IDENTIFYING NEWT GENES THAT REGULATE CELLULAR PLASTICITY

鉴定调节细胞可塑性的新基因

• 批准号: 6529796
• 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/6529796
• 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）： 蝾螈具有再生多种解剖结构的非凡能力 和器官，包括四肢、脊髓、心脏、尾巴、眼睛晶状体， 视网膜、上颌和下颌。 这些再生过程依赖于 当附近完全分化的细胞去分化时 截肢部位。 这种程度的细胞可塑性是独一无二的 具有显着再生能力的生物体，在 哺乳动物。 然而，我们的实验室最近证明，最终- 分化的小鼠肌管可被诱导去分化 用再生蝾螈四肢的蛋白质提取物进行刺激。 这些结果 表明细胞去分化的信号通路是完整的 启动去分化的细胞外信号。 此举的目标 提议是鉴定启动去分化的蝾螈基因 控制哺乳动物细胞的细胞可塑性。 候选细胞可塑性 将通过差异显示分析来鉴定基因 抑制早期肢体再生和 不可再生的肢体组织。 序列分析、诱导程度和 细胞表达模式将用作选择候选者的基础 基因有待进一步研究。 表现出显着诱导和 含有的序列表明它们编码分泌蛋白，例如生长 将测试因子、细胞因子或其他配体的能力 通过用以下物质处理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 在下面的残肢组织中表达，其中包含提示的序列 它们编码细胞蛋白质，例如受体、激酶或转录 因素可能是对异位表达作出反应的基因 小鼠肌管。 使用这些方法，我们期望识别蝾螈基因 调节哺乳动物细胞的细胞可塑性。 识别 这些基因将对再生的未来产生重要影响 药品。