Spatial and Intracellular Distribution of the Endogenous Calcineurin-Inhibitory Proteins, ZAKI-4, in Mouse Brain

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  • Hoshino Shin
    Division of Molecular and Cellular Adaptation, Research Institute of Environmental Medicine, Nagoya University
  • Takagishi Yoshiko
    Division of Molecular and Cellular Adaptation, Research Institute of Environmental Medicine, Nagoya University
  • Kanou Yasuhiko
    Division of Molecular and Cellular Adaptation, Research Institute of Environmental Medicine, Nagoya University
  • Hayasaka Shizu
    Division of Molecular and Cellular Adaptation, Research Institute of Environmental Medicine, Nagoya University
  • Hattori Kimihiko
    Division of Molecular and Cellular Adaptation, Research Institute of Environmental Medicine, Nagoya University
  • Kambe Fukushi
    Division of Molecular and Cellular Adaptation, Research Institute of Environmental Medicine, Nagoya University
  • Seo Hisao
    Division of Molecular and Cellular Adaptation, Research Institute of Environmental Medicine, Nagoya University
  • Murata Yoshiharu
    Division of Molecular and Cellular Adaptation, Research Institute of Environmental Medicine, Nagoya University

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Abstract

ZAKI-4 is a thyroid hormone-responsive gene which encodes two isoforms, ZAKI-4 α and β, both of which belong to a family of proteins that inhibit calcineurin activity. Calcineurin is a calcium/calmodulin-dependent phosphatase and is known to play crucial roles in brain development and function. Using in situ hybridization and immunohistochemistry, the present study aimed to demonstrate regional distribution of ZAKI-4 mRNAs and proteins in the mouse brain. Both ZAKI-4 mRNA isoforms showed similar spatial expression in the brain. Although they were widely expressed throughout the entire brain, the highest expression was observed in neurons in olfactory bulb, hippocampus and cerebellum. Consistent with the expression of ZAKI-4 mRNA isoforms, ZAKI-4 proteins were widely, but not evenly, distributed in the brain; the most intense immunoreactivity was found in the olfactory bulb, cerebral neocortex, hippocampus and cerebellum. As for the subcellular localization, ZAKI-4 immunoreactivity was confined to neuronal somata, with higher expression in the soma than in dendrites, and was not detected in glia. The double immunostaining of ZAKI-4 proteins and calcineurin revealed that they were co-localized in the periphery of the soma and dendrites of neurons. These results indicate that ZAKI-4 proteins are localized in the area where they are able to inhibit calcineurin activity.<br>

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