Using a cultured ovine lens system, Dr. Louis will pursue 6 specific aims: 1.Determine whether IP3, Ca2+ or cADPR (cyclic ADP-ribose) are responsible for propagation of cell-cell Ca2+ waves in the lens. Each of these will be microinjected into single cells in culture and determining whether the resulting cell-cell Ca2+ waves are affected by a variety of pharmacological reagents that are specific for different enzymes and channels used by 2nd messenger molecules. 2.Absolute concentrations of IP3 and cADPR will be measured following addition of agonists which elevate Ca2+. Cell surface receptors that mediate the agonist or mechanical stimulation-dependent elevation of Ca2+ will be determined. Then the PtdIP2-phospholipase C(s), ADP-ribosyl cyclase(s) will be identified. Then the channel proteins responsible for elevating Ca2+ identified. 3.Identify the agonists and enzymes responsible for cAMP signaling to determine the relationship between agonists that elevate Ca2+ and the activation of adenylate cyclase. Effects of membrane permeable activators of protein kinases, phosphatase inhibitors and calmodulin inhibitors will be studied to determine whether kinases or calmodulin affect Ca2+ waves. 4.The more differentiated lens """"""""fibers"""""""" have higher resting Ca2+. Experiments will determine if the """"""""fibers"""""""" have different Ca2+ regulatory mechanisms. 5.Effects of oxidizing agents on lens cell cytosolic Ca2+ will be determined using both membrane permeant and impermeant -SH reagents. 6.The roles of the different lens connexins in mediating the Ca2+ waves will be studied by measuring Ca2+ permeability of both homotypic and heterotypic gap junctions in HeLa cells.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
7R01EY005684-17
Application #
6384488
Study Section
Visual Sciences A Study Section (VISA)
Program Officer
Liberman, Ellen S
Project Start
1985-04-01
Project End
2004-08-31
Budget Start
2001-04-01
Budget End
2004-08-31
Support Year
17
Fiscal Year
2001
Total Cost
$221,786
Indirect Cost
Name
Georgia State University
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
837322494
City
Atlanta
State
GA
Country
United States
Zip Code
30302
Zou, Juan; Salarian, Mani; Chen, Yanyi et al. (2017) Direct visualization of interaction between calmodulin and connexin45. Biochem J 474:4035-4051
Zou, Juan; Salarian, Mani; Chen, Yanyi et al. (2014) Gap junction regulation by calmodulin. FEBS Lett 588:1430-8
Xu, Qin; Kopp, Richard F; Chen, Yanyi et al. (2012) Gating of connexin 43 gap junctions by a cytoplasmic loop calmodulin binding domain. Am J Physiol Cell Physiol 302:C1548-56
Chen, Yanyi; Zhou, Yubin; Lin, Xianming et al. (2011) Molecular interaction and functional regulation of connexin50 gap junctions by calmodulin. Biochem J 435:711-22
El-Shamayleh, Yasmine; Kiorpes, Lynne; Kohn, Adam et al. (2010) Visual motion processing by neurons in area MT of macaque monkeys with experimental amblyopia. J Neurosci 30:12198-209
Zhou, Yubin; Yang, Wei; Lurtz, Monica M et al. (2009) Calmodulin mediates the Ca2+-dependent regulation of Cx44 gap junctions. Biophys J 96:2832-48
Lurtz, Monica M; Louis, Charles F (2007) Intracellular calcium regulation of connexin43. Am J Physiol Cell Physiol 293:C1806-13
Zhou, Yubin; Yang, Wei; Lurtz, Monica M et al. (2007) Identification of the calmodulin binding domain of connexin 43. J Biol Chem 282:35005-17
TenBroek, E M; Johnson, R; Louis, C F (1994) Cell-to-cell communication in a differentiating ovine lens culture system. Invest Ophthalmol Vis Sci 35:215-28
Weaver, C D; Shomer, N H; Louis, C F et al. (1994) Nodulin 26, a nodule-specific symbiosome membrane protein from soybean, is an ion channel. J Biol Chem 269:17858-62

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