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Alteration of the neuronal and glial cell profiles in Neu1-deficient zebrafish

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Abstract

Neu1 is a glycosidase that releases sialic acids from the non-reducing ends of glycoconjugates, and its enzymatic properties are conserved among vertebrates. Recently, Neu1-KO zebrafish were generated using genome editing technology, and the KO fish showed abnormal emotional behavior, such as low schooling, low aggressiveness, and excess exploratory behavior, accompanied by the downregulation of anxiety-related genes. To examine the alteration of neuronal and glial cells in Neu1-KO zebrafish, we analyzed the molecular profiles in the zebrafish brain, focusing on the midbrain and telencephalon. Using immunohistochemistry, we found that signals of Maackia amurensis (MAM) lectin that recognizes Sia α2-3 linked glycoconjugates were highly increased in Neu1-KO zebrafish brains, accompanied by an increase in Lamp1a. Neu1-KO zebrafish suppressed the gene expression of AMPA-type glutamate receptors such as gria1a, gria2a, and gria3b, and vesicular glutamate transporter 1. Additionally, Neu1-KO zebrafish induced the hyperactivation of astrocytes accompanied by an increase in Gfap and phosphorylated ERK levels, while the mRNA levels of astrocyte glutamate transporters (eaat1a, eaat1c, and eaat2) were downregulated. The mRNA levels of sypb and ho1b, which are markers of synaptic plasticity, were also suppressed by Neu1 deficiency. Abnormal activity of microglia was also revealed by IHC, and the expressions of iNOS and IL-1β, an inflammatory cytokine, were increased in Neu1-KO zebrafish. Furthermore, drastic neuronal degeneration was detected in Neu1-KO zebrafish using Fluoro-Jade B staining. Collectively, the neuronal and glial abnormalities in Neu1-KO zebrafish may be caused by changes in the excitatory neurotransmitter glutamate and involved in the emotional abnormalities.

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Acknowledgements

We appreciate the support of Dr. Yumiko Yoshizaki, Dr. Shinji Kanda, Dr. Takanori Ikenaga, and the technical assistance of Mika Ishii, Momoko Kawabe, Chihoko Horita, Takumi Nishida, Kanta Miyamoto, Toshiki Hyodo, and Shoji Kodama. The financial support from Sankei Science Foundation is gratefully acknowledged.

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  1. Course of Biological Science and Technology, The United Graduate School of Agricultural Sciences, Kagoshima University, 1-21-24 Korimoto, Kagoshima, 890-0065, Japan

    Asami Ikeda, Masaharu Komatsu & Kazuhiro Shiozaki

  2. Department of Food Life Sciences, Faculty of Fisheries, Kagoshima University, 4-50-20 Shimo-arata, Kagoshima, 890-0056, Japan

    Chiharu Yamasaki, Yurina Kubo, Yudai Doi, Mayu Komamizu, Masaharu Komatsu & Kazuhiro Shiozaki

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  1. Asami Ikeda
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  2. Chiharu Yamasaki
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Contributions

Conceptualization: Asami Ikeda, and Kazuhiro Shiozaki; Methodology: Asami Ikeda, Chiharu Yamasaki, and Kazuhiro Shiozaki; Formal analysis and investigation: Asami Ikeda, Chiharu Yamasaki, Yurina Kubo, Yudai Doi, Mayu Komamizu, and Kazuhiro Shiozaki; Writing -original draft preparation: Asami Ikeda and Kazuhiro Shiozaki; Writing review and editing: Masaharu Komatsu and Kazuhiro Shiozaki; Supervision: Masaharu Komatsu and Kazuhiro Shiozaki.

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Correspondence to Kazuhiro Shiozaki.

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Ikeda, A., Yamasaki, C., Kubo, Y. et al. Alteration of the neuronal and glial cell profiles in Neu1-deficient zebrafish. Glycoconj J 39, 499–512 (2022). https://doi.org/10.1007/s10719-022-10074-8

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