Differential Manifestation of Ocular Phenotypes in TALEN-mediated p19arf Knockout FVB/N and C57BL/6J mouse lines

Jin‑Sung Park,Joo‑Il Kim,Hyun‑Jin Lim,Soo‑Kyung Ryu,Euna Kwon,Kang‑Min Han,Ki‑Taek Nam,Han‑Woong Lee,Byeong‑Cheol Kang 한국유전학회 2020 Genes & Genomics Vol.42 No.9

Background p19arf, primarily known as a tumor suppressor, has also been reported to play an essential role in normal development of mouse eyes. Consistently, lack of p19arf has been associated with ocular defects, but the mixed background of the knockout (KO) mouse strain used raised a concern on the accuracy of the phenotypes observed in association with the targeted gene due to genetic heterogeneity. Object We carried out a study to investigate into the efect of genetic background on the manifestation of p19arf KO associated phenotypes. Methods We characterized the phenotypes of novel p19arf KO mouse lines generated in FVB/N and C57BL/6J using a transcription activator-like efector nuclease (TALEN) system in comparison to the reported phenotypes of three other p19arf-defcient mouse lines generated using homologous recombination. Results Ninety-fve percent of FVB/N-p19arf KO mice showed ocular opacity from week 4 after birth which worsened rapidly until week 6, while such abnormality was absent in C57BL/6J-p19arf KO mice up to the age of 26 weeks. Histopathological analysis revealed retrolental masses and dysplasia in the retinal layer in FVB/N-p19arf KO mice from week 4. Besides these, both strains developed normally from birth to week 26 without increased tumorigenesis except for a subcutaneous tumor found in a C57BL/6J-p19arf KO mouse. Conclusion Our fndings demonstrated surprisingly variable manifestation of p19arf-linked phenotypes between FVB/N and C57BL/6J mice, and furthermore between our mouse lines and the established lines, indicating a critical impact of genetic background on functional study of genes using gene targeting strategies in mice.

스트렙토조토신 유발 당뇨병 백서에서 실험적 치아이동중의 치조골 교체

이기수,이택우,김성진 대한치과교정학회 2001 대한치과교정학회지 Vol.31 No.3

당뇨병 흰쥐를 대상으로 치조골의 교체(turnover)와 치아 이동 중 치조골의 흡수와 형성의 활성을 비교 관찰하기 위하여 8주령의 웅성 Sprague-Dawley계 흰쥐를 정상대조군(N), 당뇨군(D), 정상-치아이동군(N-tm), 당뇨-치아이동군(D-tm)으로 나누고, 각 실험군은 치아이동 실험 1일, 3일, 7일, 14일의 소군으로 나누었다. 당뇨병은 치아이동 실험 18일전에 스트렙토조토신(50mg/체중 kg)을 꼬리정맥에 단 회 주사하여 유발하였다. 치아이동은 상악 제1대구치를 초기 교정력 40그램으로 근심 이동시켰다. 상악 제1대구치 주위 치조골을 대상으로 골형성 징후표지로서 치조골 alkaline phos-phatase(ALP)와 osteocalcin(OC)을, 골흡수 징후표지로서 acid phosphatase(ACP)와tartrate-resistant acid phophatase(TRAP)을 정량 분석하였으며, D군과 N군에서는 이들의 혈중 농도를 정량 분석하였다. 1.D군의 혈청 TRAP농도와 혈청 OC농도는 모든 실험 경과군에서 N군보다 감소하였고(p<0.05), 당뇨기간이 길어질수록 더 많이 감소한 것으로 나타났다. 또한 치조골 흡수 징후표지는 감소하는 경향이었으며 골형성 징후표지도 실험기간이 경과함에 따라 더 많이 감소하는 것으로 나타났다. 2.N-tm군의 치조골 ACP농도와 TRAP농도는N군과 비교하여1일과3일군에서 가장 높았으며(p<0.01) 그 이후 감소경향을 나타냈고, 치조골 OC함량은 3일, 7일14일군에서N군보다 높았다(p<0.001), 그러나 D-tm군에서 치조골 ACP와 TRAP농도는 정상 N군에 비하여 1일에는 낮으나 7일, 14일 경과군에서 가장 높았고(p<0.01),실험 경과에 따라 계속 증가 하였으며(p<0.001), 치조골 ALP와 OC 함량은3일 경과 후부터 증가하지만 정상 N군과 유의차가 없었다. 3.당뇨-치아이동군의 치아이동량은 정상-치아이동군보다 많은 것으로 나타났다. 이러한 실험 결과는 당뇨병의 전신 골격과 치조골은 골 흡수 활성이 억제되어 있으며 골형성 활성은 당뇨 지속시간이 길어질수록 더 많이 억제되는 느린 골교체를 하며, 교정력에 의한 치조골 흡수활성 이 처음 높아지는 시기는 정상치아이동시보다 늦고 더 오랬 동안 지속되며 치조골 형성활성은 낮고, 치아 이동량은 증가하며 이동중 치아동요도가 증가함을 시사한다. 주요 단어 : 스트렙토조토신 유발 당뇨병, 치아이동, 치조골 교체, 인산효소, 오스테오칼신 The purpose of this study was to investigate the alveolar bone turnover in diabetic rat, and to compare the alveolar bone turnover during tooth movement in diabetes with that in normal control. Eighty Male Sprague-Dawley strain rats(8th week) were divided into normal control(N), normal-tooth movement (N-tm), diabetes(D), and diabetes-tooth movement(D-tm) groups. Eighteen days before the start of the experiment, diabetes was induce with a single injection of streptozotocin 50mg/kg of body weight in citrate buffer as vehicle via the tail vein. Maxillary first molars of rats were moved mesially by 40 grams of the closed coil spring. Experimental animals were sacrificed after 1d, 3d, 7d, and 14d experimental period, and the alveolar bone around the maxillary first molars were assayed biochemically for acid phsophatase(ACP) and tartrate-resistant acid phosphatase (TRAP) as bone resorption markers, and alkaline phosphatase(ALP) and osteocalcin(OC) as bone formation markers. TRAP and OC concentration in serum and alveolar bone of D group were lower than those in N group, and especially OC concentration decreased more following diabetes prolonged, which showed the decreased skeletal and alveolar bone resorption and formation potential in diabetic rats. In N-tm group compared with N group, alveolar bone ACP and TRAP concentrations were highest at 1d and 3d(p<0.01), decreased after then, and showed lowest at 14d, and alveolar bone OC concentration was higher at 3d, 7d, and 14d(p<0.001) and showed a tendency of peak level at 7d. which showed the peak of concentration of bone resorption markers at Id-3d and those of bone formation markers at 7d. In D-tm group compared with N group, alveolar bone ACP and TRAP concentrations were higher at 3d, 7d and 14d(p<0.001), and tended to reach peak value at 7d and persisted through l4d, and alveolar bone ALP and OC concentration increased but not different from that of N group. The amount of tooth movement in D group were greater than that of N group at all experimental period. Those results were suggested that during diabetes, the alveolar and skeletal bone undergo low bone turnover and the more amount of tooth movement, but because the peak time of alveolar bone resorption activity was delayed and sustained in longer period of tooth movement and alveolar bone formation activity is lower than that of normal tooth movement, the periodontal space is supposed to be larger during tooth movement. ※ Key words : Streptozotocin induced diabetes, Tooth movement, Alveolar bone turnover, Phosphatases, Osteocalcin

자라 Amyda maackii BRAUT의 췌장 내분비세포에 관한 세포화학적 연구

李學仁,金鎭澤 동국대학교 경주대학 1986 論文集 Vol.5 No.-

Cellular composition of the pancreatic islets in the freshwater turtle Amyda maackii BRAUT were studied under cytochemical technique. The results obtained are as follow ; 1. Lots of blood vessels are distributed in the pancreas of the freshwater turtle and the pancreatic endocrine cells constitute the islet at the periphery of the blood vessel. 2. The pancreatic islets are not surrounded with connective tissue capsule. 3. The pancreatic endocrine cells consist of B, A_1 and A, cells and then for the most part of the pancreatic islet are composed of only B cells or A cells. 4. B cells have the AF positive secretory granule in apical portion and A_1 cells have the cytoplasmic process for various direction. 5. A lot of endocrine cells are observed in the pancreatic exocrine tissue.

메기 Parasilurus asotus L. 췌장의 췌도세포에 관한 세포화학적 연구

李海浜,金鎭澤 동국대학교 1986 論文集 Vol.25 No.-

Pancreatic islet cells of Catfish Parasilurus asotus L. were observed under light microscope. The results obtained were as follows; 1. the pancreas of Catfish consists of thronging Parenchymal tissue and pancreatic islets were located in the exocrine portion of the pancreas. 2. The pancreatic islets were surrounded by distinctive connective tissue capsul with the same morphological characteristics as principal islets of other teleosts. 3. The pancreatic islet was composed of B, A_1, A_2 cells. 4. AF-positive B cells were located in the central portion of the pancreatic islet and A_1, A_2 cells in the periphery of the pancreatic islet. 5. A_1 cells have the long cytoplasmic processes.

관절성형술 및 측두근막 이식술을 이용한 악관절 강직증의 치험례

진택현,오승환,류동목,김여갑,이상철 大韓顎顔面成形再建外科學會 1996 Maxillofacial Plastic Reconstructive Surgery Vol.18 No.1

This is a case of bony ankylosis of right temporomandibular joint with mild facial asymmetry resulted from trauma, which bring about the right condylar fracture, and corrected by interpositional arthroplasty with temporal fascia flap and coronoidectomy. The postoperative results were functionally and esthetically good, but continued osbervation is necessary to evaluation of abnormal bony growth and reankylosis.

葉成分과 土壤의 理化學的性質이 林木生長에 미치는 영향

李光洙,鄭永觀,朴南昌,崔載彩,姜眞擇 경상대학교 농과대학 부속연습림 2001 演習林硏究報告 Vol.- No.11

This study was carried out to analyze physical and chemical properties of soil and leaf analysis to estimating amount of tree growth in Chamaecyparis pisifera, Jinhae city, Gyeongnam province. The results were summarize as follows; As the results from analysis of relation of physical-chemical properties of soil and leaf component, the relationship p component inside soil and n component inside leaf showed positive correlation with determination coefficients(R^2=0.9986), and also relation of nitrogen component inside soil and component of Na_2O inside leaf showed positive correlation with determination coefficients(R^2=0.9987), but the relation of component Ca^2+ inside soil and component of Na_2O inside leaf showed negative correlation with determination coefficients(R^2=-0.9984), and also relation of p component inside soil and Fe component inside leaf showed negative correlation with determination coefficients(R^2=-0.7215). The results from analysis of relation of leaf component and tree growth, showed correlation of determination coefficient(R^2=0.4256) in height growth, determination coefficient (R^2=0.6248) in DBH growth, determination coefficient(R^2=0.6248,) in volume growth, respectively. In each tree growth of recent five years, N and P components showed positive correlation of determination coefficient(R^2=0.6170, 0.5780, respectively) in height growth, and also N and Na^+ components showed positive correlation of determination coefficient(R^2=0.6016, R^2=0.8727, respectively) in DBH growth, but K= component showed negative correlation in DBH growth As the results from analysis of stepwise regression to estimating amount of DBH growth, sequence of inputted variables entered in order of N, Na+, Mg++, ,and estimated equation was Y=44.791N-56.640Mg+0.6867Na-26.47, determination coefficients of 69%.

세포질내 정자주입법(ICSI)에 있어서 정자흡입 및 난자내 주입방법에 관한 연구

이택후,김항진,송건호,김대근,전상식,박윤규,서태광,전병균,류은경,이은숙,문진수,김광철 경북대학교 의학연구소 2000 경북대학교병원의학연구소논문집 Vol.4 No.1

Study on Method of Sperm Aspiration and Injection into an Oocyte in Intracytoplasmic Sperm Injection(ICSI) Immobilization of spermatozoa prior to intracytoplasmic sperm iniection(ICSI) sometimes results in crooked tail and this makes it difficult to aspirate sperm into an injection pipette tail first. Head-first sperm aspiration into an injection pipette avoid this problem due to the bigger size of the sperm head. The effect of head or tail-first sperm injection into an oocyte on fertilization cleavage, percentage of grade I embryos and development to blastocyst stage in ICSI program has been studied. A single living immobilized spermatozoa from oligoasthenozoospermic patient was injected into an oocyte head-first or tail-first according to the treatment. Eighteen hours after microinjection, oocytes ware inspected for survival and fertilization Fertilized oocytes with two pronuclei were cultured in 30μl drop of mHTF supplemented with 10% heat-inactivated follicular fluid(FF) at 37℃. On day 2. embryo transfer was performed with cleaved embryos. The remaining 2-8 cell stage embryos were co-cultured with BRL cells in mHTF + 10% FF for 72 hours and the developmental stage was observed. The data were analyzed by Analysis of Variance. A total of 164 oocytes from 36 cycles were assigned to earth treatment and ICSI was performed(88 head-first, tail-first). The rates of normal fertilization were 81.8% and 76.3% for head-first and tail-first, respectively. Of the fertilized oocytes, the percentage of cleaved embryos and the percentage of grade 1 embryo among cleaved embryos were 88.9% and 68.8%, 93.1% and 74.1% for head-first and tail-first, respectively. Of the 2-8 cell embryos cultured, 44.4%(16/36) and 50.0%(10/20) for head first and tail first, respectively developed to blastocyst stage. There were no differences in fertilization, cleavage, rates of grade 1 embryos, and development to blastocyst stage. In conclusion, head-first or tail-first sperm injection into an oocyte in ICSI program does not affect fertilization and subsequent embryo development to blastocyst stage in vitro.

과학계발활동 프로그램 적용이 과학성적 우수아의 과학 탐구 능력에 미치는 효과

이승택,김진국,정재구,정진우 한국초등과학교육학회 2005 초등과학교육 Vol.24 No.1

이 연구는 초등학교 과학 성적 우수아를 대상으로 과학계발활동 프로그램의 구성 및 적용이 과학탐구능력에 미치는 효과를 분석하였다. 분석결과, 과학계발활동 프로그램을 적용한 학생들은 적용하지 않은 학생들에 비하여 관찰, 측정, 자료해석, 가설 설정, 변인통제의 과학탐구요소가 더 신장된 것으로 나타났다. 따라서 과학계발활동의 운영은 과학탐구능력의 신장에 효과를 기대할 수 있으므로 교육현장에서 과학교육을 보다 다양하고 수준별로 운영할 수 있는 프로그램의 개발 및 적용 방안을 모색할 필요가 있다. This study organized a Science Improvement Activity Program designed for the excellent students in science in 5th and 6th grades. After application the effect of the program on science process skills was analyzed. The Science Improvement Activity Program was applied to the experimental group as a club activity after school during the semester. The results of this study were as follows: First, the students participated in the Science Improvement Activity showed an increase in the science process skills compared with those who didn't. Second, it was found that the students to whom the Science Improvement Activity Program was applied have improved in enhancing their science process elements of observation, measurement, data analysis, forming hypotheses, and controlling variables. It is expected that the development of activity programs and their active application should be done in order for schools to provide science education according to each student's capacity from various angles.

pHOxsFV벡터와 배아주간세포를 이용한 형질전환생쥐 생산 기초연구

이훈택,이봄이,정길생,김진회 건국대학교 동물자원연구센터 1998 動物資源硏究誌 Vol.19 No.-

pHook™-1 hapten 4-ethoxy-methylene-2-phenyl-2-oxazolin-5one(phOX)의 단일 항체 sFV를 암호화 하고 있으며, murine의 Ig k-chain V-J2-C 영역유래의 signal peptide에 의하여 항체를 세포 표면에 배열시키도록 고안되어 있다. 또한, 항체를 세포막 바깥쪽에 부착되어 있도록 하기 위해 PDGFR 유래의 transembrane domain의 C 말단에 결합되어 있다. 이렇게 고안된 vector을 발현하는 세포는 세포막에 sFV을 발현함으로, phOX로 코팅된 자석베드를 이용하여 배양체로부터 목적의 유전자를 발현하는 세포만을 분리할 수 있을 것이다. 따라서, 본 연구는 pHook™-1 유전자를 co-transfection함으로써 목적의 유전자를 가진 배아주간세포를 단시간 내에 효율적으로 선발하기 위하여 실시하였다. 또한, 배아주간세포에서 목적의 DNA 발현 또는 존재를 검증하기 위해 DNA 발현 또는 존재를 검증하기 위해 PCR 방법과 조직화학적 방법을 사용하였다. 형질전환유전자 발현을 transfection(유전자 전이) 후 4∼14일 사이에 모든 배아주간세포에서 확인되었다. Magnetic bead를 이용하여 선발된 세포에서 co-transfected DNA는 배아주간세포에서 효율절으로 삽입되었으며, 선발된 세포의 약 90%는 co-transfected 유전자를 발현하였다. 이 결과는 세포생리학에서 특이 유전자의 급성변이와 만성변이를 연구하거나, 또는 형질전환동물을 생산하기 위해 pHook™-1 목적유전자와 함께 전이함으로서 효율적으로 목적의 유전자를 가진 세포를 선발 가능함으로써 보다 간편하게 형질전환도 동물의 생산에 이용 가능하다는 사실을 확인하였다. pHook™-1 encodes a single-chain antibody(sFv) directed toward the hapten 4-ethoxy-methylene-2-phenyl-2-oxazolin-5-one(phOx): the signal peptide from the murine Igk-chain V-J2-C region is fused in front of coding region of the sFv to direct the antibody to the plasma membrane. The antibody is fused at the C-termius to the transmembrane domain from the platelet derived growth factor receptor(PDGFR), allowing the antibody to be anchored and displayed on the extracellular side of theemmbrane. Transfected cells expression sFv can be isolated from whole cultures by using magnetic coated with phOx and a strong magnetic strand. Thus, the present study was designed to apply the embryonic stem cells by using pHook™-1 . Cell-transduction efficiency was measured by morphometric analysis. Polymerase chain reaction and histochemistry were used to detect the presence and/or expression of objective DNA in embryonic stem cells. Transgene expression was detected in all cases between 4 and 14 day after transfection. In selected cells using magnetic bead, co-transfected DNA was also incorporated efficiently in embryonic stem cells and approximately 90% of the selected cells expressed co-transfected gene. This result suggested that this selection system can be used as a feasible tool, when pHook™-1 is cotransfected with objective gene, to isolate and study for acute and chronic changes of a specific gene in cellular physiology.

자궁경부의 상피내암 조직내 In Situ Hybridization에 의한 Human Papillomavirus DNA의 검색 및 Typing

이상숙,김기권,정재홍,김택훈,이태성,진승원,손우익 대한의학협회 1990 대한의학협회지 Vol.33 No.1