• Title/Summary/Keyword: normal embryo

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The Immunohistochemical Expression of Collagens and the Morphogenesis in the Developing Mandible of Human Embryos and Fetuses (배자와 태아에서 하악골의 형태발생 및 교원질 발현에 관한 면역조직화학적 연구)

  • Kook, Yoon-Ah;Kim, Sang-Cheol;Kim, Eun-Cheol;Kim, Oh-Hwan;Kim, Jong-Ghee
    • The korean journal of orthodontics
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    • v.26 no.2 s.55
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    • pp.187-196
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    • 1996
  • Underlying malocclusions and dentofacial deformities are often related to variations in the craniofacial development. Type I and type II collagens are considered the major collagens of bone and cartilage respectively. Monitoring the patterns of those protein expressions during development will Provide a basis for the understanding of normal and abnormal growths. This study was undertaken to investigate the morphogenetic changes and the expression patterns of type I and II collagen proteins involved in the developing mandible of human embryos and fetuses. 50 embryos and fetuses were studied with Hematoxylin and Eosin, Alcian, blue-PAS, Masson Trichrome, md Immunohistochemical stains. The results were as follows : 1. A 13.5 mm embryo showed the stomatodeum with dental lamina, maxillary and mandibular processes. Meckel's cartilage appeared in the mandibular arch of a 20.5 mm embryo. New bone formation was bilaterally initiated at the outer side of middle portion of Meckel's cartilage of 22-38 mm embryos. 2. Meckel'cartilage was resorbed at the 15th week fetus. The endochondral ossification was observed where there was direct replacement of cartilage by bone. Meckel'cartilage disappeared and membraneous ossification were observed at the 25th week. 3. Before the appearance of Meckel's cartilage, the expression of type I collagen was moderate at the odontogenic epithelium of maxillary & mandibular process, but mild for the expression of type II collagen. 4. During the appearance of Meckel's cartilage and new bone formation, the immunoactivity of type II collagen was more expressed than type I collagen at the Meckel's cartilage and new bone. 5. During intrarmembranous bone formation, the expression of type II collagen was rare in the bony trabeculae. There was a switch for the expression of collagens from type II to type I during the appearance of Meckel's cartilage.

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Effect of Tris, Sodium Bicarbonate and Caffeine in Fertilization Medium on In Vitro Fertilizability of Boar Spermatozoa Frozen in Straws

  • Lee, Eun-Song
    • Reproductive and Developmental Biology
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    • v.32 no.4
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    • pp.237-243
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    • 2008
  • The objective of this study was to examine the effect of caffeine and sodium bicarbonate in a fertilization medium on the fertilizability of boar spermatozoa that were frozen in straws. Boar spermatozoa were extended with Beltsville F5 extender and frozen in 0.25-ml straws. In vitro matured porcine oocytes were fertilized in vitro (IVF) with frozen-thawed boar spermatozoa for 6h in a modified tris-buffered medium (mTBM) or in its modified medium by substituting the tris with 25mM sodium bicarbonate (modified bicarbonate-buffered medium; mBBM). Some of inseminated oocytes were fixed and stained for examination of sperm penetration. IVF embryos were cultured in a North Carolina State University-23 medium for embryo development. The percentage of live sperm was $47{\pm}4%$ and morphological abnormality of acrosome was found in $14{\pm}3%$ of spermatozoa. Optimal sperm concentration for IVF was $0.75{\sim}1.0{\times}1.0{\times}10^6$ sperms/ml when mTBM containing 5mM caffeine was used as the fertilization medium. Sperm penetration was significantly (p<0.05) stimulated by increasing caffeine concentration in the IVF medium. In addition, mBBM significantly (p<0.05) increased sperm penetration (92%) compared to mTBM (65%). More (p<0.05) blastocysts (22% vs. 32%) developed from the oocytes that were fertilized in mBBM containing 1mM caffeine than from those fertilized in mTBM with 5mM caffeine. Our results indicate that boar spermatozoa can be frozen successfully in straws with holding their normal fertilizability and that caffeine and sodium bicarbonate stimulates sperm penetration in vitro.

Successful In Vitro Development of Preantral Follicles Isolated from Vitrified Mouse Whole Ovaries

  • Kim, Dong-Hoon;No, Jin-Gu;Park, Jong-Ju;Park, Jin-Ki;Yoo, Jae Gyu
    • Reproductive and Developmental Biology
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    • v.36 no.4
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    • pp.255-260
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    • 2012
  • The purpose of this study was to assess follicular viability and competence through in vitro culture of preantral follicles isolated from vitrified mouse whole ovaries. Mouse preantral follicles were enzymatically isolated from vitrified- warmed and fresh ovaries and cultured for 10 days followed by in vitro oocyte maturation. In vitro matured oocytes were fertilized and cultured to the blastocyst stage. Five minutes pre-exposure to vitrification solution of whole ovaries had significantly higher (p<0.05) oocyte survival and maturation rates than between 10 min exposure groups. Oocyte diameter was significantly smaller (p<0.05) in the 5 and 10 min exposure groups ($69.4{\pm}2.8$ and $67.8{\pm}3.1$) when compared to that of control group ($71.7{\pm}2.1$). There was no statistical significant difference in blastocyst development rates between vitrification group (8.6%) and the fresh control group (12.0%). The mean number of cells per blastocyst was significantly lower (p<0.05) in the vitrification group ($41.9{\pm}20.2$) than in the fresh control group ($55.1{\pm}22.5$). The results show that mouse oocytes within preantral follicles isolated from the vitrified whole ovaries can achieve full maturation, normal fertilization and embryo development.

Study on Status Survey in Female Infertility Patients Admitted to Korean Medicine Hospital (한방병원에 내원한 구사(求嗣) 환자의 실태 분석 및 현황 조사)

  • Kim, Nam-Hoon;Park, Seung-Hyuk;Hwang, Deok-Sang;Lee, Jin-Moo;Lee, Chang-Hoon;Lee, Kyung-Sub;Jang, Jun-Bock
    • The Journal of Korean Obstetrics and Gynecology
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    • v.26 no.2
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    • pp.120-137
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    • 2013
  • Purpose: This study was to investigate status survey and cost of infertility patients in ${\bigcirc}{\bigcirc}$ Korean Medicine Hospital. Methods: From January 2012 to June 2012, 171 new patients were admitted to ${\bigcirc}{\bigcirc}$ Korean Medicine Hospital for treatment of infertility. We reviewed their medical records retrospectively and had telephone consultations to find out whether they were pregnant or not. Results: The mean age of outpatients was $32.72{\pm}4.08$ years and mean BMI of outpatients was $20.26{\pm}2.68$. 55.0% of patients who did not have childbirth or miscarriage. 82.5% of patients had normal menstrual period. 93.5% of patients visited OB/GYN, 33.3% were treated with ovulation induction, 18.7% underwent intrauterine insemination, and 18.1% underwent in vitro fertilization and embryo transfer. 99.7% were treated with acupuncture and moxibustion, 100% had taken Herbal Medicine. The mean treatment duration of outpatients was $8.84{\pm}8.17$ weeks, and the mean medical expense was $761,994{\pm}586,502$ won. It was found that 32.4% of patients were pregnant after treatment. Conclusions: We investigated status survey and cost of infertility patients. Further study about Korean medical treatments on infertility is required.

Hybridization Using a New Male-sterile Germplasm as the Female Parent in Chinese Jujube

  • Wang, Jiu-rui;Cui, Xiu-mei;Dai, Li;Liu, Ping;Zhao, Jin;Liu, Meng-jun
    • Horticultural Science & Technology
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    • v.33 no.3
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    • pp.396-402
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    • 2015
  • This study identified a new male-sterile germplasm of Chinese jujube, named male-sterile No. 2 (JMS2), and achieved controlled hybridization using that germplasm as the female parent. The anthers of JMS2 before flower bud opening became shrunken, dingy yellow and much smaller than normal ones, and they changed to brown after anthesis. No pollen was observed in anthers of JMS2 and its male-sterile trait remained stable over different years. A total of 1,642 fruits were obtained from ten intra- and interspecific cross combinations via controlled hybridization from 2008 to 2012 using JMS2 as the female parent. Of those, 27.3% produced seeds, on average (0-72.6%). The rate of fruit with seed (RFS) was significantly different between cross combinations or years. Compared to other cross combinations, the RFS in the combination of JMS2 ${\times}$ 'Xingguang' (a Chinese jujube cultivar with high resistance to jujube witches' broom disease) and JMS2 ${\times}$ 'Xing16' (a sour jujube genotype) remained high in different years and reached means of 48.7 and 58.1%, respectively. Finally, 150 plantlets were regenerated from immature embryos, and 51 of them were randomly selected and identified to be authentic hybrids using amplified fragment length polymorphism (AFLP) markers. This is the first report of hybrids obtained from a cross between Chinese jujube and sour jujube.

Seasonal Changes in Concentrations of Proteins and Lipids in Growing Goat Oocytes

  • Sangha, G.K.;Bhatia, H.;Khera, K.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.1
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    • pp.36-40
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    • 2007
  • Proteins and lipids not only provide a source of energy to the cell, but also play vital roles in modifying the physical properties and function of the biological membranes. In the present study, we investigated the biochemical constituents, viz. proteins and lipids, in growing oocytes of goat antral follicles during summer and winter seasons. Goat genitalia in phosphate buffered saline (pH 7.4) were brought to the laboratory within one hour of slaughter under aseptic conditions at $37^{\circ}C$. Oocytes were aspirated from normal small (<3 mm in diameter) and large (>3 mm) follicles and pooled for biochemical estimations. A significant increase in the amount of protein and lipid was observed with the growth of the oocyte. The amount of protein varied non-significantly with the season, while the amount of lipid varied significantly. The amounts of phospholipid, cholesterol, free fatty acid, and triglyceride increased with the growth of the oocyte, but no significant effect of season in these constituents was observed. Lysolecithin, sphingomyelin, and sterols were the polar lipids identified in both oocytes prepared from small follicles (small oocytes) as well as large follicles (large oocytes). In addition, the small oocytes also contained phosphatidyl serine, while large oocytes contained phosphatidyl glycerol phosphate and phosphatidyl inositol. Among non-polar lipids, triglycerides and long chain alcohols appear only in small oocytes and not in large oocytes. Monoglycerides, 1,2-diglycerides, 1,3-diglycerides and o-dialkyl glycerol ethers, fatty acids, fatty acid methyl esters, and wax esters were identified in both small and large oocytes. Information on biochemical composition of growing oocytes is relevant to oocyte and embryo competence, culture and cryopreservation.

Studies on the Improvement of Performance and Reproductive Efficiency in Farm Animals II. Study on vitro fertilization with in vivo capacitated sperm and embryo transfer in rabbits (가축의 개량 및 번식효율 증진에 관한 연구 II. 토끼에서 체내 수정능 획득정자에 의한 체외수정 및 수정란의 이식에 관한 연구)

  • 정영채;김창근;윤종택;방명걸
    • Korean Journal of Animal Reproduction
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    • v.10 no.2
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    • pp.204-210
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    • 1986
  • As a part of in vitro fertilization(IVF) for farm animals, IVF experiment was conducted using New Zealand white rabbits with their sperm capacitated in vivo. The effect of uterine conditions on sperm capacitation and effect of sperm concentration and fertilization media on IVF rate and implantation of in vitro fertilized ova were studied. The results obtained are summarized as follows; 1. Acrosomal reaction, noted after staining, of sperm recovered from ligated and intact uterus of capacitators was 83.0% and 65.7%, respectively. 2. IVF rate of ova inseminated with sperm from ligated uterus tended to be higher in DM or with higher concentration of sperm than in the modified F12 medium or with lower sperm concentration. Cleavage rate of fertilized ova for 48hr in DM was 31.5% for 106/ml and 30.0% for 104/ml of sperm and that in modified F12 medium was 26.0% for 106/ml and 22.3% for 104/ml of sperm. 3. Using the sperm from intact uterus, cleavage rate of fertilized ova showed same tendency as those shown with ligated uterus. The rate was 82.0% for 106/ml and 66.5% for 104/ml of sperm in DM and was 69.0% for 106/ml and 56.5% for 104/ml of sperm in the medium. 4. When normal ova up to 48hr after IVF were cultured for 4 days in either DM or modified F12 medium, ova developed to blastocyst stage showed higher rate in the groups of higher sperm concentration in the both media. The rate was 80.9% and 60.0% for 106/ml and 104/ml of sperm in DM and 91.7% and 71.4% for 106/ml and 104/ml of sperm in the modified F12 medium, respectively. 5. Rate of implantation after transfer of 4- or 8-cell embryos was 36.8%.

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Differential Expression of $PKD2$-Associated Genes in Autosomal Dominant Polycystic Kidney Disease

  • Yook, Yeon-Joo;Woo, Yu-Mi;Yang, Moon-Hee;Ko, Je-Yeong;Kim, Bo-Hye;Lee, Eun-Ji;Chang, Eun-Sun;Lee, Min-Joo;Lee, Sun-Young;Park, Jong-Hoon
    • Genomics & Informatics
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    • v.10 no.1
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    • pp.16-22
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    • 2012
  • Autosomal dominant polycystic kidney disease (ADPKD) is characterized by formation of multiple fluid-filled cysts that expand over time and destroy renal architecture. The proteins encoded by the $PKD1$ and $PKD2$ genes, mutations in which account for nearly all cases of ADPKD, may help guard against cystogenesis. Previously developed mouse models of $PKD1$ and $PKD2$ demonstrated an embryonic lethal phenotype and massive cyst formation in the kidney, indicating that $PKD1$ and $PKD2$ probably play important roles during normal renal tubular development. However, their precise role in development and the cellular mechanisms of cyst formation induced by $PKD1$ and $PKD2$ mutations are not fully understood. To address this question, we presently created $Pkd2$ knockout and $PKD2$ transgenic mouse embryo fibroblasts. We used a mouse oligonucleotide microarray to identify messenger RNAs whose expression was altered by the overexpression of the $PKD2$ or knockout of the $Pkd2$. The majority of identified mutations was involved in critical biological processes, such as metabolism, transcription, cell adhesion, cell cycle, and signal transduction. Herein, we confirmed differential expressions of several genes including aquaporin-1, according to different $PKD2$ expression levels in ADPKD mouse models, through microarray analysis. These data may be helpful in $PKD2$-related mechanisms of ADPKD pathogenesis.

Gametophytic Abortion in Heterozygotes but Not in Homozygotes: Implied Chromosome Rearrangement during T-DNA Insertion at the ASF1 Locus in Arabidopsis

  • Min, Yunsook;Frost, Jennifer M.;Choi, Yeonhee
    • Molecules and Cells
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    • v.43 no.5
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    • pp.448-458
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    • 2020
  • T-DNA insertional mutations in Arabidopsis genes have conferred huge benefits to the research community, greatly facilitating gene function analyses. However, the insertion process can cause chromosomal rearrangements. Here, we show an example of a likely rearrangement following T-DNA insertion in the Anti-Silencing Function 1B (ASF1B) gene locus on Arabidopsis chromosome 5, so that the phenotype was not relevant to the gene of interest, ASF1B. ASF1 is a histone H3/H4 chaperone involved in chromatin remodeling in the sporophyte and during reproduction. Plants that were homozygous for mutant alleles asf1a or asf1b were developmentally normal. However, following self-fertilization of double heterozygotes (ASF1A/asf1a ASF1B/asf1b, hereafter AaBb), defects were visible in both male and female gametes. Half of the AaBb and aaBb ovules displayed arrested embryo sacs with functional megaspore identity. Similarly, half of the AaBb and aaBb pollen grains showed centromere defects, resulting in pollen abortion at the bi-cellular stage of the male gametophyte. However, inheritance of the mutant allele in a given gamete did not solely determine the abortion phenotype. Introducing functional ASF1B failed to rescue the AaBb- and aaBb-mediated abortion, suggesting that heterozygosity in the ASF1B gene causes gametophytic defects, rather than the loss of ASF1. The presence of reproductive defects in heterozygous mutants but not in homozygotes, and the characteristic all-or-nothing pollen viability within tetrads, were both indicative of commonly-observed T-DNA-mediated translocation activity for this allele. Our observations reinforce the importance of complementation tests in assigning gene function using reverse genetics.

Long-Term Cold Storage and Plant Regeneration of Suspension Cultured Somatic Embryos of Eleutherococcus senticosus Maxim (가시오갈피 현탁배양 체세포배의 저온장기저장 및 식물체 재분화)

  • Li, Cheng Hao;Lim, Jung-Dae;Heo, Kwon;Kim, Myong-Jo;Lee, Chan-Ok;Lee, Jae-Geun;Cui, Xue Shu;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.6
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    • pp.494-499
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    • 2004
  • A method for long-term conservation of somatic embryos of Eleutherococcus senticosos was described. Suspension cultured globular somatic embryos were successfully conservated for 36 months at $4^{\circ}C$. The embryos resumed growth within two weeks when returned to MS liquid medium containing $0.2\;mg/{\ell}$. 2,4-dichlorophenoxy acetic acid. The optimal condition for cell proliferation was achieved when somatic embryos cultured at $32^{\circ}C$ in 1/3 MS liquid medium, and about 1.2 g of embryogenic cell was induced from 150 globular embryos after 6 weeks of suspension culture. The embryogenic cells produced from these somatic embryos exhibited normal plant regeneration on auxin-free medium.