The Sox2 promoter-driven CD63-GFP transgenic rat model allows tracking of neural stem cell-derived extracellular vesicles

Aya Yoshimura; Naoki Adachi; Hitomi Matsuno; Masaki Kawamata; Yusuke Yoshioka; Hisae Kikuchi; Haruki Odaka; Tadahiro Numakawa; Hiroshi Kunugi; Takahiro Ochiya; Yoshitaka Tamai

doi:10.1242/dmm.028779

The Sox2 promoter-driven CD63-GFP transgenic rat model allows tracking of neural stem cell-derived extracellular vesicles

Dis Model Mech. 2018 Jan 30;11(1):dmm028779. doi: 10.1242/dmm.028779.

Authors

Aya Yoshimura^{1

2

3}, Naoki Adachi^{2

4}, Hitomi Matsuno², Masaki Kawamata⁵, Yusuke Yoshioka⁵, Hisae Kikuchi⁶, Haruki Odaka^{2

7

8}, Tadahiro Numakawa^{2

8}, Hiroshi Kunugi², Takahiro Ochiya⁹, Yoshitaka Tamai^{1

10}

Affiliations

¹ Division of Laboratory Animals Resources, National Institute of Neuroscience, National Center of Neurology and Psychiatry (NCNP), 4-1-1 Ogawa-Higashi, Kodaira, Tokyo 187-8502, Japan.
² Department of Mental Disorder Research, National Institute of Neuroscience, NCNP, 4-1-1 Ogawa-Higashi, Kodaira, Tokyo 187-8502, Japan.
³ Education and Research Facility of Animal Models for Human Diseases, Center for Research Promotion and Support, Fujita Health University, 1-98 Dengakugakubo, Kutsukake-cho, Toyoake, Aichi 470-1192, Japan.
⁴ Department of Biomedical Chemistry, School of Science and Technology, Kwansei Gakuin University, 2-1 Gakuen, Sanda, Hyogo 669-1337, Japan.
⁵ Division of Molecular and Cellular Medicine, National Cancer Center Research Institute (NCC), 1-1 Tsukiji 5-chome, Chuo-ku, Tokyo 104-0045, Japan.
⁶ Department of Degenerative Neurological Diseases, National Institute of Neuroscience, NCNP, 4-1-1 Ogawa-Higashi, Kodaira, Tokyo 187-8502, Japan.
⁷ Department of Life Science and Medical Bioscience, School of Advanced Science and Engineering, Waseda University, 3-4-1 Okubo, Shinjuku-ku, Tokyo 169-8555, Japan.
⁸ Department of Cell Modulation, Institute of Molecular Embryology and Genetics, Kumamoto University, 2-2-1 Honjo, Chuo-ku, Kumamoto 860-0811, Japan.
⁹ Division of Molecular and Cellular Medicine, National Cancer Center Research Institute (NCC), 1-1 Tsukiji 5-chome, Chuo-ku, Tokyo 104-0045, Japan tochiya@ncc.go.jp.
¹⁰ Chromocenter Inc., 6-7-4 Minatojima-minami, Chuo-ku, Kobe, Hyogo 650-0047, Japan.

Abstract

Extracellular vesicles (EVs) can modulate microenvironments by transferring biomolecules, including RNAs and proteins derived from releasing cells, to target cells. To understand the molecular mechanisms maintaining the neural stem cell (NSC) niche through EVs, a new transgenic (Tg) rat strain that can release human CD63-GFP-expressing EVs from the NSCs was established. Human CD63-GFP expression was controlled under the rat Sox2 promoter (Sox2/human CD63-GFP), and it was expressed in undifferentiated fetal brains. GFP signals were specifically observed in in vitro cultured NSCs obtained from embryonic brains of the Tg rats. We also demonstrated that embryonic NSC (eNSC)-derived EVs were labelled by human CD63-GFP. Furthermore, when we examined the transfer of EVs, eNSC-derived EVs were found to be incorporated into astrocytes and eNSCs, thus implying an EV-mediated communication between different cell types around NSCs. This new Sox2/human CD63-GFP Tg rat strain should provide resources to analyse the cell-to-cell communication via EVs in NSC microenvironments.

Keywords: CD63; Extracellular vesicles; Model animals; Neural stem cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Animals, Newborn
Astrocytes / metabolism
Brain / growth & development
Brain / metabolism
Cell Differentiation
Coculture Techniques
Embryo, Mammalian / metabolism
Extracellular Vesicles / metabolism*
Green Fluorescent Proteins / metabolism*
Humans
Models, Animal
Neural Stem Cells / metabolism*
Promoter Regions, Genetic*
Rats, Transgenic
SOXB1 Transcription Factors / genetics*
SOXB1 Transcription Factors / metabolism
Spheroids, Cellular / metabolism
Tetraspanin 30 / metabolism*

Substances

SOXB1 Transcription Factors
Tetraspanin 30
Green Fluorescent Proteins