The Vascular Biology Core will coordinate a standardized set of measurements across brains and cerebral arteries from all three projects. The main endpoints evaluated by will be smooth muscle phenotype in both large surface arteries and parenchymal vessels imaged in coronal brain slices. Phenotype will be assessed functionally in surface arteries through measurements of myogenic contractility, and structurally in all arteries through measurements of confocal colocalization of contracrile proteins. Each project will prepare their rats, manage the in vivo measurements of behavior and MRI, and then will sacrifice their animals. All brains from these sacrificed animals will be processed in the Vascular Biology Core. Two main categories of analysis will be employed: fresh tissue and fixed tissue. For fresh tissue, surface arteries will be harvested from the brain and used for myography measurements or frozen for immunoblotting measurements of contractile proteins, cell-type specific markers, and other project-specific proteins. For brains fixed in situ, the surface arteries will be removed after which the brains will be embedded, sectioned, and studied using fluorescent immunohistochemistry. The extent and character of changes in smooth muscle phenotype and myogenic function will be compared across arteries obtained from all three hemorrhage models to test our """"""""Gradient Hypothesis"""""""" that all types of cerebral hemorrhage induce cerebrovascular phenotypic transformation whose magnitude varies in relation to distance from the core of injury. In general, one fresh brain will be processed each day, but multiple fixed brains can be processed each day and then analyzed later. For immunohistochemistry and confocal microscopy studies, our current staff processes approximately 20 slides per day, which is more than sufficient to handle the anticipated workload of 6-8 fresh and 18-20 fixed brains each week. Schedules for animal harvesting will be carefully coordinated among the individual projects. This will standardize reagent and resource utilization across all projects, enable volume purchase savings, and eliminate redundancies. All methods to be used in the core have been in routine use for multiple years.

Public Health Relevance

The Vascular Biology Core will employ a variety of immunoblotting, confocal microscopic, and in vitro tissue bath methods to both structurally and functionally assess changes in the expression of brain proteins and cerebrovascular smooth muscle phenotype in response to hemorrhagic injury and treatment in brain samples produced by the three main projects in this PPG.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Program Projects (P01)
Project #
1P01NS082184-01A1
Application #
8661431
Study Section
National Institute of Neurological Disorders and Stroke Initial Review Group (NSD)
Project Start
Project End
Budget Start
2014-01-01
Budget End
2014-12-31
Support Year
1
Fiscal Year
2014
Total Cost
$255,996
Indirect Cost
$93,171
Name
Loma Linda University
Department
Type
DUNS #
009656273
City
Loma Linda
State
CA
Country
United States
Zip Code
92350
Wang, Yuechun; Sherchan, Prativa; Huang, Lei et al. (2017) Naja sputatrix Venom Preconditioning Attenuates Neuroinflammation in a Rat Model of Surgical Brain Injury via PLA2/5-LOX/LTB4 Cascade Activation. Sci Rep 7:5466
Akyol, Gokce Yilmaz; Manaenko, Anatol; Akyol, Onat et al. (2017) IVIG activates Fc?RIIB-SHIP1-PIP3 Pathway to stabilize mast cells and suppress inflammation after ICH in mice. Sci Rep 7:15583
Kim, Cherine H; McBride, Devin W; Sherchan, Prativa et al. (2017) Crotalus helleri venom preconditioning reduces postoperative cerebral edema and improves neurological outcomes after surgical brain injury. Neurobiol Dis 107:66-72
Ma, Li; Manaenko, Anatol; Ou, Yi-Bo et al. (2017) Bosutinib Attenuates Inflammation via Inhibiting Salt-Inducible Kinases in Experimental Model of Intracerebral Hemorrhage on Mice. Stroke 48:3108-3116
Rolland, William B; Krafft, Paul R; Lekic, Tim et al. (2017) Fingolimod confers neuroprotection through activation of Rac1 after experimental germinal matrix hemorrhage in rat pups. J Neurochem 140:776-786
Yu, Lingyan; Lu, Zhengyang; Burchell, Sherrefa et al. (2017) Adropin preserves the blood-brain barrier through a Notch1/Hes1 pathway after intracerebral hemorrhage in mice. J Neurochem 143:750-760
McBride, Devin W; Zhang, John H (2017) Precision Stroke Animal Models: the Permanent MCAO Model Should Be the Primary Model, Not Transient MCAO. Transl Stroke Res :
Hu, Qin; Manaenko, Anatol; Bian, Hetao et al. (2017) Hyperbaric Oxygen Reduces Infarction Volume and Hemorrhagic Transformation Through ATP/NAD+/Sirt1 Pathway in Hyperglycemic Middle Cerebral Artery Occlusion Rats. Stroke 48:1655-1664
Zhang, John H; Obenaus, Andre; Liebeskind, David S et al. (2017) Recanalization, reperfusion, and recirculation in stroke. J Cereb Blood Flow Metab 37:3818-3823
Obenaus, Andre; Ng, Michelle; Orantes, Amanda M et al. (2017) Traumatic brain injury results in acute rarefication of the vascular network. Sci Rep 7:239

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