IDENTIFYING NEWT GENES THAT REGULATE CELLULAR PLASTICITY

鉴定调节细胞可塑性的新基因

• 批准号: 6437214
• 负责人: SHANNON J ODELBERG
• 金额: $ 33.75万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-30 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6437214
• 关键词: Urodela; adipose tissue; cell differentiation; complementary DNA; developmental genetics; differential display technique; gene induction /repression; limb regeneration; molecular cloning; muscle cells; muscle proteins; myotubes; nucleic acid sequence; osteocytes; stem cells; subtraction hybridization

DESCRIPTION (provided by applicant): Newts have a remarkable capacity to regenerate several anatomical structures and organs including their limbs, spinal cords, heart, tails, eye lenses, retinas, and upper and lower jaws. These regenerative processes are dependent upon the dedifferentiation of fully-differentiated cells in the vicinity of the amputation site. This degree of cellular plasticity is unique to organisms with marked regenerative capabilities and is not observed in mammals. However, our laboratory has recently demonstrated that terminally- differentiated mouse myotubes can be induced to dedifferentiate when stimulated with a protein extract from regenerating newt limbs. These results indicate that the signaling pathways for cellular dedifferentiation are intact in extracellular signals that initiate dedifferentiation. The goal of this proposal is to identify the newt genes that initiate dedifferentiation and control cellular plasticity in mammalian cells. Candidate cellular plasticity genes will be identified by performing differential display analysis and suppression subtractive cDNA hybridization between early limb regenerates and non regenerating limb tissues. Sequence analysis, degree of induction, and cellular expression patterns will be used as a basis for selecting candidate genes for further study. Genes that exhibit a significant induction and contain sequences suggesting they encode secreted proteins such as growth factors, cytokines or other ligands will be tested for their ability to initiate cellular dedifferentiation by treating cultured mouse myotubes with conditioned medium from cells expressing these candidate genes. Genes expressed in the underlying stump tissue that contain sequences suggesting they encode cellular protein such as receptors, kinases or transcription factors could be genes that respond to the ectopically expressing them in mouse myotubes. Using these methods, we expect to identify newt genes that function in regulating cellular plasticity in mammalian cells. Identifying these genes would have important implication for the future of regenerative medicine.

描述（由申请人提供）： 纽特具有重生几个解剖结构的显着能力 和器官，包括四肢，脊髓，心脏，尾巴，眼镜， 视网膜以及上下颌。 这些再生过程取决于 完全分化的细胞在附近的完全分化的细胞进行推导后 截肢站点。 这种程度的细胞可塑性是独一无二的 具有明显再生能力的生物，在 哺乳动物。 但是，我们的实验室最近证明了这一点 分化的鼠标肌管可被诱导去分化 用再生的纽特四肢刺激蛋白质提取物。 这些结果 表明细胞去分化的信号传导途径完整 在启动去分化的细胞外信号中。 目标的目标 提案是确定启动去分化和的NEW基因 控制哺乳动物细胞中的细胞塑性。 候选细胞塑性 将通过进行差异显示分析和 抑制早期肢体再生之间的减法cDNA杂交 非再生肢体组织。 序列分析，诱导程度和 蜂窝表达模式将被用作选择候选者的基础 用于进一步研究的基因。 表现出显着诱导的基因 包含序列，表明它们编码分泌的蛋白质，例如生长 因素，细胞因子或其他配体的能力将测试 通过用使用培养的小鼠肌管来启动细胞去分化 来自表达这些候选基因的细胞的条件培养基。 基因 在包含序列的下面的树桩组织中表达 它们编码细胞蛋白，例如受体，激酶或转录 因素可能是对异位表达它们反应的基因 鼠标肌管。 使用这些方法，我们希望确定 在调节哺乳动物细胞中细胞可塑性的功能。 识别 这些基因将对再生的未来具有重要意义 药品。