AI Article Synopsis
- - The study focuses on using inducible transgenic Nestin-CreER mice to research gene targeting and molecular pathways related to adult neurogenesis, specifically in areas of the brain like the SGZ and SVZ.
- - Researchers evaluated the effectiveness of genetic recombination in the aging SVZ by comparing 12-month-old and 20-month-old mice to 2-month-old mice using a specific mouse line and various molecular techniques.
- - The analysis involved measuring Cre mRNA and YFP gene expression, along with co-immunolabeling, to assess the percentage of progeny that were successfully targeted and the cell proliferation rates in aging mice.
Article Abstract
The use of inducible transgenic Nestin-CreER mice has proved to be an essential research tool for gene targeting and studying the molecular pathways implicated in adult neurogenesis, namely, inside the adult subgranular zone (SGZ) of the dentate gyrus and the adult subventricular zone (SVZ) lining the lateral ventricles. Several lines of Nestin-CreER-expressing mice were generated and used in adult neurogenesis research in the past two decades; however, their suitability for studying neurogenesis in aged mice remains elusive. Here, we assessed the efficiency of Cre-loxP genetic recombination in the aging SVZ using the Nestin-CreER/Rosa26YFP line designed by Lagace et al. (J Neurosci 27(46):12623-12629, 2007). This analysis was performed in 12-month-old (middle-aged) mice and 20-month-old (old) mice compared to 2-month-old (young adult) mice. To evaluate successful recombination, our approach relies on the histological assessment of Cre mRNA level of expression and the YFP reporter gene's expression inside the aging SVZ by combining in situ hybridization and immunohistochemistry. Using co-immunolabeling, this approach also provides the advantage of estimating the percentage of recombined progeny [(GFP+Nestin+)/Nestin+] and the rate of cell proliferation [(GFP+Ki67+)/GFP+] inside the aging SVZ niche.
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| http://dx.doi.org/10.1007/7651_2019_214 | DOI Listing |
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