Core B is the Cell Culture and Small Animal Core. The function of the Cell Culture component is to provide Program Investigators with complete cell culture and cell isolation services that facilitate completion of their specific aims. In this capacity, the Core will: a) achieve economy of scale and quality assurance through centralized cell and tissue processing;b) isolate cells from the transgenic mice, including tau -/- (Project 1), PDE4B -/-, and PDE4D -/- (Project 2), and alG -/-, and eNOS -/- mice (Project 3);c) provide technical expertise, consultation, and training in cell isolation and culture;and d) serve as a centralized repository and source of endothelial cells and information generated from them. Considering that endothelial cells lining the pulmonary arteries, alveolar capillaries, and pulmonary veins exhibit remarkable heterogeneity both in structure and in function, the isolation and culture of endothelial cells from these different vascular segments is essential to the success of this application. Our Core routinely isolates pulmonary artery (PAEC), microvascular (PMVEC), and vein (PVEC) endothelial cells from transgenic and non-transgenic rats and mice. These cells are made available to investigators as needed. Specifically, we provide a seeding service, so that cell requests are filled twice per week, at the requested seeding density and in the requested tissue culture format. The Core currently maintains multiple cell lines for each aforementioned phenotype, seeds "95 million cells per week, and provides cells to more than 50 laboratories nationally. The small animal component is a new constituent of the program. The function of the Small Animal component is to provide Program Investigators with a clinically relevant animal model of lung injury that facilitates completion of their Specific Aims. In this capacity, the core will: a) expand and maintain wild type and genetically modified strains of Pseudomonas aeruginosa;b) inject rats and mice with P. aeruginosa to elicit lung injury (a clinically relevant etiology of acute lung injury);c) monitor injected animals with P. aeruginosa until execution of terminal experiments;d) provide technical expertise, consultation and training in bacterial expansion and the animal model, and e) serve as a centralized repository of lung samples from infected and non-infected animals.

Public Health Relevance

Isolation of endothelial cells Is fundamental to address the existing heterogeneity in signaling events proposed in the current application. The Cell Culture component of the Core centralizes isolation of the required endothelial cells, and provides the reagents necessary for completion of the proposed experiments in all projects. P. aeruginosa is a main clinical etiology of acute lung injury. The Small Animal component of the Core centralizes the development of a lung Injury model and provides training for the assessment of outcomes measures necessary for completion of the proposed experiments in all projects.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Program Projects (P01)
Project #
2P01HL066299-11A1
Application #
8293877
Study Section
Heart, Lung, and Blood Initial Review Group (HLBP)
Project Start
Project End
Budget Start
2012-05-15
Budget End
2013-03-31
Support Year
11
Fiscal Year
2012
Total Cost
$306,392
Indirect Cost
$100,067
Name
University of South Alabama
Department
Type
DUNS #
172750234
City
Mobile
State
AL
Country
United States
Zip Code
36688
Sellak, Hassan; Zhou, Chun; Liu, Bainan et al. (2014) Transcriptional regulation of ?1H T-type calcium channel under hypoxia. Am J Physiol Cell Physiol 307:C648-56
Rich, Thomas C; Webb, Kristal J; Leavesley, Silas J (2014) Can we decipher the information content contained within cyclic nucleotide signals? J Gen Physiol 143:17-27
Favreau, Peter F; Hernandez, Clarissa; Heaster, Tiffany et al. (2014) Excitation-scanning hyperspectral imaging microscope. J Biomed Opt 19:046010
Villalta, Patricia C; Rocic, Petra; Townsley, Mary I (2014) Role of MMP2 and MMP9 in TRPV4-induced lung injury. Am J Physiol Lung Cell Mol Physiol 307:L652-9
Stevens, Trevor C; Ochoa, Cristhiaan D; Morrow, K Adam et al. (2014) The Pseudomonas aeruginosa exoenzyme Y impairs endothelial cell proliferation and vascular repair following lung injury. Am J Physiol Lung Cell Mol Physiol 306:L915-24
Favreau, Peter; Hernandez, Clarissa; Lindsey, Ashley Stringfellow et al. (2014) Thin-film tunable filters for hyperspectral fluorescence microscopy. J Biomed Opt 19:011017
Xu, Ningyong; Francis, Michael; Cioffi, Donna L et al. (2014) Studies on the resolution of subcellular free calcium concentrations: a technological advance. Focus on "detection of differentially regulated subsarcolemmal calcium signals activated by vasoactive agonists in rat pulmonary artery smooth muscle cells". Am J Physiol Cell Physiol 306:C636-8
Hashizume, Masahiro; Mouner, Marc; Chouteau, Joshua M et al. (2013) Mitochondrial-targeted DNA repair enzyme 8-oxoguanine DNA glycosylase 1 protects against ventilator-induced lung injury in intact mice. Am J Physiol Lung Cell Mol Physiol 304:L287-97
Alexeyev, Mikhail; Shokolenko, Inna; Wilson, Glenn et al. (2013) The maintenance of mitochondrial DNA integrity--critical analysis and update. Cold Spring Harb Perspect Biol 5:a012641
Balczon, Ron; Prasain, Nutan; Ochoa, Cristhiaan et al. (2013) Pseudomonas aeruginosa exotoxin Y-mediated tau hyperphosphorylation impairs microtubule assembly in pulmonary microvascular endothelial cells. PLoS One 8:e74343

Showing the most recent 10 out of 73 publications