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|  Oregon Health and Science University |
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| Oregon Health and Science University |
My lab is interested in understanding the changes that occur in the cardiac innervation following ischemia reperfusion (a heart attack). Alterations in the sympathetic innervation of the heart after myocardial infarction can trigger arrhythmias and sudden cardiac death. These neuronal changes are not well understood, but blocking cardiac receptors for the neurotransmitter norepinephrine (NE) can help prevent arrhythmias. We are trying to understand the molecular basis for these changes in noradrenergic function, investigating the regulation of neuronal proteins that synthesize, store, and remove NE, and the genes that encode them. We are also examining the induction of neuropeptides in the cardiac innervation following infarction, and how those peptides alter neurotransmission and cardiac function. Finally, we are examining the role of neurotrophins in post-infarct denervation and nerve sprouting. Cardiac nerve sprouting in humans has been directly linked to cardiac pathology and sudden cardiac death. We use a variety of molecular, biochemical, and histological techniques to investigate the regulation of these proteins and genes, using cell lines, primary neuronal cultures, and whole animal studies.
"DSI implantable telemetry transmitters have been, and continue to be, used in studies involving mice such as telemetric recordings of sleep and home cage activity, abnormal heart rate and body temperature in mice lacking thyroid hormone receptor x1, the effects of triiodothyronine on heart rate, and numerous other studies."
Dual pump system for baseline flattening.
Millar SPR1000 for mice. Tip size is 1F.
1.0 F size (for small animals).
Includes heat pad, ventilator for anethesia and oxygen, dissecting microscope.
This is an easy-to-use upright microscope designed specifically for slide viewing.
gp130 is the cytokine receptor subunit that is essential for the action of all IL-6 cytokine family members.
Need MTA from Hermann Rohrer (Max Plank Frankfurt).
"B6.129S4- Ngfrtm1Jae/J mice contain two mutated exon III alleles, which removes the coding region for the last three of four cysteine-rich repeats in the extracellular domain and prevents the expression of functional p75NTR."
From Jackson Labs.
Need MTA from collaborator from McGill, Dr. Michael Tremblay.
DBH expressing neurons lack Stat3.
Need MTA from Hermann Rohrer (Max Plank Frankfurt).
Used to study sympathetic innervation of the heart and axon outgrowth.
Note: all protocols are subject to availability of personnel for training.
Immunohistochemistry protocol for mouse heart tissue with antibodies listed below.
Note: all protocols are subject to availability of personnel for training.
Protocol measures uptake of 3H-NA into sympathetic nerve terminals.
Note: all protocols are subject to availability of personnel for training.
Sympathetic neurons are dissociated from the superior cervical ganglia of newborn rat pups.
Note: all protocols are subject to availability of personnel for training.
Need permission from Dona Chikaraishi at Duke University.
"Tyrosine hydroxylase plays an important role in the physiology of adrenergic neurons. It is the first and rate-limiting enzyme involved in the biosynthesis of the catecholamines Dopamine and Norepinephrine from tyrosine. TH is, therefore, a useful marker for dopaminergic and noradrenergic neurons. The enzymatic activity of TH requires ferrous ions as cofactors and is believed to be regulated by phosphorylation. At least four isoforms of human TH have been identified which result from alternative splicing."
"Tyrosine hydroxylase plays an important role in the physiology of adrenergic neurons. It is the first and rate-limiting enzyme involved in the biosynthesis of the catecholamines Dopamine and Norepinephrine from tyrosine. TH is, therefore, a useful marker for dopaminergic and noradrenergic neurons. The enzymatic activity of TH requires ferrous ions as cofactors and is believed to be regulated by phosphorylation. At least four isoforms of human TH have been identified which result from alternative splicing."
Antibody was gift from Dr. Story Landis (Tyrrell and Landis, 1994).
The Arix/Phox2a expression construct was a generous gift from Dr. Elaine Lewis (NIH).
Used as analgesic for post-op rodents.
"Brain (CATH.a)... cell lines have been established that synthesize abundant dopamine and norepinephrine and express the appropriate catecholaminergic biosynthetic enzymes, tyrosine hydroxylase (TH) and dopamine beta-hydroxylase. The line [was] derived from TH-positive tumors in transgenic mice carrying the SV40 T antigen oncogene under the transcriptional control of 773 base pairs of 5' flanking sequences from the rat TH gene. Although the lines continue to express T antigen, they exhibit neuronal properties such as neurofilaments and synaptophysin and lack glial intermediate filaments. Although in vivo TH is only expressed in postmitotic neurons in the CNS, the CATH.a line demonstrates that TH expression and continued cell division are not incompatible after oncogenic transformation."
The Hand2 expression construct was a generous gift from Dr. Peter Cserjesi (Tulane University, New Orleans, LA). Would need permission from Peter Cserjesi to use this construct.
"Adrenal (PATH.1) cell lines have been established that synthesize abundant dopamine and norepinephrine and express the appropriate catecholaminergic biosynthetic enzymes, tyrosine hydroxylase (TH) and dopamine beta-hydroxylase. The line [was] derived from TH-positive tumors in transgenic mice carrying the SV40 T antigen oncogene under the transcriptional control of 773 base pairs of 5' flanking sequences from the rat TH gene. Although the lines continue to express T antigen, they exhibit neuronal properties such as neurofilaments and synaptophysin and lack glial intermediate filaments."
Pheochromacytoma cell line.
From Kwang Su Kim's laboratory. Would need permission.
Human neuroblastoma cell line.
Chart Pro provides users using PowerLab data acquisition systems with acquisition and analysis capabilities.
This software collects and processes data from telemetry systems. "One of DSI acquisition and analysis system’s that allows researchers to monitor and collect physiologic data from conscious, freely moving laboratory animals. Physiological parameters that are monitored by DSI Transmitters and transmitted via radio frequency signals to a variety of DSI receivers can be collected and analyzed by Dataquest A.R.T.
DSI’s Dataquest A.R.T.™ system collects digital data with accuracy, reliability and ease, and maintains digital data from the receiver to the computer to optimize data integrity throughout the system. The entire family of PC-based Dataquest A.R.T.™ software, including Dataquest® OpenART and A.R.T. Analysis is ideal for collecting telemetry information, no matter the number or size of animals being studied. Sampling rate, calibrations, implant type, animals monitored, extracted features, and other protocol settings can be easily selected using a Windows® graphical user interface."
"GraphPad Prism is a powerful combination of basic biostatistics, curve fitting and scientific graphing in one comprehensive program....
Designed for the practical scientist, Prism does not expect you to be a statistician. It guides you through each analysis - giving you as much help as you need - and tracks and organizes your work like no other program available."
"ImageJ is a public domain, Java-based image processing program developed at the National Institutes of Health. ImageJ was designed with an open archit"ImageJ is a public domain Java image processing program inspired by NIH Image for the Macintosh. It runs, either as an online applet or as a downloadable application, on any computer with a Java 1.4 or later virtual machine. Downloadable distributions are available for Windows, Mac OS, Mac OS X and Linux.
It can display, edit, analyze, process, save and print 8-bit, 16-bit and 32-bit images. It can read many image formats including TIFF, GIF, JPEG, BMP, DICOM, FITS and "raw". It supports "stacks", a series of images that share a single window. It is multithreaded, so time-consuming operations such as image file reading can be performed in parallel with other operations.
It can calculate area and pixel value statistics of user-defined selections. It can measure distances and angles. It can create density histograms and line profile plots. It supports standard image processing functions such as contrast manipulation, sharpening, smoothing, edge detection and median filtering."
"NIS-Elements AR provides a total imaging solution covering image capture, archiving, and analysis. It was developed by Nikon for the most advanced levels of research—giving the user complete control of all microscope and camera functions.
The software is geared to deliver power and maximize workflow. It handles multi-dimensional imaging flawlessly, with support for capture, display, peripheral device control, and data management and analysis of up to six dimensions (X,Y,Z,Lambda(wavelength),T, multipoint). It also offers sophisticated image processing features, such as an extremely powerful deconvolution module, exclusive one-click database capability, and Extended Depth of Focus function.
The system contributes to experiment efficiency with a database building feature developed to handle archiving and searching of large numbers of multi-dimensional image files.
Unified control of the entire imaging system offers significant benefits to users for cutting-edge research, such as live cell imaging."
LabWorks is image analysis software.
