Springer protocols feed by Neuroscience 
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This is an RSS file. You can use it to subscribe to this data in your favourite RSS reader or to display this data on your own website or blog.Analysis of Angiogenesis in the Developing Mouse Central Nervous System
In order to study basic mechanisms of sprouting angiogenesis, researchers worldwide rely on the use of model tissues such as rodent retina, which becomes vascularized postnatally, to study the growth of blood vessels. By definition, models have to be simple, recapitulating angiogenic processes in a stereotyped and relatively easy accessible manner, allowing the application of standardized analyses. These criteria also apply in an ideal manner to the embryonic mouse hindbrain, which becomes vascularized by sprouting angiogenesis from a preformed perineural vascular plexus, leading to the stereotypical formation of a capilla...
Source: Springer protocols feed by Neuroscience - February 12, 2014 Category: Neuroscience Source Type: news
Examining Cerebral Angiogenesis in Response to Physical Exercise
Capillary growth and expansion (angiogenesis) is a prerequisite for many forms of neural and behavioral plasticity. It is commonly observed in both brain and muscle of aerobically exercising animals. As such, several histological methods have been used to quantify capillary density, including perfusion with India ink, various Nissl stains, and immunohistochemistry. In this chapter, we will describe these histological procedures and describe the stereological analysis used to quantify vessel growth in response to aerobic exercise. (Source: Springer protocols feed by Neuroscience)
Source: Springer protocols feed by Neuroscience - February 12, 2014 Category: Neuroscience Source Type: news
Vessel Painting Technique for Visualizing the Cerebral Vascular Architecture of the Mouse
Vessel painting is a simple, cost-effective way to visualize the vascular architecture of the mouse brain and other organs. DiI is a lipophilic carbocyanine dye that binds to lipid membranes and is commonly used for tract tracing in the brain. After perfusion with PBS to remove the blood, perfusion with a special DiI solution allows direct staining of the vasculature. This step is followed by perfusion fixation and removal of the brain from the skull. Pial vessels can be directly imaged using a standard fluorescent microscope. To acquire images of the whole brain, a montage of images at different focal planes is assembled....
Source: Springer protocols feed by Neuroscience - February 12, 2014 Category: Neuroscience Source Type: news
Induction of Cerebral Arteriogenesis in Mice
Unilateral common carotid artery occlusion (CCAO) is a standardized method to initiate collateral artery growth (arteriogenesis) in mouse brain. After CCAO is induced, blood circulation in the circle of Willis is changed and increases shear stress, which triggers increased arterial diameter and improvements in cerebrovascular reserve capacity. Functional improvement can be quantified after experimentally induced stroke by external middle cerebral artery occlusion (MCAO). Stroke volume is evaluated by standard tetrazolium chloride (TTC) staining. Here, we describe in vivo methods of CCAO and MCAO in detail and also the eval...
Source: Springer protocols feed by Neuroscience - February 12, 2014 Category: Neuroscience Source Type: news
A Method of Inducing Global Cerebral Ischemia
The four-vessel occlusion (4-VO) method of global forebrain cerebral ischemia mimics the human clinical condition of cardiac arrest. It results in selective neuronal damage and is a useful experimental system to dissect underlying mechanisms behind ischemic phenomena such as the differential susceptibility of CA1 compared to the CA3 region of the hippocampus. It also provides a “proof-of-principle” system for testing out potential agents for neuroprotection. (Source: Springer protocols feed by Neuroscience)
Source: Springer protocols feed by Neuroscience - February 12, 2014 Category: Neuroscience Source Type: news
Pericytes and Adaptive Angioplasticity: The Role of Tumor Necrosis Factor-Like Weak Inducer of Apoptosis (TWEAK)
The TNF superfamily member TWEAK has emerged as a pleiotropic cytokine that regulates many cellular functions that include immune/inflammatory activity, angiogenesis, cell proliferation, and fate. TWEAK through its inducible receptor, FGF-inducible molecule 14 (Fn14), can induce both beneficial and deleterious activity that has a profound effect on cell survival. Thus it is highly likely that TWEAK and Fn14 expressed by cells of the neurovascular unit help regulate and maintain vascular and tissue homeostasis. In this chapter we discuss the expression of TWEAK and Fn14 signaling in the cerebral microvascular pericyte. Peri...
Source: Springer protocols feed by Neuroscience - February 12, 2014 Category: Neuroscience Source Type: news
Vascular Normalization in Cerebral Angiogenesis: Friend or Foe?
Current antiangiogenic therapies have led to the observation that such agents can lead to improved tumor vessel structure and function termed “vascular normalization” which reduces tumor burden. However, vessel normalization is a transient process, and patients often develop resistance/poor response to anti-vascular strategies that remains an important clinical challenge. Therefore, increasing effort has been made to better understand the cellular and molecular mechanisms of vascular normalization and its contribution to immunomodulation. Herein, we summarize the recent effort to better understand the cellular ...
Source: Springer protocols feed by Neuroscience - February 12, 2014 Category: Neuroscience Source Type: news
Cerebral Angiogenesis: A Realistic Therapy for Ischemic Disease?
Angiogenesis, the sprouting of new capillaries from existing blood vessels, accompanies clinical and experimental stroke and is focused particularly in the salvageable ischemic border zone. As this endogenous angiogenic response correlates positively with clinical prognosis, a more complete understanding of the underlying molecular mechanisms and timing of these events may help in the design of novel therapies for vascular regeneration after stroke. In this review we discuss endogenous protective mechanisms, including angiogenesis and vasculogenesis, and underlying molecular mechanisms. We also consider the feasibility of ...
Source: Springer protocols feed by Neuroscience - February 12, 2014 Category: Neuroscience Source Type: news
Cerebral Angiogenesis During Development: Who Is Conducting the Orchestra?
Blood vessels provide the brain with the oxygen and the nutrients it requires to develop and function. Endothelial cells (ECs) are the principal cell type forming the vascular system and driving its development and remodeling. All vessels are lined by a single EC layer. Larger blood vessels are additionally enveloped by vascular smooth muscle cells (VSMCs) and pericytes, which increase their stability and regulate their perfusion and form the blood–brain barrier (BBB). The development of the vascular system occurs by two processes: (1) vasculogenesis, the de novo assembly of the first blood vessels, and (2) angiogene...
Source: Springer protocols feed by Neuroscience - February 12, 2014 Category: Neuroscience Source Type: news
Scanning Near-Field Optical Microscopy for Investigations of Bio-Matter
Optical near-fields can be employed for a wide range of applications, e.g., light localization, light scattering, and field enhancement. In this chapter the principles of near-field scanning optical microscopy (NSOM) will be outlined. The basic idea of this technique is the extension of the bandwidth of accessible spatial frequencies beyond the limits of conventional light microscopy. This strategy has been implemented in different ways. By now this technique covers a broad spectrum of optical contrasts. Here, special attention is turned on the high-resolution spectroscopic imaging of biological samples. (Source: Springer ...
Source: Springer protocols feed by Neuroscience - February 12, 2014 Category: Neuroscience Source Type: news
Application of Three-Dimensional Structured Illumination Microscopy in Cell Biology: Pitfalls and Practical Considerations
Super-resolution fluorescence microscopy techniques have paved the way to address cell biological questions with unprecedented spatial resolution. Of these, three-dimensional structured illumination microscopy (3D-SIM) reaches a nearly eightfold increased volumetric resolution compared to conventional diffraction-limited methods and allows multicolor optical sectioning of standard fluorescently labeled fixed or live samples. Owing to its broad application spectrum, 3D-SIM is likely to become a key method in cell biological far-field imaging, complementing more specialized higher-resolving techniques, such as single molecul...
Source: Springer protocols feed by Neuroscience - February 12, 2014 Category: Neuroscience Source Type: news
Super-Resolution Fluorescence Microscopy Using Structured Illumination
The resolution of far-field fluorescence microscopy is limited by the Abbe diffraction limit. Making use of the moiré effect, structured illumination microscopy circumvents this limit by projecting fine patterns of light into the sample. From several diffraction limited raw images taken for different pattern positions and orientations, a high resolution image can be calculated. This way, linear structured illumination can enhance the resolution by a factor of about two. Employing nonlinearities such as fluorescence saturation, the resolution can be enhanced even further. In this article, a conceptual as well as a ma...
Source: Springer protocols feed by Neuroscience - February 12, 2014 Category: Neuroscience Source Type: news
Data Analysis for Single-Molecule Localization Microscopy
We review single-molecule localization microscopy techniques with a focus on computational techniques and algorithms necessary for their use. The most common approach to single-molecule localization, Gaussian fitting at positions pre-estimated from local maxima, is illustrated in depth and techniques for two- and three-dimensional data analysis are highlighted. After an introduction explaining the principle requirements of single-molecule localization microscopy, we discuss and contrast novel approaches such as maximum likelihood estimation and model-less fitting. Finally, we give an overview over the existing, scientifica...
Source: Springer protocols feed by Neuroscience - February 12, 2014 Category: Neuroscience Source Type: news
Photoactivated Localization Microscopy for Cellular Imaging
In a method termed photoactivated localization microscopy (PALM), super-resolution fluorescence imaging can be achieved through the localization of single molecules. This allows the resolution of specific proteins fused to the appropriate fluorescent protein label. Here, we summarize fluorescent proteins suitable for PALM, the technical aspects of multicolor and three-dimensional imaging, and the software packages that are available. Additionally, we highlight several biological applications with an emphasis on neuroscience. (Source: Springer protocols feed by Neuroscience)
Source: Springer protocols feed by Neuroscience - February 12, 2014 Category: Neuroscience Source Type: news
Application of Real-Time STED Imaging to Synaptic Vesicle Motion
In the emerging field of super-resolution microscopy, the branch of live-cell imaging is still in its infancy. Regardless of its importance for addressing relevant biological questions, live super-resolution imaging has to face several obstacles when compared to conventional imaging: (1) speeding up the naturally slow image acquisition process, (2) choosing appropriate fluorophores (both in terms of photostability and spectral properties), and (3) handling increased illumination intensities (as usually higher intensities are needed for live imaging at adequate frequencies compared to fixed-cell imaging). In this chapter, w...
Source: Springer protocols feed by Neuroscience - February 12, 2014 Category: Neuroscience Source Type: news
