THE ROLE OF MURINE AIM1 IN NEURAL CREST/MELANOCYTE DEVELOPMENT

AIM 1在消减杂交中被鉴定 选择黑色素瘤中不存在的cDNA的实验 （AIM）。AIM 1在人致瘤性黑色素瘤中不表达 细胞系（UACC- 903），但在非致瘤性 通过引入人抑制的黑色素瘤细胞系 6号染色体（MCH-361 C1）。我们正在开发一种小鼠模型 系统研究AIM 1在黑色素瘤生物学和正常 黑素细胞发育AIM 1的小鼠同源物， muAIM 1，从成人肝脏cDNA文库中分离，使用 GeneTrapper（LTI）方法。核酸比较 序列和推导的开放阅读框的正确率均在85%以上 在基因的限制性区域的身份。序列数据库检索 AIM 1与β，γ-晶体蛋白相似的鉴定区域 家族，两侧是AIM 1区域，与 其他蛋白质。muAIM 1在小鼠基因组中的位置是 使用杰克逊实验室回交通过PCR-RFLP检查 panel并发现位于近端小鼠染色体10上; a 与人类染色体6同线的区域。进一步调查 AIM 1在黑素细胞发育和恶性肿瘤中的作用 转化、RT-PCR、北方印迹和原位杂交 分析用于确定muAIM 1的表达模式 在成年小鼠组织和胚胎发育期间。 与毛囊发育的相关性最为显著 发展特征

AIM1 was identified in a substractive hybridization experiment that selected for cDNAs that are absent in melanoma (AIM). AIM1 is not expressed in a human tumorigenic melanoma cell line (UACC- 903) yet highly expressed in a non-tumorogenic melanoma cell line suppressed by introduction of human chromosome 6 (MCH- 361C1). We are developing a mouse model system to study the role of AIM1 in melanoma biology and normal melanocyte development. The mouse homologue of AIM1, muAIM1, was isolated from an adult liver cDNA library using the GeneTrapper (LTI) method. Comparison of the nucleic acid sequences and deduced open reading frame demonstrated over 85% identity in restricted areas of the gene. Sequence database searches identified regions of AIM1 with similarity to beta, gamma-crystallin family, flanked by regions of AIM1 with no obvious similarity to other proteins. The location of muAIM1 in the mouse genome was examined by PCR- RFLP using the Jackson laboratory backcross panel and found to reside on proximal mouse chromsome 10; a region syntenic to human chromsome 6. To further investigate the role of AIM1 in melanocyte development and malignant transformation, RT-PCR, Northern blot and in situ hybridization analyses were used to determine the expression pattern of muAIM1 in adult mouse tissues and during embryonic development. Correlation with the developing hair follicle was the most striking developmental feature.