Changing stroke rehab and research worldwide now.Time is Brain! trillions and trillions of neurons that DIE each day because there are NO effective hyperacute therapies besides tPA(only 12% effective). I have 523 posts on hyperacute therapy, enough for researchers to spend decades proving them out. These are my personal ideas and blog on stroke rehabilitation and stroke research. Do not attempt any of these without checking with your medical provider. Unless you join me in agitating, when you need these therapies they won't be there.

What this blog is for:

My blog is not to help survivors recover, it is to have the 10 million yearly stroke survivors light fires underneath their doctors, stroke hospitals and stroke researchers to get stroke solved. 100% recovery. The stroke medical world is completely failing at that goal, they don't even have it as a goal. Shortly after getting out of the hospital and getting NO information on the process or protocols of stroke rehabilitation and recovery I started searching on the internet and found that no other survivor received useful information. This is an attempt to cover all stroke rehabilitation information that should be readily available to survivors so they can talk with informed knowledge to their medical staff. It lays out what needs to be done to get stroke survivors closer to 100% recovery. It's quite disgusting that this information is not available from every stroke association and doctors group.

Monday, March 19, 2012

Cellular Imaging and Emerging Technologies for Adult Neurogenesis Research

A good overview of neurogeneisis, confound your doctor by asking specific questions about it.
http://www.frontiersin.org/Journal/FullText.aspx?ART_DOI=10.3389/fnins.2012.00041&name=Neurogenesis
The first report on the generation of new neurons in the adult mammalian brain occurred in the early ‘60s, however, nearly 40 years passed before the scientific community generally recognized the existance of adult mammalian neurogenesis. Development of new technologies that facilitate the identification of new-born neurons in the early ‘90s has been central to expanding our understanding of adult neurogenesis as a process influencing mammalian brain plasticity. Subsequently, the field of adult neurogenesis progressed tremendously thanks to continuous technical advances allowing in vivo and in vitro manipulation of adult neuronal progenitors. Today, a core understanding of various aspects of adult neurogenesis has emerged, including neural progenitor proliferation and fate-specification, and the migration, maturation and synaptic integration of new-born neurons into functional circuits, but numerous open questions remain. This research topic issue gathers a series of articles dedicated to major methodological advancements that in the recent years have significantly contributed to progress in the understanding of adult neurogenesis in the mammalian brain. It includes six review articles that give a critical update of current approaches, outlining their strengths as well as their limits, and the need of further improvement in technological tools to address specific key issues. Four methods articles dealing with new in vitro and in vivo or ex vivo approaches to study adult neurogenesis, together with one original research article are included.
In vitro assays are key tools for deciphering the cellular and molecular mechanisms of adult neurogenesis. Generally, adult neural precursor cells can be expanded in vitro using two different culture methodologies: neurospheres, or adherent monolayer cultures. Babu et al., present a detailed protocol for isolation and enrichment of neural precursor cells from mouse adult hippocampal neurons and highlight potential modifications to the protocol useful for isolating and expanding precursor cells from other brain regions.
Newly generated neurons in vivo can be identified and/or manipulated by various approaches, such as the incorporation of nucleotide analogues, retrovirus-mediated gene transfer, and genetic methods using transgenic mice. Among nucleotide analogue methods, BrdU has been the marker of choice in recent years. BrdU labelling of dividing cells allows various analysis including birth-dating, cell cycle analysis and evaluation of selective survival of newly generated cells following different experimental paradigms. Sultan et al., present original data obtained by coupling BrdU and activated caspase-3 labelling to cellular mapping. They analysed the number, spatial distribution, and apoptosis of newly born cells in the granule cell layer of the olfactory bulb following olfactory conditioning paradigms. This revealed a region-specific reduction in new-born cell death that correlates with the time point at which animals have acquired the task. However, a key limitation of the BrdU method is its ability to recognize only a single pool of BrdU positive cells at one time in the same animal. llorens-Martin and Trejo describe the recent development of multiple birth-dating analysis involving the injection of different thymidine analogues (i.e IdU and CldU), that can be unequivocally distinguished using specific antibodies, allowing two to three cell subpopulations of different ages to be labelled in the same animal. Focusing on the hippocampus, the authors describe the main results obtained by this technique, outlining some of the key applications as well as the main concerns associated with multi-dating approaches.
Recent developments in genetic methods to identify and manipulate newly born neurons are invaluable tools to progress in the study of adult neurogenesis. Imayoshi et al. specifically discuss this topic describing the application of site-specific recombinases and Tet inducible system in combination with transgenic or gene targeting strategy. They present several genetic models to suppress adult neurogenesis. An alternative method to deplete adult generated neurons is described in the article by Tan et al. that give technical details and results obtained by image-guided irradiation at different doses and survival time. Among the additional manipulation possibilities Imayoshi et al. also discuss the need to develop new genetic techniques to effectively increase neurogenesis in brain regions that are normally non neurogenic. In a further refinement of genetic approaches, the review by Benjamin Arenkiel describes approaches to reveal connectivity of newly born neurons in the adult brain, and to manipulate cell and circuit activity. He summarizes the current viral tracing methods, heterologous receptor expression systems, and optogenetic technologies that hold promise toward elucidating the wiring and circuit properties of adult-born neurons.
Two methods papers in this special issue detail new approaches to analyse integration of adult born neurons relaying on immunohistochemical detection of endogenous markers. Rosenzweig and Wojtowicz describe the development of a method for ex-vivo analysis of dendritic growth in immature adult-born hippocampal granule neurons. The method is based on laminar quantification of cell bodies, primary, secondary and tertiary dendrites separately and independently from each other. The data demonstrate the suitability of this technique for analysis of dendritic growth and complexity comparing different experimental conditions. Luzzati et al., developed a new procedure suitable to analyse neurogenesis in parenchymal neurogenic niches, which represents a reliable alternative to the whole mount approaches to analyse cyto-architectural features of adult germinative niche. The method, based on existing freeware software, combine confocal laser scanning microscopy and serial section reconstruction in order to span large volumes of brain tissue at cellular resolution. An example is described by investigating the morphology and spatial organization of a group of doublecortin positive neuroblasts located in the lateral striatum of the late post-natal guinea pig.
The latest progress towards in vivo imaging of neurogenesis in animal models are discussed in the reviews by James et al. and Couillard-Despres et al. The first review centers on quantitative multi-photon microscopy applied to the dynamic study of SVZ-derived neuroblast migration in acute slices. In this paper James et al., present a brief overview of SVZ-neuroblast time-lapse imaging studies and the current knowledge of cellular patterns of SVZ neuroblast migration, highlighting putative underlying regulatory mechanisms. They give technical suggestions useful when setting up a two-photon microscope imaging system and identify several unsolved questions about SVZ neuroblast migration that might be addressed with current or emerging strategies to further harness the deep potential of two-photon microscopy. Couillard-Despres et al. present a comprehensive review on in vivo monitoring of adult neurogenesis describing the use of magnetic resonance imaging (MRI), positron emission tomography (PET) and optical imaging with fluorescent or bioluminescent reporters. They compare the different approaches and outline one major challenge in the achievement of neurogenesis-specificity and detection-sensitivity for direct (e.g., iron oxide particles, [18F]-FLT) and indirect (i.e., the use of reporter-genes) labelling respectively. The authors observe that, although preclinical settings in animal models offer the possibility to use transgenic reporter systems, the development of these imaging techniques to the study of adult neurogenesis in humans remains the ultimate goal. That issue is further highlighted in the paper by Sierra et al., which focus on the human brain detailing the first data on adult human neurogenesis starting from methodologies directly imported from the rodent research up to humans. The authors describe new technologies specifically developed for the detection and quantification of neural stem cells in the living human brain. These technologies rely on the use of MRI, available in hospitals worldwide and hold the potential to test the contribution of adult human neurogenesis to brain function in both health and disease.

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