Perturbation of the smooth muscle cells (SMC) in the vascular wall can result in their proliferation and migration that, along with fibrous/fibrofatty accumulation contribute to intimal thickening, and result in atherosclerosis, the major cause of vascular disease. In order to understand the biology of the SMC and the physiological basis for the differences in phenotype of the medial SMCs, we propose to investigate the molecular biology and mechanics of single SMCs from rabbit smooth muscle tissue under normal and experimentally induced pathological conditions. As a prelude to understanding the cellular biology of medial SMCs that form the intimal thickening in atherosclerosis, we propose to determine: a) how variable expression of the myosin isoforms can alter the mechanical function (unloaded shortening, extent of shortening and maximal force production) of SMCs, and b) possible further modification of these properties via the second messenger pathways which are used to regulate these cells. Specifically, we propose to study the physical and mechanical properties of isolated single SMCs from the vascular (carotid, femoral, renal and saphenous), and digestive (stomach and intestine) systems. We will use single SMC RT-PCR to determine the myosin heavy (SMA/B and SMl/2) and light chain (MLC17a/b) isoform composition from the same cell on which the mechanical measurements were made in order to correlate these parameters.
The specific aims of this grant are to determine how myosin isoform expression, correlates with filament organization, and stability, and thereby alters cell mechanics in normal and pathological conditions. This entails examining: 1) Cells of known myosin isoform composition to determine how isoform expression and initial cell length affects unloaded shortening velocity, extent of shortening and force production. 2) Tissues and isolated cells using immunohistochemistry to determine if there is unique myosin isoform distribution and localization within SMC's. 3) If second messenger systems can further alter the """"""""inherent"""""""" unloaded shortening velocity in cells of known myosin isoform composition. 4) If SMC's, which are involved in the migration and proliferation resulting from experimentally induced atherosclerosis have unique myosin isoform expression and mechanical properties. ? ?

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
3R01HL062237-05A1S1
Application #
6914729
Study Section
Experimental Cardiovascular Sciences Study Section (ECS)
Program Officer
Ershow, Abby
Project Start
1999-04-01
Project End
2007-11-30
Budget Start
2003-12-15
Budget End
2004-11-30
Support Year
5
Fiscal Year
2004
Total Cost
$36,250
Indirect Cost
Name
Marquette University
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
046929621
City
Milwaukee
State
WI
Country
United States
Zip Code
53201
Eddinger, Thomas J (2014) Smooth muscle-protein translocation and tissue function. Anat Rec (Hoboken) 297:1734-46
Huang, Qian; Babu, Gopal J; Periasamy, Muthu et al. (2013) SMB myosin heavy chain knockout enhances tonic contraction and reduces the rate of force generation in ileum and stomach antrum. Am J Physiol Cell Physiol 304:C194-206
Eddinger, Thomas J (2009) Unique contractile and structural protein expression in dog ileal inner circular smooth muscle. J Smooth Muscle Res 45:217-30
Govindaraju, Sandya R; Bain, James Lw; Eddinger, Thomas J et al. (2008) Vibration causes acute vascular injury in a two-step process: vasoconstriction and vacuole disruption. Anat Rec (Hoboken) 291:999-1006
Eddinger, Thomas J; Meer, Daniel P; Miner, Amy S et al. (2007) Potent inhibition of arterial smooth muscle tonic contractions by the selective myosin II inhibitor, blebbistatin. J Pharmacol Exp Ther 320:865-70
Eddinger, Thomas J; Schiebout, Jessen D; Swartz, Darl R (2007) Adherens junction-associated protein distribution differs in smooth muscle tissue and acutely isolated cells. Am J Physiol Gastrointest Liver Physiol 292:G684-97
Eddinger, Thomas J; Meer, Daniel P (2007) Myosin II isoforms in smooth muscle: heterogeneity and function. Am J Physiol Cell Physiol 293:C493-508
Han, Shaojie; Speich, John E; Eddinger, Thomas J et al. (2006) Evidence for absence of latch-bridge formation in muscular saphenous arteries. Am J Physiol Heart Circ Physiol 291:H138-46
Call, Christopher; Han, Shaojie; Speich, John E et al. (2006) Resistance to pressure-induced dilatation in femoral but not saphenous artery: physiological role of latch? Am J Physiol Heart Circ Physiol 291:H1513-20
Eddinger, Thomas J; Schiebout, Jessen D; Swartz, Darl R (2005) Smooth muscle adherens junctions associated proteins are stable at the cell periphery during relaxation and activation. Am J Physiol Cell Physiol 289:C1379-87

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