INTEGRIN LIGANDS IN SYNAPTIC PLASTICITY

突触可塑性中的整合素配体

• 批准号: 7336121
• 负责人: Richard Gordon LeBaron
• 金额: $ 12.5万
• 依托单位: UNIVERSITY OF TEXAS SAN ANTONIO
• 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/7336121
• 关键词: INTEGRIN; LIGANDS; SYNAPTIC; PLASTICITY

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. A. Specific Aims The scopes of the Specific Aims have not changed. B. Studies and ResultsA key approach of the work is the use of a synthetic peptide that binds integrins. The peptide was designed to contain two sequences that are recognized by commercially available antibodies. One antigen site is a six-histidine sequence (6xHis) and the second is a Myc sequence. The peptide has been made and we have linked it to a photoactive cross-linker named APDP. Using the RGD-His-Myc-APDP cross-linker (hereafter referred to as RGD-linker) a set of experiments tested whether the RGD-linker binds to hippocampus and whether the cross-linking activity was induced by light. Three activation paradigms were tested using different lights. The best source of activating light was determined to be 365 nm from a lamp that was placed approximately 6 cm from the hippocampal slice. Our first analysis of immunoblots indicate the cross-linking reaction worked. Using anti-His antibody and anti-Myc antibody several bands were detected. The overall interpretation of these experiments is that the RGD-linker functions as predicted, targeting to integrins in order to position the cross-linking activity at the integrin itself. The next set of experiments will be conducted to test whether the same bands are seen in pull-sown experiments, then a set of experiments to test whether the same molecules are cross-linked in CA1. The longer-range experiments are to identify by mass spectrometry isolated cross-linked molecules from CA1 and then knockdown their expression in organotypic cultured slice. Work on the development of organotypic culture has begun. Hippocampal slice has been placed in an incubator to test for the time it can be maintained. Supplements: Supplements have not yet been submitted. C. Significance The current significance is the potential of RGD peptide to deliver cross-linker to integrins. The preliminary results indicate that the approach is exceptionally clean and specific. This means there is little non-specific binding of RGD-linker in the tissue and could represent data showing that RGD peptide serves as an excellent targeting molecule to cells expressing integrins on their cell surface.

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。答：具体目的是特定目标的范围没有改变。 B.研究和结果的关键方法是使用结合整合素的合成肽。该肽被设计为包含通过市售抗体识别的两个序列。一个抗原位点是一个六速数序列（6xhis），第二个是myc序列。肽已经制成，我们将其与名为APDP的光活动交联。使用RGD-HIS-MYC-APDP交叉链链链（以下称为RGD链链）一组实验测试了RGD链链是否与海马结合以及是否由光引起的交联活性。使用不同的灯测试了三个激活范例。最佳的激活光源被确定为距离海马切片约6厘米的灯的365 nm。我们对免疫印迹的首次分析表明交联反应起作用。使用抗HIS抗体和抗MYC抗体，检测到了几个条带。这些实验的总体解释是RGD链链函数的预测功能，靶向整联蛋白，以将交联活性定位在整联蛋白本身。将进行下一组实验，以测试在拉链实验中是否看到相同的谱带，然后进行一组实验，以测试在CA1中是否将相同的分子交联。长期实验是通过质谱法分离的交联分子从CA1鉴定，然后敲除它们在器官培养的切片中的表达。在器官培养的发展方面的工作已经开始。海马切片已放在孵化器中，以测试可以维持的时间。 补品：尚未提交补品。 C.显着性当前的显着性是RGD肽向整联蛋白传递交联的潜力。初步结果表明该方法非常干净且具体。这意味着组织中RGD链链链几乎没有非特异性结合，并且可以表示数据表明RGD肽是对表达整联蛋白在其细胞表面表达整联蛋白的细胞的极好靶向分子。