Description): The study of calcium signaling in single cells is dramatically increasing our knowledge of cellular physiology. A wealth of information has been gathered on the mechanisms underlying the calcium signal and its role in cellular signal transduction. Recent technological innovations in single-cell measurements make it possible to expand this knowledge base by direct testing of many fundamental hypotheses of cell signaling. Microanalytical chemical separations enable a spatially and temporally defined quantitative measurement of key enzymes and second messengers in signal networks. Imaging of GFP-tagged proteins provides real-time information on changes in localization of many of these molecules. The present work will combine these technologies with the monitoring of intracellular calcium concentrations for in vivo studies of the activation of calcium-dependent enzymes, and of the changes of inositol 1,4,5-trisphosphate during the Xenopus laevis fertilization calcium wave.
The specific aims are 1) to measure the spatial and temporal relationship of the activation of calcium-sensing enzymes in vivo to the fertilization calcium wave, and 2) to identify the mechanisms underlying the propagation of the fertilization calcium wave.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM057015-07
Application #
6655529
Study Section
Cell Development and Function Integrated Review Group (CDF)
Program Officer
Shapiro, Bert I
Project Start
1997-09-30
Project End
2005-08-31
Budget Start
2003-09-01
Budget End
2005-08-31
Support Year
7
Fiscal Year
2003
Total Cost
$205,547
Indirect Cost
Name
University of California Irvine
Department
Physiology
Type
Schools of Medicine
DUNS #
046705849
City
Irvine
State
CA
Country
United States
Zip Code
92697
Sims, Christopher E; Luzzi, Veronica; Allbritton, Nancy L (2006) Localized sampling, electrophoresis, and biosensor analysis of Xenopus laevis cytoplasm for subcellular biochemical assays. Methods Mol Biol 322:413-24
Soughayer, Joseph S; Wang, Yan; Li, Huaina et al. (2004) Characterization of TAT-mediated transport of detachable kinase substrates. Biochemistry 43:8528-40
Hu, Shuwen; Ren, Xueqin; Bachman, Mark et al. (2004) Tailoring the surface properties of poly(dimethylsiloxane) microfluidic devices. Langmuir 20:5569-74
Wagner, John; Fall, Christopher P; Hong, Feng et al. (2004) A wave of IP3 production accompanies the fertilization Ca2+ wave in the egg of the frog, Xenopus laevis: theoretical and experimental support. Cell Calcium 35:433-47
Meredith, Gavin D; Wu, Hayley Y; Allbritton, Nancy L (2004) Targeted protein functionalization using His-tags. Bioconjug Chem 15:969-82
Hu, Shuwen; Ren, Xueqin; Bachman, Mark et al. (2003) Cross-linked coatings for electrophoretic separations in poly(dimethylsiloxane) microchannels. Electrophoresis 24:3679-88
Sims, Christopher E; Allbritton, Nancy L (2003) Single-cell kinase assays: opening a window onto cell behavior. Curr Opin Biotechnol 14:23-8
Wang, Yan; Hu, Shuwen; Li, Huaina et al. (2003) Separation of mixtures of acidic and basic peptides at neutral pH. J Chromatogr A 1004:61-70
Hu, Shuwen; Ren, Xueqin; Bachman, Mark et al. (2002) Surface modification of poly(dimethylsiloxane) microfluidic devices by ultraviolet polymer grafting. Anal Chem 74:4117-23
Wu, H; Smyth, J; Luzzi, V et al. (2001) Sperm factor induces intracellular free calcium oscillations by stimulating the phosphoinositide pathway. Biol Reprod 64:1338-49

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