Single-Cell Analysis for Glycogen Localization and Metabolism in Cultured Astrocytes
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ORIGINAL RESEARCH
Single‑Cell Analysis for Glycogen Localization and Metabolism in Cultured Astrocytes Yuanyuan Zhu1 · Ze Fan2 · Rui Wang1 · Rougang Xie1 · Haiyun Guo2 · Ming Zhang1 · Baolin Guo1 · Tangna Sun1 · Haifeng Zhang1 · Lixia Zhuo3 · Yan Li3 · Shengxi Wu1 Received: 29 August 2019 / Accepted: 8 December 2019 © The Author(s) 2019
Abstract Cerebral glycogen is principally localized in astrocytes rather than in neurons. Glycogen metabolism has been implicated in higher brain functions, including learning and memory, yet the distribution patterns of glycogen in different types of astrocytes have not been fully described. Here, we applied a method based on the incorporation of 2-NBDG, a d-glucose fluorescent derivative that can trace glycogen, to investigate glycogen’s distribution in the brain. We identified two types of astrocytes, namely, 2-NBDGI (glycogen-deficient) and 2-NBDGII (glycogen-rich) cells. Whole-cell patch-clamp and fluorescence-activated cell sorting (FACS) were used to separate 2-NBDGII astrocytes from 2-NBDGI astrocytes. The expression levels of glycogen metabolic enzymes were analyzed in 2-NBDGI and 2-NBDGII astrocytes. We found unique glycogen metabolic patterns between 2-NBDGI and 2-NBDGII astrocytes. We also observed that 2-NBDGII astrocytes were mainly identified as fibrous astrocytes but not protoplasmic astrocytes. Our data reveal cell type-dependent glycogen distribution and metabolism patterns, suggesting diverse functions of these different astrocytes. Keywords Astrocytes · Glycogen · Glycogen metabolism · Single-cell PCR · Fibrous astrocyte · Protoplasmic astrocyte
Introduction Glial cells are the main type of neural cell and exist throughout the central nervous system (CNS) (Gallo and Deneen 2014; Brosius Lutz and Barres 2014; Walsh et al. 2014). Estimates regarding the ratio of glial cells to neurons vary Yuanyuan Zhu, Ze Fan and Rui Wang are Co-first authors. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10571-019-00775-4) contains supplementary material, which is available to authorized users. * Yan Li [email protected] * Shengxi Wu [email protected] 1
Department of Neurobiology, The School of Basic Medicine, The Fourth Military Medical University, Xi’an, China
2
Department of Anesthesiology and Perioperative Medicine, Xijing Hospital of the Fourth Military Medical University, Xi’an, China
3
Center for Brain Science, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China
greatly. However, the number of glial cells likely is at least equal to or exceeds the number of neurons. Among glial cells in mammalian brains, 20–40% are specifically defined as astrocytes, although the percentage of astrocytes has considerable variability across species and brain areas (Khakh and Sofroniew 2015). Astrocytes play important roles in the CNS, including roles in brain development, synaptic plasticity, synaptic transmission, blood flow regulation, energy metabolism, blood–brain barrier formation, cir
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