RESEARCH OF THE MUTATED GENE RESPONSIBLE FOR RAT OSTEOCHONDRODYSPLASIA AND INVESTIGATION OF THE EMBRYONIC PATHOGENESIS.

大鼠骨软骨发育不良突变基因的研究及胚胎发病机制的研究。

• 批准号: 13660309
• 负责人: SUZUKI Katsushi
• 金额: $ 2.3万
• 依托单位: Nippon Veterinary and Life Science University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13660309/
• 关键词: OSTEOCHONDRODYSPLASIA; LINKAGE ANALYSIS; CONGENITAL ANORMALIES; RATS; ミュータント; 第11染色体; ocd遺伝子; 胎生期病態; マイクロサテライトリンケージ; RHパネル

The gene, ocd, responsible for rat osteochondrodysplasia is located on rat chromosome 11. In this study, we made a fine map around the ocd locus to identify the candidates. We also revealed the pathogenesis of the mutant rat during late embryonic stage. Linkage analysis was performed using backcross progeny obtained from the mating between HGN (hgn/hgn) females and F1 (BNxHGN:+/hgn) males. RH map around the ocd locus was obtained by typing Rat/Hamster radiation hybrid clones with closely related markers to the ocd locus. In another linkage analysis, we genotyped the affected rats of the OCD strain with microsatellite (MS) makers showing polymorphism in the strain. The ocd locus was finally localized into 100kb-region on rat Chr.11. At present, two candidate genes for ocd are located in the region. RT-PCR analysis revealed the expressions of these genes in the fetal tissue of the affected rats. In the experiments of embryonic pathogenesis, at first, we confirmed the pleiotropic effects … More of the ocd on the backcross newborns (derived from HGN x F1) under altered genetic background, indicating that ocd affects on the fetal development of multiple organs. At second, we revealed the fetal pathogenesis of the affected rats of JCD strain. The fetal rats were obtained from pregnant mothers by cesarean section at embryonic day (ED) 16.5-21.5. They were genotyped by the PCR of the MS flanking the ocd locus. The body weight was significantly smaller in the affected than normal fetuses at ED 16.5. The reductions of the length of the limbs became severe with increase of embryonic ages. The double-staining with alcian blue and alizarin red S revealed that systemic cartilage formation of the affected fetus was already hypoplasitc at ED 16.5 and that the ossification was accelerated in the affected rat after ED 18.5. Histological examination revealed that, although the ossification center was formed, the accumulation of cartilage matrices was poor in the affected rats after ED 16.5. These results suggnst that normal allele for ocd might be expressed before ED 16.5 and that the normal expression of the gene is critical for, normal development. Less

导致大鼠骨软骨发育不良的基因ocd位于大鼠11号染色体上。在这项研究中，我们围绕强迫症基因位点制作了一个精细的图谱来识别候选者。并揭示了该突变大鼠胚胎晚期的发病机制。使用从HGN（hgn/hgn）雌性和F1（BNxHGN：+/hgn）雄性之间交配获得的回交后代进行连锁分析。用与ocd基因座密切相关的标记对大鼠/γ-射线杂交克隆进行分型，获得ocd基因座周围的RH图谱。在另一个连锁分析中，我们用微卫星（MS）标记对强迫症品系的患病大鼠进行基因分型，该标记在该品系中显示多态性。ocd位点最终定位于大鼠第11染色体100 kb区域。目前，两个强迫症的候选基因位于该区域。RT-PCR分析显示，这些基因的表达在胎儿组织中的受影响的大鼠。在胚胎发病机制的实验中，我们首先证实了多效性效应 ...更多信息 在遗传背景改变的情况下，回交后代（HGN × F1）的强迫症发生率为100%，表明强迫症影响胎儿多器官的发育。其次，揭示了JCD系病鼠的胎鼠发病机制。胎鼠在胚胎日（艾德）16.5-21.5通过剖宫产从孕鼠获得。通过PCR对位于ocd基因座侧翼的MS进行基因分型。在艾德16.5时，受影响胎仔的体重显著小于正常胎仔。随着胎龄的增加，肢长的缩短程度加重。阿辛蓝和茜素红S双重染色显示，在艾德16.5时，受累胎仔的全身软骨形成已经发育不良，在艾德18.5后，受累大鼠的骨化加速。组织学检查显示，虽然骨化中心已形成，但在艾德16.5后，受累大鼠中软骨基质的积累较差。这些结果表明，正常的ocd等位基因可能在艾德16.5岁以前表达，该基因的正常表达是正常发育的关键。少

项目成果

Suzuki H, Yagi M, Saito K, Suzuki K: "Dysplastic development of seminilerous tubules and interstitial tissue in rat hypogonadic hgn/hgn testes."Biology of Reproduction. 71(1)(In press). (2004)

Suzuki H、Yagi M、Saito K、Suzuki K：“大鼠性腺功能减退 hgn/hgn 睾丸中生精小管和间质组织的发育不良。”生殖生物学。

Suzuki H, Yagi M, Saito K, Suzuki K: "Dysplastic development of seminiferous tubules and interstitial tissue in rat hypogonaclic hgn/hgn testes."Biology of Reproduction. 71(1)(in press). (2004)

Suzuki H、Yagi M、Saito K、Suzuki K：“大鼠性腺功能低下 hgn/hgn 睾丸中曲细精管和间质组织的发育不良。”生殖生物学。

Cupp AS, Uzumcu M, Suzuki H, et al.: "Effect of transient embryonic in vivo exposure to the endocrine disruptor methoxychlor on embryonic and postnatal testis development."Journal of Andrology. 24(5). 736-745 (2003)

Cupp AS、Uzumcu M、Suzuki H 等人：“短暂胚胎体内暴露于内分泌干扰物甲氧氯对胚胎和出生后睾丸发育的影响。”男科学杂志。

Uchibori M., K.Saito, S.Yokoyaina, H.Suzuki, T.Tsuji, K.Suzuki: "Foci of spike discharges in sleeping EEG of E1 mice can be determined mathematically with wavelet transform of multiple monopolar derivations in individual animals based on the electric fiel

Uchibori M.、K.Saito、S.Yokoyaina、H.Suzuki、T.Tsuji、K.Suzuki：“E1 小鼠睡眠 EEG 中尖峰放电的焦点可以通过对个体动物中多个单极导数的小波变换进行数学确定。

Inomata, A., I.Horii, K.Suzuki: "5-Fluorouracil-induced intestinal toxicity : what determines the severity of damage to murine intestinal crypt epithelia?"Toxicology Letters. 133. 231-240 (2002)

Inomata，A.，I.Horii，K.Suzuki：“5-氟尿嘧啶诱导的肠道毒性：什么决定了小鼠肠隐窝上皮损伤的严重程度？”毒理学快报。