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• 批准号: 13671864
• 负责人: HORI Tetsuo
• 金额: $ 1.86万
• 依托单位: University of Tsukuba
• 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 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671864/
• 关键词: Congenital anal atresia; anorectal anomaly; polygenic disease; linkage mapping; pig, animal model; Anorectal malformations; anorectal anomaly; polygenic disease; linkage mapping; pig; animal model

METHODS : A pig pedigree with a high incidence of AA has been established by selective breeding using three probands from the Landrace and Large White breeds. It has been maintained by intra-familial crossing for more than fifteen years. A backcross pedigree has now been generated by mating four AA females to an unaffected male from the Chinese Meishan breed. F_1 animals were both intercrossed and backcrossed to affected AA animals. A genome scan was carried out using the F_0, F_1 and affected backcross progeny. Ninety-two microsatellite loci were analyzed using fluorescently labelled primers and an ABI377 sequencer. Linkage analysis was done with the CRI-MAP 2.4 software.RESULTS : Crossing affected parents increased the incidence of abnormalities from 30% to 61.9%. All 39 F_1 pigs were unaffected. In the F_1 intercross, only 3/205 (1.5%) were affected while 42/523 (8.0%) backcross progeny were affected. The marked difference in the incidence of affected progeny in the F_1 intercross a … More nd in the backcross indicates the presence of multiple genes causing AA. The genome scan revealed suggestive evidence for the presence of a susceptibility locus on pig chromosome 15 (lod score 2.7 for a pig microsatellite marker SW2072).CONCLUSIONS : The results clearly show that AA has a oligogenic or polygenic background. The genome scan revealed one suggestive locus causing AA on pig chromosome 15. A useful approach will also be to include markers closely linked to candidate genes for anal atresia such as genes in the Sonic Hedgehog (Shh) signalling pathway. For instance, mice that are homozygous for a targeted disruption of Gli2 encoding an Shh-responsive transcription factor show imperforate anus and recto-urethral fistula. Interestingly, GLI2 maps to human chromosome 2q14 and comparative map data suggest that the porcine homolog is likely to be located on 15q13, the region showing suggestive linkage to AA. We are currently investigating the chromosomal localization of GLI2 in pigs to tests if it is a positional candidate gene and if so it will be further evaluated in relation to the development of AA in pigs. Less

方法：利用来自长白猪和大白猪品种的三个先证者进行选育，建立了AA高发病率的猪系谱。它通过家族内部交叉维持了十五年多。现在，通过将四只 AA 雌性与来自中国梅山品种的未受影响的雄性交配，产生了回交谱系。 F_1动物与受影响的AA动物进行杂交和回交。使用F_0、F_1和受影响的回交后代进行基因组扫描。使用荧光标记引物和 ABI377 测序仪分析了 92 个微卫星位点。使用 CRI-MAP 2.4 软件进行连锁分析。结果：受影响父母的杂交使异常发生率从 30% 增加到 61.9%。所有 39 头 F_1 猪均未受影响。在 F_1 杂交中，只有 3/205 (1.5%) 受到影响，而 42/523 (8.0%) 回交后代受到影响。 F_1 杂交和回交中受影响后代的发生率存在显着差异，表明存在导致 AA 的多个基因。基因组扫描揭示了猪 15 号染色体上存在易感位点的提示性证据（猪微卫星标记 SW2072 的 lod 评分为 2.7）。 结论：结果清楚地表明 AA 具有寡基因或多基因背景。基因组扫描揭示了猪 15 号染色体上一个导致 AA 的暗示位点。一种有用的方法还包括与肛门闭锁候选基因密切相关的标记，例如 Sonic Hedgehog (Shh) 信号通路中的基因。例如，编码 Shh 响应转录因子的 Gli2 被靶向破坏的纯合小鼠表现出肛门闭锁和直肠尿道瘘。有趣的是，GLI2 映射到人类染色体 2q14，比较图谱数据表明猪同源物可能位于 15q13，该区域显示出与 AA 的暗示连锁。我们目前正在研究 GLI2 在猪中的染色体定位，以测试它是否是位置候选基因，如果是，将进一步评估其与猪 AA 发育的关系。较少的

项目成果

Hori T, Giuffra E, Andersson L, Ohkawa H: "Mapping loci causing susceptibility to anal atresia in pigs, using a resource pedigree"Journal of Pediatric Surgery. 36. 1370-1374 (2001)

Hori T、Giuffra E、Andersson L、Ohkawa H：“使用资源谱系绘制导致猪肛门闭锁易感性的位点”《小儿外科杂志》。

Hori T , eds. by Bessho F, et al.: "Anorectal malformations and Hirschsprung disease, Pathophysiology in pediatric disease"Bunkodo. 208-230 (2003)

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