应用抑制性消减杂交技术鉴定母鼠铅暴露后代海马差异表达基因

Suppression subtractive hybridization for the identification of differentially expressed genes in the hippocampus of the offsprings of lead exposed female rats

摘要:

目的：筛查和鉴定与铅暴露相关的海马回中差异表达基因，为鉴定学习和记忆障碍相关基因提供理论基础。方法：母体铅暴露后子代学习记忆障碍的幼鼠脑海马提取RNA，采用抑制性消减杂交技术，鉴定母鼠铅暴露后代海马差异表达基因。结果：构建了1个近200个克隆的差减文库，测序表明得到有效克隆93个，并从中克隆和鉴定了母鼠铅暴露致学习和记忆能力显著下降的幼鼠中的海马中差异表达的基因，共43种不同的基因以及4种未知功能基因。结论：在铅暴露造成幼鼠海马中明显上调的基因包括一些已知的看家基因编码蛋白和一些与细胞蛋白折叠、信号传导、应激响应及DNA甲基化相关功能的蛋白。这些蛋白可能与幼鼠学习、记忆能力的显著降低直接相关。［中国当代儿科杂志，2010，12（10）：820-824］

Abstract:

OBJECTIVE: To screen and identify differentially expressed genes in the hippocampus of the offsprings of lead exposed female rats in order to provide a theoretical basis for identifying learning and memory deficits related genes. METHODS: RNA was extracted from the hippocampus of young rats with learning and memory deficits due to maternal lead exposure. Suppression subtractive hybridization was used to identify the differentially expressed genes in the hippocampus. RESULTS: An effective subtracted library was constructed which consisted of approximately 200 clones. Sequencing for the library identified 93 clones harboring insertion fragments which included 43 different genes and 4 unknown genes. These genes might be related to learning and memory deficits due to maternal lead exposure. CONCLUSIONS: The up-regulated genes in the hippocampus of young rats from pregnant rats under lead exposure include some housekeeping genes and some proteins involved in cellular protein folding, signal transduction, stress response and DNA methylation. These proteins might be directly related to a significant reduction in learning and memory abilities in the young rats. ［Chin J Contemp Pediatr, 2010, 12 (10):820-824］