-Fas monoclonal antibody (Fas mAb). Although IFN-γ immunostaining was detectable in oocyte and granulosa cells in antral follicles, most intense immunoreactivity for the cytokine was observed in these cells of preantral follicles. Intense immunoreactivity for IFN-γ was most evident in granulosa cells of atretic early antral follicles where increased Fas and FasL expression and apoptosis were also observed. Whereas low concentrations of IFN-γ (10-100 U/mL) significantly increased Fas expression in undifferentiated granulosa cells (from preantral or very early antral follicles) in vitro, very higher concentrations (≥ 1,000 U/mL) were required to up-regulate of Fas in differentiated cells isolated from eCGprimed (antral) follicles. Addition of agonistic Fas mAb to cultures of granulosa cells at the two stages of differentiation and pretreated with IFN-γ (100 U/mL) elicited morphological and biochemical apoptotic features which were more prominent in cells not previously exposed to the gonadotropin in vivo. These findings suggested that IFN-γ is an important physiologic intra-ovarian regulator of follicular atresia and plays a pivotal role in regulation of expression of Fas receptor and subsequent apoptotic response in undifferentiated (or poorly differentiated) granulosa cells at an early (penultimate) stage of follicular development.

In Brassica as matter of seedling manner, they have the bilocular ovary and 20~28 seeds per silique after fertilization. Rarely some of B. juncea and yellow sarson (Brassica rapa ssp, tricolaris) have multilocular ovary. In this stdudy, the LP8 (YS-033, CGN06835) is shown tetralocular ovary as well as high seed yields. As microscope study for the different size of immature bud sections and we have known the floral meristem with already four locules in immature buds less size than 1mm of LP8. To identify of determining of tetralocular ovary formation, RNA-seq was carried out on the isolated RNA from less than 1mm and from 1mm of bud size respectively. By contrast tetralocular ovay and bilocular ovary, Chiifu is used. A total of 994 differentially expressed genes(DEGs) are detected in only LP8. Among the DEGs, we identify 18 DEGs in only immature buds of less size than 1mm. The expression patterns of 18 DEGs are validated by real time quantitative PCR and these genes are cloned and the sequence analyzed. At present, 12 candidated gene are analyzed by sequencing and there are detected by large fragment insertion as well as SNPs in sequence comparison to Chiifu. We will perform the genetic transformation of these DEG genes in Arabidopsis for relation between genes and tetralocular ovary. Our results will be helpful in understanding for mechanisms of tetraovular ovary in Brassica rapa.

본 연구는 나리 주요 품종의 종간교잡에서 생기는 삭과의 배 유 퇴화방지와 미숙배 형성 시 발생하는 이상증상을 경감하기 위해 나리 종간교잡계통의 자방배양 했을 때 cytokinin 첨가가 신초형성에 미치는 영향을 구명하고자 하였다. FO계통은 BA 2.0 ㎎/L에서 신초 발생율이 18%로 가장 효과적이었고, BA 3.0 ㎎/L와 5.0 ㎎/L에서는 12%로 다소 저조한 결과였다. TDZ 첨가 의 경우는 대조구와 같거나 비슷한 경향이었다. FA계통은 TDZ 첨가 시 대체적으로 신초형성율이 높았고 0.3 ㎎/L 처리가 효과 적이었다. 반면 BA를 첨가한 배지에서는 14.3%에서 15.7%로 대 조구와 큰 차이가 없었다. FLA계통에서 신초발생율은 생장조 절제에 의해 증가되었으나 대조구 20%와 비교했을 때 BA 20~ 24%, TDZ 22~24%로 처리효과는 미미하였다.

Acute pulmonary embolism (PE) is a common and often catastrophic disease. Early diagnosis and treatment of PE is im-portant, however, prompt diagnosis is difficult due to nonspecific clinical manifestations. On the other hand, syncope may develop as a presenting symptom of PE, and defecation activity could trigger PE. In Korea, PE associated with defecation has not been previously reported. We report on a case of acute massive PE presenting as defecation syncope in a 26-year-old female with a serous borderline tumor of the ovary.

최근 시상하부에서 생성되는 nesfatin-1/NUCB2가 섭식과 에너지 대사를 조절한다는 사실이 새롭게 밝혀졌다. 본 연구에서는 이러한 단백질이 생쥐의 생식기관에서도 발현을 하는지, 그리고 그 수용체가 생식기관 내에 존재하는 지를 확인함으로써 nesfatin-1이 생식기능에 미칠 수 있는 가능성을 알아보고자 하였다. 암컷 생쥐에서 난소와 자궁을 획득하여 conventional PCR 방법으로 NUCB2 mRNA 발현을 조사하였고, real-time PCR 방법으로 상대적인 NUCB2 mRNA 발현량을 비교 분석하였다. 난소 내 nesfatin-1 단백질의 발현 위치를 조사하기 위하여 nesfatin-1 항체를 이용한 면역조직화학염색법을 수행하였으며, biotin conjugated nesfatin-1을 이용하여 nesfatin-1 결합 부위를 확인하였다. 또한 생식소 내 NUCB2 mRNA 발현이 성선자극호르몬에 의해 영향을 받는지 알아보기 위해 PMSG 투여 후 NUCB2 mRNA 발현량을 조사하였다. 실험 결과, 생쥐의 난소와 자궁에서 확인된 NUCB2 유전자가 시상하부에서 만큼이나 많은 양이 발현되고 있었다. 면역조직화학적 염색 결과, nesfatin-1 단백질은 협막세포와 대부분의 기질세포에서 발현되었고, 일부 황체세포에서도 발현이 확인되었다. 반면, 난포 내 과립세포에서는 발현되지 않았으나, 특정 난포 내 난자에서는 발현됨을 확인하였다. 한편, nesfatin-1 단백질의 결합 부위는 난소 백막 주위의 기질세포와 협막세포에서 관찰되었다. 또한 PMSG 투여 후 난소와 자궁에서 NUCB2 mRNA의 발현이 유의하게 증가함을 확인하였다. 이상의 결과에서 난소 내 nesfatin-1 단백질의 발현과 그 결합 부위의 존재는 nesfatin-1이 뇌에서 뿐만 아니라 생식기관에서도 국부조절인자로써 중요한 역할을 할 것으로 사료되며, 앞으로 생식기관에 미치는 nesfatin-1의 역할을규명하기위한더많은연구가필요하다고판단된다.

Insulin like3 (INSL3, Relaxin like factor) is a critical regulator in testis translocation through Leucin rich G-protein coupled receptor 8 (Rxfp2,LGR8) during embryogenesis. In female, INSL3 and their receptor expressed in growing follicle and revealed their function in oogenesis. However, its role is not much evaluated. 6 weeks old C57BL/6 female mice used for measure the expression of INSL3 mRNA and their receptor expression by follicular stage. Follicle cell specific RNA were got from the theca cell and granulose cells which were isolated using LCM. To know the role of INSL 3 in theca cell, Rxpf2 were overexpressed during primary culture of theca cells, which were isolated from tertiary follicles after 12 hr PMSG injection and transduced the Rxfp2 using lenti virus. After 10 days of culture, the proliferation of theca cells was analyzed using EdU. Using Alzet osmotic pumps INSL3 was administered for 3 days into the ovary during superovulation induction. The INSL3 mRNA levels were significantly high in the theca cells of preovulator follicles after hCG injection. But granulose cells showed decreasing expression by growth of follicle. INSL3 stimulated the proliferation of theca cells in vitro which overexpressed the Rxpf2. By the administration of INSL3 into ovary caused the dramatical decrease the number of ovulated oocyte. Based on these results, we know that INSL3 stimulates the theca cell proliferation in as follicle stage specific manners, and estrogen is a modulator of this INSL3 mRNA expression. It suggests that the disturbance of the expression regulation of INSL3 and its receptor cause the unregulated theca cell proliferation and failing the rupture of grown follicle.

일부 식물성 에스트로겐(phytoestrogen)의 경우, 긍정적인 효과를 갖는 것으로 보이지만, 대부분의 내분비계 장애 물질(endocrine disruptor 또는 endocrine disrupting compound, EDC)은 노출된 개체의 내분비계를 교란시켜 인간이나 야생 동물의 건강에 해로운 것으로 알려져 있다. 선행 연구에서 본 연구자들은 사춘기 전에 단기간으로 식물성 에스트로겐인 genistein(GS)을 투여했을 때 암컷 흰쥐의 생식계가

본 논문은 포유동물 난소에서의 신속한 프로게스테론(P4) 선호 전달경로에 관해 현재 통용되는 지식을 요약하였다. P4는 안드로겐과 에스트로겐 합성 과정에서의 중요한 중간 산물이면서 그 자체로도 배란, 난포폐쇄(atresia), 황체형성과정(luteinization)에서 결정적인 역할을 하며, 모든 포유동물의 초기 임신 유지에 필수적이다. 이와 같은 생리적인 중요성에도 불구하고 포유동물 난소에서의 정확한 P4 작용기작은 아직까지도 완전히 알려져 있지 않다