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Detection and Phenotypic Characterization of Adult Neurogenesis
Authors:H Georg Kuhn  Amelia J Eisch  Kirsty Spalding  Daniel A Peterson
Institution:1.Institute for Neuroscience and Physiology, University of Gothenburg, Gothenburg, SE-405 30, Sweden;2.Department of Psychiatry, University of Texas Southwestern Medical Center, Dallas, Texas 75390-9070;3.Department of Cell and Molecular Biology, Karolinska Institute, Stockholm SE-171 77, Sweden;4.Center for Stem Cell and Regenerative Medicine, Rosalind Franklin University of Medicine and Science, North Chicago, Illinois 60064
Abstract:Studies of adult neurogenesis have greatly expanded in the last decade, largely as a result of improved tools for detecting and quantifying neurogenesis. In this review, we summarize and critically evaluate detection methods for neurogenesis in mammalian and human brain tissue. Besides thymidine analog labeling, cell-cycle markers are discussed, as well as cell stage and lineage commitment markers. Use of these histological tools is critically evaluated in terms of their strengths and limitations, as well as possible artifacts. Finally, we discuss the method of radiocarbon dating for determining cell and tissue turnover in humans.Detection of neurogenesis in vivo requires the ability to image at a cellular resolution, which currently precludes noninvasive imaging approaches, such as magnetic resonance imaging (MRI). In vivo microscopy, using deeply penetrating UV illumination with multiphoton microscopy, or by the recently available endoscopic confocal microscopy, may provide new opportunities for longitudinal studies of neurogenesis in the living animal with single-cell resolution. These newer microscopy approaches are particularly compelling when coupled with transgenic mice expressing phenotype-specific fluorescent reporter genes. Additionally, an advanced method using 14C carbon dating of postmortem DNA from specific cell populations of the brain revealed insights into adult human neurogenesis. Nevertheless, at present, the predominant approach for studying neurogenesis relies on traditional histological methods of fixation, production of tissue sections, staining, and microscopic analysis.This review discusses methodological considerations for detection of neurogenesis in the adult brain according to our current state of knowledge. This will include the use of exogenous or endogenous markers of cell cycle, as well as phenotype markers that contribute to resolving stages of neuronal lineage commitment. The accurate analysis of cell phenotype will be discussed, including suggestions for accurate detection and reliable quantification of cell numbers. Finally, we will present the newly developed 14C carbon dating of nuclear DNA for quantitative analysis of neurogenesis in human tissue.
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