HUMAN PMNL MEMBRANE MICRODOMAINS AND THEIR MODIFICATIONS BY DEHP

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。先前使用Laurdan标记的PMNL悬浮液的稳态荧光的工作表明，塑料软化剂DEHP增加了PMNL膜的流动性。特别是，该化合物降低了更刚性的膜结构域的有序参数。这项研究的目的是用双光子显微镜观察单个细胞上的膜流动性区域，然后研究塑料软化剂DEHP对它们的影响。PMNL将通过Percoll梯度离心法从人血中纯化，与DEHP孵育或不与DEHP孵育，允许沉淀在玻璃片上，用Laurdan标记，并在37℃下用双光子显微镜分析Laurdan GP。

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. Previous work using steady-state fluorescence of Laurdan-labelled PMNL suspensions showed that the plastic softener DEHP increases PMNL membrane fluidity. In particular, the compound decreases the order parameter of the more rigid membrane domains. The aim of this study is to visualize membrane fluidity domains on single cells by using two-photon microscopy and then investigate how the plastic softener DEHP may affect them. PMNL will be purified from human blood by Percoll gradient centrifugation, incubated or not with DEHP, allowed to settle onto a glass coverslip, labelled with Laurdan and analysed for Laurdan GP by two-photon microscopy at 37¿C.