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Evaluation of Rho-Kinase Activity in Mice Brain Using N-[11C]Methyl-hydroxyfasudil with Positron Emission Tomography

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Abstract

Purpose

Rho is a small molecular weight GTP-binding protein and works as a molecular shuttling switch between an active (GTP-bound) and inactive (GDP-bound) state. Rho is known to be involved in cell motility, cell adhesion, and cytokinesis through actin cytoskeleton reorganization. The GTP-bound form of Rho interacts with its specific downstream target, triggering intracellular signaling cascades. Rho effectors such as Rho-kinases have been isolated on the basis of their selective binding to the GTP-bound form of Rho. Rho-kinase is thought to have an important role in the pathogenesis of a variety of neurological diseases because activation of the Rho/Rho-kinase pathway has been observed in various central nervous system disorders. Previous histochemical studies have shown multiple molecular mechanisms for the regulation of Rho-kinase. Neuroimaging of Rho/Rho-kinase has rarely been studied because of a lack of appropriate radiotracers. Recently, N-[11C]methyl-hydroxyfasudil, a new radiotracer for positron emission tomography (PET), has been introduced to measure Rho-kinase activity. In this study, the regional distribution and kinetics of N-[11C]methyl-hydroxyfasudil were investigated in the brains of mice.

Procedures

A 90-min dynamic scan was performed following intravenous infusion of N-[11C]methyl-hydroxyfasudil.

Results

The uptake of N-[11C]methyl-hydroxyfasudil reached a maximum within 5 min and gradually decreased in all organs. The standard uptake values (SUVs) in the brain, liver, and kidney on average between 30 to 60 min were 0.17 ± 0.03, 0.76 ± 0.18, and 0.62 ± 0.18 and from 60 to 90 min were 0.15 ± 0.01, 0.69 ± 0.33, and 0.64 ± 0.17, respectively. N-[11C]Methyl-hydroxyfasudil showed a widespread distribution throughout the brain, with low levels of radioactivity. Radioactivity concentration in plasma at 90 min after injection of N-[11C]methyl-hydroxyfasudil resulted in SUVs in the control and fasudil pretreatment of 0.0013 and 0.0023 ± 0.0008, respectively. Compared to normal control mice, about twofold higher radioactivity concentration was observed in fasudil-pretreated mice. In a cold brain injury mouse model, accumulation of N-[11C]methyl-hydroxyfasudil was slightly higher at the injury site than that at the control site, and the difference was statistically significant in the “24 h after injury” group (P < 0.05).

Conclusions

These results suggest that following brain injury, N-[11C]methyl-hydroxyfasudil binds to the active form of Rho-kinase. PET imaging using N-[11C]methyl-hydroxyfasudil could provide new insights into the pathophysiology of a variety of neurological disorders including stroke, inflammatory diseases, demyelinating diseases, Alzheimer's disease, and neuropathic pain.

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Acknowledgments

We thank Mr. Hidekatsu Wakizaka of the National Institute of Radiological Sciences for his assistance with the PET experiments.

Conflicts of interest

The authors declare no conflict of interest.

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Authors and Affiliations

  1. Molecular Imaging Center, National Institute of Radiological Sciences, 4-9-1 Anagawa, Inage-ku, Chiba, 263-8555, Japan

    Junko Taniguchi, Chie Seki, Hiroyuki Takuwa, Hiroshi Kawaguchi, Yoko Ikoma, Masayuki Fujinaga, Iwao Kanno, Ming-Rong Zhang & Hiroshi Ito

  2. Department of Neurology, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8670, Japan

    Junko Taniguchi & Satoshi Kuwabara

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  1. Junko Taniguchi
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  2. Chie Seki
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Correspondence to Hiroshi Ito.

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Taniguchi, J., Seki, C., Takuwa, H. et al. Evaluation of Rho-Kinase Activity in Mice Brain Using N-[11C]Methyl-hydroxyfasudil with Positron Emission Tomography. Mol Imaging Biol 16, 395–402 (2014). https://doi.org/10.1007/s11307-013-0695-y

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  • DOI: https://doi.org/10.1007/s11307-013-0695-y

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