THE CYTOGENETICS LABORATORY

细胞遗传学实验室

• 批准号: 7391946
• 负责人: MARK E HASKINS
• 金额: $ 10.07万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7391946
• 关键词: CYTOGENETICS; LABORATORY

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Basic studies continue to concentrate primarily on Giemsa banded karyotyping of animals with sexual anomalies as well as syndromic disorders that suggest the presence of a chromosomal anomaly. Since the last progress report, a total of 19 cytogentic studies were performed on canine and feline cases referred to the genetics clinic or following consultations. Canine investigations included DiGeorge Syndrome, os clitoris, azoospermia, ichthyosis, a PCH-1 heterozygote, intersex, hypospadias extreme, and ataxia. A feline case of intersex was also examined. Novel disease presentations included canine soft tissue swelling, ichthyosis, ataxia, extreme joint laxity, perodontal disease, DiGeorge syndrome, cystrophic mineralization in limbs, HD and ED. Feline cases of spinal muscular atrophy, ostogenesis imperfecta, multiple fractures, cutaneous asthemia, and other connective tissue disorders were also examined. Studies include Western blotting using antibodies specific for different collagens and pulse chase assays for collagen type I function and assembly. The Western blots have been challenging as many of the antibodies are specific to species other than the dog and cat. The findings in animals with skin fragility are being summarized as part of a dermatology residency project and will be submitted for publication. Another primary role of the cytogenetic laboratory is tissue culture of models currently under investigation in the Referral Center. Affected cell lines have been added for canine MPS IIIB, MPS VI, PFK, X-linked anhydrotic ectodemal dysplaisa, epidermolysis bullosa, and lethal acrodermatitis in an effort to define cellular pathology, provide RNA and DNA for mutation analysis, and for use in future cross-correction studies. Fibroblast cultures were established from dogs with hypophosphatemia, Labrador myopathy, renal failure, possible MPS III, and pathologic fractures. Fibroblast cultures from cats were established for MPS VI, an undiagnosed lysosomal storage disease, pyruvate kinase deficiency, I-cell disease, portocaval shunt, GM2 gangliosidosis, neuromuscular disease, MPS VII, MPS I, and GM1 gangliosidosis. Methods have been developed for the use of fluorescence in situ hybridization (FISH) to define the physical location of genes on canine chromosomes. This approach uses gene probes labeled with fluorescent dyes. The labeled probes are hybridized to metaphase chromosomes and detected by fluorescence microscopy, allowing cloned genes and other markers to be mapped to specific chromosome locations. The FISH method provides a means of mapping the mutant genes involved in canine genetic diseases to their chromosomal locations, further increasing knowledge of the comparative medical genetics of these important models and enhancing their use as animal models of human genetic disease. In addition, the FISH studies add to the development of the canine genome map, increasing its value as a resource for finding new canine homologs of human genetic diseases. We have collaborated in a worldwide initiative to map the dog genome. As a part of this work, we have FISH-mapped a series of markers that serve as anchors for linkage groups on canine chromosomes. These linkage groups contain 341 mapped markers distributed over 37 canine autosomes and the X Chromosome. These gene markers simultaneously serve as anchor loci for microsatellite markers in the canine linkage map and establish the chromosomal locations of the linkage groups, allowing the canine gene regions to be lined up with the homologous regions of human and mouse chromosomes. This provides access to the rich source of information on evolutionarily conserved arrangements of genes on chromosomes in these other species, greatly facilitating attempts to locate and isolate canine genes by positional cloning methods.

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子弹和调查员（PI）可能已经从其他NIH来源获得了主要资金，因此可以在其他清晰的条目中代表。列出的机构适用于该中心，这不一定是调查员的机构。基础研究继续主要集中于具有性异常的动物的GIEMSA带核分型，以及暗示存在染色体异常的综合症疾病。 自上次进度报告以来，对遗传学诊所或咨询的犬类病例总共进行了19项细胞对研究。犬类研究包括Digeorge综合征，OS阴蒂，Azoospermia，Ichthyosis，PCH-1杂合子，双性恋，Hypospadias Extreme和Hataxia。还检查了猫的猫案例。 新型疾病的表现包括犬软组织肿胀，鱼质病，共济失调，极端关节松弛，牙齿牙齿疾病，挖土综合症，四肢，高清和ED中的丘脑矿化。还检查了还检查猫脊柱肌肉萎缩，骨化不完美，多发性骨折，皮肤哮喘和其他结缔组织疾病的病例。研究包括使用针对不同胶原蛋白的抗体以及用于胶原蛋白型功能和组装的脉冲追逐测定法。蛋白质印迹一直在挑战，因为许多抗体都针对狗和猫以外的物种。皮肤脆弱性动物的发现正在总结为皮肤科住院医师项目的一部分，并将提交出版。 细胞遗传学实验室的另一个主要作用是当前在转诊中心研究的模型的组织培养。已经为犬MPS IIIB，MPS VI，PFK，X连接的藻类性藻类外神经异化症，表皮骨溶解Bullosa和致命的脂肪炎添加了受影响的细胞系，以定义细胞病理学，提供RNA和DNA，用于突变分析以及用于未来的交叉贸易研究。成纤维细胞培养物是从患有低磷酸血症，拉布拉多肌病，肾衰竭，可能的MPS III和病理性骨折的狗中建立的。建立了来自猫的成纤维细胞培养物，用于MPS VI，一种未诊断的溶酶体储存疾病，丙酮酸激酶缺乏症，I-Cell疾病，Portocaval Shunt，GM2神经节病，神经肌肉疾病，MPS VII，MPS I，MPS I和GM1 Gangliosidisoiss。 已经开发了使用荧光原位杂交（FISH）来定义基因在犬染色体上的物理位置的方法。该方法使用标记为荧光染料的基因探针。标记的探针与中期染色体杂交并通过荧光显微镜检测，从而使克隆的基因和其他标记映射到特定的染色体位置。 FISH方法提供了一种将参与犬类遗传疾病的突变基因映射到其染色体位置的方法，从而进一步增加了对这些重要模型的比较医学遗传学的了解，并增强了它们作为人类遗传疾病动物模型的使用。此外，鱼类研究增加了犬类基因组图的发展，增加了其作为寻找人类遗传疾病的新犬类同源物的资源的价值。我们已经合作制定了一项全球倡议，以绘制狗的基因组。作为这项工作的一部分，我们为一系列标记物绘制了一系列标记，这些标记是犬染色体上连锁组的锚点。这些连锁组包含341个映射的标记，分布在37个犬常染色体和X染色体上。这些基因标记物同时充当犬与犬链图中微卫星标记的锚定基因座，并建立了连锁基团的染色体位置，从而使犬基因区域与人和小鼠染色体的同源区域排成一列。这提供了有关这些其他物种中染色体上进化保守基因的丰富信息来源的访问，从而极大地促进了通过位置克隆方法定位和分离犬基因的尝试。