• Title/Summary/Keyword: developmental rate

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Cryopreservation of Bovine IVM/IVF/IVC Hatched Blastocysts (체외생산된 소 완전탈출 배반포기배의 동결보존)

  • Lee, K.S.;Kim, E.Y.;Yi, B.K.;Nam, H.K.;Yoon, S.H.;Park, S.P.;Lim, J.H.
    • Korean Journal of Animal Reproduction
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    • v.23 no.2
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    • pp.141-148
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    • 1999
  • This study was to test whether the viability of bovine hatched blastocysts (HBs) can be maintained after vitrification and thawing. The HBs were produced in vitro at Day 9 and Day 10 after IVF, and they were classified to small (S-HBs; ø$\leq$300 ${\mu}{\textrm}{m}$) and large(L-HBs; ø>300 ${\mu}{\textrm}{m}$) on the basis of embryo diameter using eyepiece micrometer. As freezing solution, we used EFS35 which consisted of 35% ethylene glycol (EG), 18% ficoll, 0.3 M sucrose and 10% FBS added in mDPBS. Vitrification was taken by two-step method, the HBs were equilibrated in 10% EG for 5 minutes and then shortly exposed in EFS35 and plunged into L$N_2$for 30~45 sec. After thawing, the survival rates were assessed by the re-expansion of the blastocoel during 2 h and 16 h of culture. The results obtained in these experiments were summarized as follows; 1) When the blastocysts(40.8%) recovered at Day 8 after IVF were further cultured for 24 h(Day 9 after IVF) and 48 h(Day 10 after IVF), the rates of HBs were 20.5% and 6.7%, respectively. Also, the total cell number of HBs on Day 9 was significantly higher than that of HBs on Day 10 (p<0.01). 2) When the effects of freezing solution to the survival of Day 9 L-HBs were examined, the rate of vitrified group (75.7%) was significantly lower than 100% of control and exposed group(p<0.05). 3) When the survival rates of vitrified HBs according to size and developmental age were examined, the data of L-HBs (75.5%) and S-HBs(63.6%) on Day 9 were slightly higher than those of L-HBs(64.3%) and S-HBs(60.7%) on Day 10. 4) Also, when the in vitro survival of Day 9 HBs was evaluated under different culture condition after thawing, the result in culture medium only (79.3%) was significantly higher than 43.2% in co-culture group (p<0.05). These results demonstrated that bovine HBs can be successfully cryopreserved by two-step vitrification method using EFS35.

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The embryological studies on the interspecific hybrid of ginseng plant (Panax ginseng x P. Quiuquefolium) with special references to the seed abortion (인삼의 종간잡종 Panax ginseng x P Quinquefoilium의 발생학적 연구 특히 결실불능의 원인에 관하여)

  • Jong-Kyu Hwang
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.5 no.1
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    • pp.69-86
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    • 1969
  • On the growing of the interspecific hybrid ginseng plant, the phenomena of hybrid vigoures are observed in the root, stem, and leaf, but it can not produce seeds favorably since the ovary is abortive in most cases in interspecific hybrid plants. The present investigation was undertaken in an attempt to elucidate the embryological dses of the seed failure in the interspecific hybrid of ginseng (Panax Ginseng ${\times}$ P. Quinque folium). And the results obtained may be summarized as follows. 1). The vegetative growth of the interspecific hybrid ginseng plant is normal or rather vigorous, but the generative growth is extremely obstructed. 2). Even though the generative growth is interrupted the normal development of ovary tissue of flower can be shown until the stage prior to meiosis. 3). The division of the male gameto-genetic cell and the female gameto-genetic cell are exceedingly irregular and some of them are constricted prior to meiosis. 4). At meiosis in the microspore mother cell of the interspecific hybrid, abnormal division is observed in that the univalent chromosome and chromosome bridge occure. And in most cases, metaphasic configuration is principally presented as 23 II+2I, though rarely 22II+4I is also found. 5). Through the process of microspore and pollen formation of F1, the various developmental phases occur even in an anther loclus. 6). Macro, micro and empty pollen grains occur and the functional pollen is very rare. 7). After the megaspore mother cell stage, the rate of ovule development is, on the whole, delayed but the ovary wall enlargement is nearly normal. 8). Degenerating phenomena of ovules occur from the megaspore mother cell stage to 8-nucleate embryo sac stage, and their beginning time of constricting shape is variously different. 9). The megaspore arrangement in the parent is principally of the linear type, though rarely the intermediate type is also observed, whereas various types, viz, linear, intermediate, Tshape, and I shape can be observed in hybrid. 10). After meiosis, three or five megaspore are some times counted. 11). Charazal end megaspore is generally functional in the parents, whereas, in F1, very rarely one of the center megaspores (the second of the third megaspore) grows as an embryo sac mother cell. 12). In accordance with the extent of irregularity or abnormality in meiosis, division of embryo sac nuclei and embryo sac formation cause more nucellus tissue to remain within th, embryo sac. 13). Even if one reached the stage of embryo sac formation, the embryo sac nuclei are always precarious and they can not be disposed to theil proper, respective position. 14). Within the embryo sac, which is lacking the endospermcell, the 4-celled proembryo, linear arrangement, is observed. 15). Through the above respects, the cause of sterile or seed failure of interspecific hybrid would be presumably as follows, By interspecific crossing gene reassortments takes place and the gene system influences the metabolism by the interference of certain enzyme as media. In the F1 plant, the quantity and quality of chemicals produced by the enzyme system and reaction system are entirely different from the case of the parents. Generally, in order to grow, form, and develop naw parts it is necessary to change the materials and energy with reasonable balance, whereas in the F1 plant the metabolic process becomes abnormal or irregular because of the breakdown of the balancing. Thus the changing of the gene-reaction system causes the alteration of the environmental condition of the gameto-genetic cells in the anther and ovule; the produced chemicals cause changes of oxidatio-reduction potential, PH value, protein denaturation and the polarity, etc. Then, the abnormal tissue growing in the ovule and emdryo sac, inhibition of normal development and storage of some chemicals, especially inhibitor, finally lead to sterility or seed failure. Inconclusion, we may presume that the first cause of sterile or seed abortion in interspecific hybrids is the gene reassortment, and the second is the irregularity of the metabolic system, storage of chemicals, especially inhibitor, the growth of abnormal tissue and the change of the polarity etc, and they finally lead to sexual defect, sterility and seed failure.

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Effect of Early Pregnant Heifer as Donor on the Ovum Pick-Up Derived Oocyte Aspiration and Embryo Production (초기 임신우의 공란우 활용이 초음파 유도 난자 채취 및 수정란 생산에 미치는 영향)

  • Jin, Jong-In;Kwon, Tae-Hyeon;Choi, Byeong-Hyun;Kim, Sung-Soo;Jo, Hyun-Tea;Bang, Jae-Il;Kim, Sam-Chul;Cho, Kyu-Woan;Lee, Jung-Gyu;Kong, Il-Keun
    • Journal of Embryo Transfer
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    • v.26 no.1
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    • pp.19-25
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    • 2011
  • This study was carried out to evaluate the effect of early pregnant cow as donor for Ovum Pick-Up (OPU) derived oocyte aspiration and embryo production in Holstein heifers. Four non-pregnant and 2 pregnant Holstein heifers were used as donor and then carried out total 17 OPU session for 10 weeks (2 times per week). Recovered cumulus-oocyte-complexes (COCs) were classified into 4 grade by oocyte cytoplasm and cumulus cells and matured in vitro in TCM-199 supplemented with 10% FBS, 10 mg/ml FSH and 1 mg/ml estradiol in 5% $CO_2$ and over 99% humidity for 24 h. After 24 h co-incubation with post-thaw sperm, the presumed zygotes were cultured in CR1aa medium with 4 mg/ml BSA for 3 days and then changed CR1aa medium with 10% of FBS for another 3~4 days. The Mean number of aspirated follicles and collected oocytes in the early stage pregnant and non-pregnant heifers were $13.0{\pm}4.3$ and $10.6{\pm}3.9$, $5.4{\pm}3.4$ and $7.7{\pm}3.6$ per session, respectively. Rate of collected oocyte from aspirated follicles were 59.2% and 50.5%, respectively. The average number of good quality oocytes (Grade I and II) in the early stage pregnant and non-pregnant heifers was $3.7{\pm}2.7$ and $4.9{\pm}2.6$ (Mean${\pm}$SD). Cleavage and blastocyst developmental rates in Grade I and II were 22.2% and 25.5%, and then $1.7{\pm}0.9$ and $1.4{\pm}1.1$ blastocyst per session, respectively. In conclusion, OPU technology can be used in early stage pregnant and non-pregnant heifers without any problem and so applied OPU derived embryo production to maximize the ability of genetically valuable females.

The Developmental Effects of Radiation on ICR Mouse Embryos in Preimplantation Stage (착상전기(着床前期)에 있어서 ICR Mouse의 태아(胎兒)에 대한 방사선(放射線) 개체(個體) Level 영향(影響)의 연구(硏究))

  • Gu, Yeun-Hwa
    • Journal of Radiation Protection and Research
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    • v.21 no.4
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    • pp.273-284
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    • 1996
  • Embryos and fetuses are more sensitive to various environmental agents than are adults or children. The biological effects such as intrauterine death and malformation are closely connected with prenatal exposure very various agents. The sensitivity of these embryonic/fetal effects depends on the stage of pregnancy. From the viewpoint of fetal development, embryonic and fetal stages can be divided into three stages : Preimplantation, organogenetic and fetal. Each stage corresponds to 0 to 4.5days, 4.5 to 13.5days, and 13.5days of gestation in mice, respectively. Many studies on the biologcal effects of mice irradiated by ${\gamma}-rays$ at various stages during organogenesis and fetal period have been performed. Based on these results, the dose-effect and dose-response relationships in malformations, intrauterine death, or retardation of the physical growth have been practically modeled by the ICRP(International Commission on Radiological Protection) and other international bodies for radiation protection. Many experimental studies on mice have made it clear that mice embryos in the preimplantation period have a higher sensitivity to radiation for lethal effects than the embryos/fetuses on other prenatal periods. However, no eratogenic effects of radiation at preimplantation stages of mice have been described in many textbooks. It has been believed that 'all or none action results' for radiation of mice during the preimplantation period were applied. The teratogenic and lethal effects during the preimplantation stage are one of the most important problems from the viewpoint of radiological protection, since the preimplantation stage is the period when the pregnancy itself is not noticed by a pregnant woman. There are many physical or chemical agents which affect embryos/fetuses in the environment. It is assumed that each agents indirectly effects a human. Then, a safety criterion on each agent is determined independently. The pregnant ICR mice on 2, 48, 72 or 96 hours post-conception (hpc), at which are preimplantation stage of embryos, were irradiated whole body Cesium-gamma radiation at doses of 0.1, 0.25, 0.5, 1.5, and 2.5 Gy with dose rate of 0.2 Gy/min. In the embryos from the fetuses from the mice irradiated at various period in preimplantation, embryonic/fetal mortalities, incidence of external gross malformation, fetal body weight and sex ratio were observed at day 18 of gestation. The sensitivity of embryonic mortalities in the mice irradiated at the stage of preimplantation were higher than those in the mice irradiated at the stage of organogenesis. And the more sensitive periods of preimplantation stage for embryonic death were 2 and 48 hpc, at which embryos were one cell and 4 to 7 cell stage, respectively. Many types of the external gross malformations such as exencephaly, cleft palate and anophthalmia were observed in the fetuses from the mice irradiated at 2, 72 and 96 hpc. However, no malformations were observed in the mice irradiated at 48 hpc, at which stage the embryos were about 6 cell stage precompacted embryos. So far, it is believed that the embryos on preimplantation stage are not susceptible to teratogens such as radiation and chemical agents. In this study, the sensitivity for external malformations in the fetuses from the mice irradiated at preimplantation were higher than those in the fetuses on stage of organogenesis.

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Effect of 6-Hydroxydopamine (6-OHDA) on the Expression of Hypothalamus-Pituitary Axis Hormone Genes in Male Rats (수컷 흰쥐의 시상하부-뇌하수체 축 호르몬 유전자 발현에 미치는 6-Hydroxydopamine(6-OHDA)의 영향)

  • Heo, Hyun-Jin;Ahn, Ryun-Sup;Lee, Sung-Ho
    • Development and Reproduction
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    • v.13 no.4
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    • pp.257-264
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    • 2009
  • A neurotoxin, 6-hydroxydopamine (6-OHDA) has been widely used to create animal model for Parkinson's disease (PD) due to its specific toxicity against dopaminergic (DA) neurons. Since DA signals modulate a broad spectrum of CNS physiology, one can expect profound alterations in neuroendocrine activities of both PD patients and 6-OHDA treated animals. Limited applications of 6-OHDA injection model, however, have been made on the studies of hypothalamuspituitary neuroendocrine circuits. The present study was performed to examine whether blockade of brain catecholamine (CA) biosynthesis with 6-OHDA can make any alteration in the transcriptional activities of hypothalamus-pituitary hormone genes in adult male rats. Three-month-old male rats (SD strain) were received 6-OHDA ($200{\mu}g$ in $10{\mu}\ell$ of saline/animal) by intracerebroventricular (icv) injection, and sacrificed after two weeks. To determine the mRNA levels of hypothalamuspituitary hormone genes, total RNAs were extracted and applied to the semi-quantitative RT-PCRs. The mRNA levels of tyrosine hydroxylase (TH), the rate-limiting enzyme for the catecholamine biosynthesis, were significantly lower than those from the control group (control:6-OHDA=1:0.72${\pm}$0.02AU, p<0.001), confirming the efficacy of 6-OHDA injection. The mRNA levels of gonadotropin-releasing hormone (GnRH) and corticotropin releasing hormone (CRH) in the hypothalami from 6-OHDA group were significantly lower than those from the control group (GnRH, control:6-OHDA=1:0.39${\pm}$0.03AU, p<0.001; CRH, control:6-OHDA=1:0.76${\pm}$0.07AU, p<0.01). There were significant decreases in the mRNA levels of common alpha subunit of glycoprotein homones (Cg$\alpha$), LH beta subunit (LH-$\beta$), and FSH beta subunit (FSH-$\beta$) in pituitaries from 6-OHDA group compared to control values (Cg$\alpha$, control:6-OHDA=1:0.81${\pm}$0.02AU, p<0.001; LH-$\beta$, control:6-OHDA=1:0.68${\pm}$0.04AU, p<0.001; FSH-$\beta$, control:6-OHDA=1:0.84${\pm}$0.05AU, p<0.001). Similarly, the level of adrenocorticotrophic hormone (ACTH) transcripts from 6-OHDA group was significantly lower than that from the control group (control: 6-OHDA=1:0.86${\pm}$0.04AU, p<0.01). The present study demonstrated that centrally injected DA neurotoxin could downregulate the transcriptional activities of the two hypothalamus-pituitary neuroendocrine circuits, i.e., GnRH-gonadotropins and CRH-ACTH systems. These results suggested that hypothalamic CA input might affect on the activities of gonad and adrenal through modulation of hypothalamus-pituitary function, providing plausible explanation for frequent occurrence of sexual dysfunction and poor stress-response in PD patients.

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Reports on bionomical characteristics of Mellicta ambigua (여름어리표범나비(Mellicta ambigua (Menetries))의 생태적 특성에 관한 보고)

  • Kim, Se-Gwon;Nam, Gyoung-Pil;Kim, Nam-Ee;Bae, Kyoung-Sin;Choi, Young-Cheol;Lee, Sang-Hyun
    • Journal of Sericultural and Entomological Science
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    • v.52 no.2
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    • pp.110-116
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    • 2014
  • Recently the number of the butterflies, Mellicta ambigua, had been decreasing rapidly, and already disappeared at many habitat. In this studies, we investigated ecological environment of Mellicta ambigua for preparing of primary research data recovering habitat, and studied on bionomical characteristics. Two different habitat, Jindo and Inje, were selected for investigation of ecological environment. We investigated four times during 3-month, from June to August in 2012. In Jindo, we observed more than 100 butterflies and a lot of host plants, Melampyrum roseum var. japonicum. But only 5 butterflies and only a few host plants, Veronicastrum sibiricum were observed in Inje. We could not observe the eggs, the larva and pupa on the host plants at all. For finding of bionomical characteritics, we reared butterflies at natural conditions. Collected 3-female butterflies from Jindo laid 465 eggs on the leaves of 3-host plants, Veronicastrum sibiricum. 120 ~ 186 eggs per each female were laid in the shape of cluster. An egg was globular shape, 0.6 mm diameter and 0.7 mm height. The egg periods were $9.96{\pm}0.4days$ after ovipositioning, and the hatchability was 95.% at natural condition. The larval periods were $4.1{\pm}0.6days$ (1st instar), $2.1{\pm}1.0days$ (2nd), $8.1{\pm}0.7days$ (3rd), $239.2{\pm}10.9days$ (4th), $12.3{\pm}1.3days$ (5th), $17.1{\pm}1.1days$ (6th), $10.5{\pm}1.0days$ (7th) each other. The larva of 4th instar overwintered in the nest that had been made into the leaf of host plant with secreted thread as a group until early March next year. In the early March next year, overwintered larva went around their nest in search of host plants, and went to other host plants, Veronica persica and Plantago asiatica, sometimes. The overwintered larva of Mellicta ambigua could grow up on two other host plants normally. In the following experiment, the butterflies of Mellicta ambigua laid eggs on the leaves of Plantago asiatica, but the 1st instar larva from eggs died all. The headwidth of each developmental larval stage were $0.28{\pm}0.02mm$ (1st), $0.45{\pm}0.02mm$ (2nd), $0.58{\pm}0.02mm$ (3rd), $0.75{\pm}0.03mm$ (4th), $0.89{\pm}0.05mm$ (5th), $1.23{\pm}0.06mm$ (6th), $2.13{\pm}0.11mm$ (7th). The pupal ratio was 92.0%. The pupal period were $9.1{\pm}1.6days$, and the emergence rate was 88.6%. As a result we determined that Mellicta ambigua can rear at natural conditions. But indoor-rearing is considered to be difficult and not useful industrially, because they have long term larval stage and only one life cycle per an year.

Home Economics teachers' concern on creativity and personality education in Home Economics classes: Based on the concerns based adoption model(CBAM) (가정과 교사의 창의.인성 교육에 대한 관심과 실행에 대한 인식 - CBAM 모형에 기초하여-)

  • Lee, In-Sook;Park, Mi-Jeong;Chae, Jung-Hyun
    • Journal of Korean Home Economics Education Association
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    • v.24 no.2
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    • pp.117-134
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    • 2012
  • The purpose of this study was to identify the stage of concern, the level of use, and the innovation configuration of Home Economics teachers regarding creativity and personality education in Home Economics(HE) classes. The survey questionnaires were sent through mails and e-mails to middle-school HE teachers in the whole country selected by systematic sampling and convenience sampling. Questionnaires of the stages of concern and the levels of use developed by Hall(1987) were used in this study. 187 data were used for the final analysis by using SPSS/window(12.0) program. The results of the study were as following: First, for the stage of concerns of HE teachers on creativity and personality education, the information stage of concerns(85.51) was the one with the highest response rate and the next high in the following order: the management stage of concerns(81.88), the awareness stage of concerns(82.15), the refocusing stage of concerns(68.80), the collaboration stage of concerns(61.97), and the consequence stage of concerns(59.76). Second, the levels of use of HE teachers on creativity and personality education was highest with the mechanical levels(level 3; 21.4%) and the next high in the following order: the orientation levels of use(level 1; 20.9%), the refinement levels(level 5; 17.1%), the non-use levels(level 0; 15.0%), the preparation levels(level 2; 10.2%), the integration levels(level 6; 5.9%), the renewal levels(level 7; 4.8%), the routine levels(level 4; 4.8%). Third, for the innovation configuration of HE teachers on creativity and personality education, more than half of the HE teachers(56.1%) mainly focused on personality education in their HE classes; 31.0% of the HE teachers performed both creativity and personality education; a small number of teachers(6.4%) focused on creativity education; the same number of teachers(6.4%) responded that they do not focus on neither of the two. Examining the level and type of performance HE teachers applied, the average score on the performance of creativity and personality education was 3.76 out of 5.00 and the mean of creativity component was 3.59 and of personality component was 3.94, higher than standard. For the creativity education, openness/sensitivity(3.97) education was performed most and the next most in the following order: problem-solving skill(3.79), curiosity/interest(3.73), critical thinking(3.63), problem-finding skill(3.61), originality(3.57), analogy(3.47), fluency/adaptability(3.46), precision(3.46), imagination(3.37), and focus/sympathy(3.37). For the personality education, the following components were performed in order from most to least: power of execution(4.07), cooperation/consideration/just(4.06), self-management skill(4.04), civic consciousness(4.04), career development ability(4.03), environment adaptability(3.95), responsibility/ownership(3.94), decision making(3.89), trust/honesty/promise(3.88), autonomy(3.86), and global competency(3.55). Regarding what makes performing creativity and personality education difficult, most HE teachers(64.71%) chose the lack of instructional materials and 40.11% of participants chose the lack of seminar and workshop opportunity. 38.5% chose the difficulty of developing an evaluation criteria or an evaluation tool while 25.67% responded that they do not know any means of performing creativity and personality education. Regarding the better way to support for creativity and personality education, the HE teachers chose in order from most to least: 'expansion of hands-on activities for students related to education on creativity and personality'(4.34), 'development of HE classroom culture putting emphasis on creativity and personality'(4.29), 'a proper curriculum on creativity and personality education that goes along with students' developmental stages'(4.27), 'securing enough human resource and number of professors who will conduct creativity and personality education'(4.21), 'establishment of the concept and value of the education on creativity and personality'(4.09), and 'educational promotion on creativity and personality education supported by local communities and companies'(3.94).

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