The study was carried out to investigate the effects of morphology, reproductive cycle, incubation time and activation of oocytes on in vitro maturation of cat oocytes and development of IVM/IVF embryos. The results were summarized as follows: 1. When recovered from ovaries collected at different stages of the reproductive cycle (inactive, follicular and luteal stage), the developmental rates of oocytes to GV and MI stage were 72.5% and 27.5%, 57.5% and 7.5%, 62.5% and 17.5%, respectively. 2. The developmental rates of oocytes with cumulus cells to GV and MI stage in different conditions of incubation (5% CO₂, 95% O₂ and 10% CO₂, 90% O₂) were 70.0% and 27.5%, 52.5% and 20.0%, 55.0% and 12.5%, respectively. 3. The developmental rates to GV and MI oocytes when cultured at different time of incubation (17∼20, 21∼24, 25∼28 and 29∼32 h) were 67.5% and 20.0%, 67.5% and 30.0%, 62.5% and 22.5%, 65.0% and 15.0%, respectively. 4. The fertilization and cleavage rates of freshly collected oocytes with and without cumulus cells were 72.5% and 25.0%, 37.5% and 7.5%, respectively. The rates were greater in oocytes with cumulus cells than those without cumulus cells. 5. The fertilization and cleavage rates of oocytes recovered from ovaries collected at different stages of the reproductive cycle (inactive, follicular and luteal stage) were 75.0% and 25.0%, 40.0% and 7.5%, 50.0% and 15.0%, respectively.

본 연구는 소형 고양이의 불임 해결과 체외수정란을 생산하기 위한 방안으로서 난자의 형태, 번식주기, 배양시간 및 활성화 처리가 난포란의 체외수정 및 체외발생에 미치는 영향을 조사하였다. 1. 신선 및 salt에 보존한 난소로부터 회수한 난구세포부착 및 나화 난자를 각각 배양했을때 체외수정율 및 분할율은 65.7%와 17.1%, 28.6%와 8.6% 및 57.1%와 13.3%, 23.3%와 3.3%로서 난구세포 부착 신선난자가 나화 난자에 비해 높은 체외

도심 지역인 서울시청 부근과 공단지역인 안산 반월 공단지역 내에 서식하는 비둘기의 알,새끼, 성조를 대상으로 각 발달 단계에 따른 납과 카드뮴의 체내 축적 농도를 비교하고 각 지역의 오염 수준을 파악하고자 수행하였다 조사 결과, 납 및 카드뮴 농도는 뼈, 신장, 간, 허파 등 모든 조직에서 새끼보다 어미가 높았으며,특히 납은 뼈에서, 카드뮴은 신장에서 현저하게 높았다. 서울의 경우, 납 농도는 알보다 새끼의 뼈 조직에서 약 3배 높았고, 새끼보다 성조의

This study was performed to find out the optimal larval stage among trochophore, D-shaped and umbo stage larvae and the desirable protective additive such as fructose, glucose, sucrose and trehalose with cryoprotectant for cryopreservation of surf clam, Spisula sachalinensis larvae. Dimethyl sulfoxide and ethylene glycol were used as cryoprotectant and each cryoprotectant was made to 2.0 M with previous protective additives. The larvae were immersed in the preparations waited for 15 minutes to reach equilibration, and then frozen in a program freezer (-35) and liquid nitrogen (-196). The freezing rate of 1.0 /min. was used for cryopreservation of trochophores before seeding temperature (-12). The survival rate of frozen-thawed larvae increased as larval developing and that of umbo stage larvae was the highest as 96.1 1.0%. The presence of lower concentration of disaccharides as sucrose or trehalose significantly enhanced survival rate when mixed with cryoprotectants (P<0.05). The results of our study indicate that desirable developmental stages of larvae and protective additive for cryopreservation are the umbo stage larvae and 0.2 M sucrose, respectively.

산림발달에 따른 조릿대 임분의 구조적 특성을 구명하기 위해 전남 광양의 백운산 지역내 천연임분과 1987년 벌채임분, 1993년 벌채임분에서의 조릿대 발생임분과 비발생 임분에 대해 식생조사를 실시하였다. 또한, 상층 수목의 생장에 대한 조릿대의 영향을 조사하기 위해 조릿대가 분포하는 임분과 분포하지 않는 임분에서 공통적으로 중요도가 높은 졸참나무와 서어나무에 대한 연륜 생장량 분석을 실시하였다. 조사연구지역의 식생형은 조릿대 우점 식생형과 조릿대 소점 또는 무점 식생형으로 크게 분류되었으며. 전자의 경우 졸참나무-서어나무/조릿대 군락, 때죽나무-쪽동백나무/조릿대 군락, 층층나무-산뽕나무/조릿대 군락, 느티나무-조릿대 군락 등 4개의 군락으로 분류되었고, 후자의 경우는 졸참나무/비목군락, 층층나무/비목 군락, 졸참나무/당단풍/물참대 군락, 물푸레나무-졸참나무/비목/회잎나무 군락, 산딸기-산수국 군락, 산딸기-싸리 군락 등 6개 군락으로 분류되었다. 임분발달 단계에 따라서는 천연임분의 경우 졸참나무와 서어나무 중심의 낙엽활엽교목과 하층식생으로서 조릿대가 우점하는 반면, 벌채임분의 경우 하층식생으로서의 조릿대 출현밀도가 다양하게 나타났다. 졸참나무와 서어나무의 초기생장기에는 조릿대가 발생하지 않은 임분에서 자라는 개체들의 연륜폭이 조릿대 발생임분에서 자라는 개체들의 연륜폭보다 넓었으나 약 30년을 전후하여 반대 현상이 나타나고 있어 조릿대에 의한 생장감소는 없는 것으로 판단된다.

This experiment was carried out to investigate the ovarian responses of the ovulation point, ovarian weight and size, the number of ovarian follicles and collected embryos, and to study the effects of the developmental stages (oocytes, 2-4 cell. 8-16 cell and morulae), additional levels of Ficoll (0, 15, 30%) on the survival rate (FDA-test) of rat embryos frozen in vitrification solution (20% glycerol + 10% ethylene glycol + 10% sucrose). Sunanarized results was as follows; 1. The mean ovulation point per head was 7, and the weight of ovaries was 0.03g. The size of ovary was 5.9 mm(L) and 4.6 mm(W), and the number of ovarian follicles over and below 2 mm was 4.7 and 8.7, respectively. The number of the collected embryos per head was 5.5 (79%). 2. 2. The FDA score of embryos frozen in 20 G 10 E 10 S without Ficoll was 2.8 (oocyte), 2.6 (2-4 cell), 3.9 (8-16 cell) and 3.6 (morula), respectively. However, there were no significant differences among treatments. 3. The FDA score of embryos frozen in 20 G 10 E 10 S with 15% Ficoll was 3.4 (oocyte), 4.0 (2-4 cell), 4.7 (8-16 cell) and 4.8 (morulae), respectively (P>0.05). 4. The FDA score of embryos frozen in 20 G 10 E 10 S with 30 % Ficoll was 3.7 (oocyte), 3.2 (2-4 cell), 4.4 (8-16 cell) and 4.4 (morulae), respectively (P>0.05). 5. As shown in the above results, the higher survival rate was obtained in the treatment of 15% Ficoll than that of 30%. And the survival rate (FDA-test)of the oocytes and 2-4 cell stages of the rat embryos was lower than that of 8~16 cell and morulae stages. It was considered that 8-16 cell and morulae could be available for the successful freezing by vitrification of rat embryos with 15% Ficoll except for oocytes.

The present experirnents on cryopreservation were carried out to investigate effect of solution toxicity, equilibration time and cell stages on the post-thaw survival of mouse morulae and blastocyst embryos cryopreserved by vitrification in EFS solution. The mouse embryos were exposed to the EFS solution in one step at room temperature, kept in the EFS solution during different period for toxicity test, vitrified in liquid nitrogen and thawed rapidly. After the mouse morulae embryos were exposed to EFS solution for 2 and 5 ruin. at room temperature and then they were washed in 0.5 M sucrose solution and basal mediurn(D-PBS + 10% FCS), they were cultured to examined cryoprotectant toxicity induced injury during exposure, most of embryos developed to expanded blastocysts(100 and 90.0%). However, when the exposure time was extended to 10 and 20 min, these development rates dropped dramatically in 10 ruin. (75.0%) and 20 ruin. (4.5%), respectively. When the compacted morulae were vitrified in EFS solution after equilibration for 2 and 5 min, the embryos have developed to normal blastocyst following thawing, washing and culture processes was 89.3 and 89.6%. However, when the exposure time was expanded to 10 ruin, this survival rate dropped to 68.8%. When the blastocyst were vitrified in EFS solution after equilibration for 2, 5 and 10 minutes, the survival rate of embryos which developed to normal blastocyst following thawing and culture processing were 58.5, 46.7 and 22.4%, respectively. The optimal time of equilibration of mouse morula and blastocysts in EFS solution seemed o be 2 and 5 ruin.

The post-thaw survival of mouse embryos of the various developmental stages was determined after cryopreservation by vitrification in a solution containing ethylene glycol, Ficoll and sucrose (EFS). All the embryos were equilibrated for 2 minutes just prior to freezing. The number of blastomeres during in vitro development was counted by nuclei higher rates of post-thaw survival were obtained from the embryos of 2-cell(92.2%), 8-cell(77.2%) or morula stage(90.0%) than those of blastocyst stage(62.7%). The number of blastomeres per embryo following in vitro culture for 24 hours was significantly(P<0.05) smaller as 66.0f22.3 in vitrified and thawed morulae than fresh morulae(91.712.2).

키틴생합성저해제인 diflubenzuron을 톱다리개미허리노린재(Riptortus clavatus Thunberg)에 미량국소처리하였을 때 충태 및 영기에 따른 약제의 감수성차이와, 종령약충 처리후 우화율 및 우화성충에 미치는 영향을 조사하였다. Diflubenzuron은 살람효과를 보였으며 산란후 12시간내의 알은 산란후 48~60시간의 알보다 감수성이 높았으나, 알의 나이에 관계없이 처리된 알의 배는 정상적으로 발육하였다. 영기별 감수성은 영기가 진행될수록 낮아져 1령약충이 2령에 비하여 1.5배, 3령약충에 비하여 18.2배, 4령약충에 비하여 39.4배, 5령약충에 비하여 42.4배 높은 감수성을 나타내었다. 종령약충에 처리하였을 때 우화율과 우화성충의 체중, 수명 및 산란율은 현저히 감소하였다.

Background : Agastache rugosa (A. rugosa), belonging to the Lamiaceae family, is a medicinal plant mainly distributed in Korea and contains various phenolic compounds revealing anti-fungal and anti-HIV properties. This study is aim to investigate change in phenylpropanoid content of flowers at different developmental stages using high performance liquid chromatography (HPLC) and quantitative real time polymerase chain reaction (qRT-PCR). Methods and Results : The variation in the transcriptional level of phenylpropanoid biosynthetic genes and phenylpropanoid contents in the flowers of A. rugosa at different developmental stages was analyzed. The transcript levels of phenylpropanoid biosynthesis genes, including ArPAL (phenylalanine ammonia-lyase), ArC4H (cinnamate 4-hydroxylase), and ArCHS (Chalcone synthase), were high in flowers at 1st stage compared with flowers at 2nd and 3rd stages. On the other hand, the expression levels of flavonoid biosynthesis genes, including ArTAT (tyrosine amino transferase), ArHPPR (hydroxyl phenylpyruvate reductase), and ArRAS (rosmarinic acid synthase), were higher in flowers at 3rd stage than those of flowers at 1st and 2nd. These results were consistent with HPLC analysis revealing that most phenolic compounds were higher in flowers at 1st and 2nd stage but the level of rosmarinic acid was the highest in 3rd stage. Conclusion : Our findings provide the information on change in phenylpropanoid biosynthesis in A. rugosa flowers at different developmental stages.

Cytological changes of the epithelial cells according to the developmenatal phases of the seminal vesicle related to the spermatogenic stages in the testicular lobules during spermagenesis in male Neptunea (Barbitonia) cumingii (Gastropoda: Buccinidae) were investigated monthly by electron microscopical and histological observations. N. (B) cumingii is dioecious, and an internal fertilization species. The male genital organ is located near the tentacles. The spermatozoon is approximatley 50 μm in length. The axoneme of the tail flagellum consists of nine pairs of microtubles at the periphery and one pair at the center. The process of germ cell development during spermatogenesis can be divided into five succesive stages: (1) spermatogonia, (2) primary spermatocytes, (3) secondary spermatocytes, (4) spermatids, and (5) spermatozoa. A considerable amount of spermatozoa make their appearance in the testicular lobules (or acini) and some of them are tranported from the testis towards the seminal vesicles until late July. In this study, the developmental phases of the epithelial cells of the seminal vesicles of N. (B.) cumingii could be classified into four phases: (1) S-I phase (resting), (2) S-Ⅱphase (early accumulating), (3) S-Ⅲ phase (accumulating), and (4) S-IV phase (spent). However, in case of N. (B.) arthritica cumingii, the developmental phases of the seminal vesicle were devided into three phases: (1) resting, (2) accumulating and (3) spent. Granular bodies in the inner layer of the seminal vesicles are involved in resorption of digestion of residual spermatozoa.

Purine metabolism is known factor for nuclear maturation of oocytes through both follicle cells and oocyte itself. However, it is largely unknown the roles of purine metabolism in the oocyte competence for fertilization and early development. In this study, the effects of adenosine in oocyte competence for development were examined using adenosine and its synthetic inhibitors. Adenosine treatment from GV intact stage for 18 hr (fGV) caused of decrease the fertilization rate but of increase the cleavage rate compared from the other stage treatment groups. Hadacidin did not effect on fertilization rate but increased cleavage rate without stage specificity. Adenosine did not block the effects of hadacidin with the exception of fGV group. By the inhibition of purine synthetic pathways the fertilization rate was decreased in the fGV and fGVB groups but increased in the fMII group. Exogenous adenosine caused of decrease fertilization rate in the fGVB group but increase in the fMII group. Cleavage rate was dramatically increased in the adenosine treatment with synthetic inhibitors. It means that the metabolism of purine has stage specific effects on fertilization and cleavage. Exogenous adenosine had only can improve oocyte developmental competence when it treated at GV intact stage. On the other hand, endogenous synthesis in all maturation stage caused of increase the cleavage rate without effects on fertilization. These data suggest that adenosine at GV stage as a paracrine fashion and inhibitions of endogenous adenosine in all stage improve oocyte developmental competence..

Fish larvae are immediately exposed to microbes from hatching to maturation of their lymphoid organs, therefore effective innate mechanisms is very important for survival in such an environment. The key component of innate immune system, C3 is central protein of all activation pathways of the complement system, leading to inflammatory reactions, such as opsonisation, chemotaxis, and cell lysis of pathogens. Although, innate mechanisms is essential for survival in the early stage of development, little is known about defence mechanisms. In this study, the alignment analysis showed that amino acid sequence of C3 from olive flounder liver EST homologous to other known C3 sequences with 73-99% identity. Also, we examined the tissue distribution of olive flounder C3 and analyzed expression pattern from the fertilized egg until 28 days post hatching. As a result, olive flounder C3 mRNA was expressed only in the liver and the mRNA level more increased as developmental proceed during the early stage. These results may suggest that olive flounder C3 plays an important function in the early immune response of olive flounder larvae.

The seminiferous epithelium cycle and developmental stages of spermatids in Clethrionomys rufocanus were observed under a light microscope. The seminiferous epithelium cycle was divided into 8 stages. Type Ad spermatogonia appeared through all stages. Type Ap, In, and B spermatogonia appeared in stages Ⅰ, Ⅱ, Ⅲ, and Ⅳ. In the first meiosis prophase, the leptotene spermatocytes appeared from stage Ⅴ, the zygotene spermatocytes in stages Ⅰ, Ⅵ, Ⅶ, Ⅷ, the pachytene spermatocytes from stages Ⅱ to Ⅵ, the diplotene spermatocytes in stage Ⅶ. The meiotic figures and interkinesis spermatocytes were observed in stage Ⅷ. Developing spermatids were subdivided into 10 steps, based on the morphological characteristics such as the acrosome formation changes in spermatozoa, nucleus, cytoplasm, and spermiation changes. The C. rufocanus spermatocytogenesis and spermiogenesis results displayed similar results with Apodemus agrarius coreae and A. speciosus peninsulae. Considering all the results, the spermatogenesis may be useful information to analyze the differentiation of spermatogenic cells and the breeding season.