BIOMEDICAL RESEARCH SUPPORT

该提案要求为购买光学和电子设备提供资金。 在孤立的环境中执行双波长荧光测量的设备 单细胞、小的多细胞制剂和结晶酶。 这些设备将由六名项目负责人(均为NIH现役人员)共享 在加州大学河滨分校的三个系中)。 这些项目涉及心血管疾病的一般领域。 疾病、囊性纤维化、糖尿病、钙稳态内分泌学(例如 维生素D和甲状旁腺素)、毒理学、神经肌肉传递和分子 酶催化的几个方面。对以下项目进行了描述。 Bers和Briggs博士将研究细胞质Ca(和NaI&)的调节 Phi)和钙电流。 支持收缩的钙来源(SR和Ca内流)以及 致心律失常部位的定位。 Luben博士将研究细胞质钙在悬浮液中的调节 甲状旁腺激素(PTH)处理的骨细胞和单个骨细胞 (研究亚细胞内钙的分布)以及在转染亚细胞内 甲状旁腺素受体。 Norman和Nemere博士将确定CaI的亚细胞定位 (使用Fura-2)和β-微管蛋白(使用荧光抗体) 肠上皮细胞作为维生素D的功能状态。 Quinton博士将测量[Ca]i、Phi和电生理的变化 离体汗腺对等渗吸收负荷的响应参数 来自正常和囊性纤维化受试者的导管。 亚当斯博士将使用荧光钙指示剂通过 其中一种新的蜘蛛毒素omega-agatoxin抑制细胞内钙内流 神经肌肉接头处的突触前神经末梢。他还将 探讨神经肽原球蛋白介导钙离子内流的途径 骨骼肌。 邓恩博士将研究酶和辅因子之间的分子相互作用 在四种酶的反应序列的不同步骤中 结构(晶体)状态。偏振和光谱的变化 本课程将测量固有荧光辅因子的性质。

This proposal requests funds for the purchase of optical and electronic equipment to perform dual-wavelength fluorescence measurements in isolated single cells, small multicellular preparations and crystalline enzymes. The equipment will be shared by six Project Leaders (all with active NIH grants) in three departments at the University of California, Riverside. The projects are studies involved in general areas of cardiovascular disease, cystic fibrosis, diabetes, endocrinology of Ca homeostasis (e.g. Vitamin D & PTH), toxicology, neuromuscular transmission and molecular aspects of enzyme catalysis. The following projects are described. Drs. Bers & Briggs will study the regulation of cytoplasmic Ca (and Nai & pHi) and Ca-currents in isolated cardiac myocytes, with respect to the sources of Ca which support contraction (SR & Ca influx) and also localization of arrhythmogenic sites. Dr. Luben will study the regulation of cytoplasmic Ca in suspensions of bone cells treated with parathyroid hormone (PTH) and in single bone cells (to study subcellular Ca distribution) and in cells transfected with the PTH receptor. Drs. Norman and Nemere will determine the subcellular localization of Cai (using fura-2) and beta-tubulin (using fluorescent antibodies) in isolated intestinal epithelial cells as functions of vitamin D status. Dr. Quinton will measure changes in [Ca]i, pHi and electrophysiological parameters in response to isotonic absorptive loads in isolated sweat gland ducts from both normal and cystic fibrosis subjects. Dr. Adams will use fluorescent Ca indicators to study the mechanism by which a novel spider toxin, omega-agatoxin inhibits Ca influx in presynaptic nerve terminals at neuromuscular junctions. He will also examine the means by which the neuropeptide proctolin mediates Ca entry in skeletal muscle. Dr. Dunn will study molecular interactions between enzymes and cofactors during different steps in the reaction sequence of four enzymes in structured (crystalline) states. The changes in polarization and spectral properties of intrinsically fluorescence cofactors will be measured.