UNIVERSITY OF MASSACHUSETTS
This is an ARRA administrative supplement to enable the Specific Aims of parent grant NIH 5 R37 GM030626 to be achieved more quickly and with greater insight. Funds have been awarded to equip an existing light microscope for Differential Interference Contrast (DIC) optics and high-speed video recording, which will allow the direct observation and documentation of the movement of intraflagellar transport (IFT) particles within the flagella of the model organism Chlamydomonas. Such observations will be critical to determining how mutations in proteins such as IFT46 and nephrocystin-4 affect the size, number, and rate of movement of IFT particles in the flagellum, and thus provide insight important for achieving Specific Aim 1 (focused on IFT46) and Specific Aim 6 (focused on nephrocystin-4). Funds also have been provided to acquire an imaging system for quantitation of protein levels in western blots using chemiluminescence. This system will reveal how the levels of specific proteins are affected in the flagella of mutants for IFT46 (Specific Aim 1) and nephrocystin-4 (Specific Aim 6). Cilia and flagella occur throughout the human body, and defects in them cause diseases such as primary ciliary dyskinesia, polycystic kidney disease, and blindness. This research is providing new insights into the basic mechanisms involved in the assembly and function of cilia and flagella, revealing how defects in disease-related proteins affect these organelles, and identifying new candidate disease genes for primary ciliary dyskinesia and other cilia-related diseases.