Visualization of trans homophilic interaction of clustered protocadherin in neurons

Journal Proc. Natl. Acad. Sci. U. S. A. 120(38):e2301003120. (2023)
Title Visualization of trans homophilic interaction of clustered protocadherin in neurons
Laboratory KOKORO-Biology Group〈Prof. YAGI Takeshi〉

Clustered protocadherin (Pcdh) functions as a cell recognition molecule through the homophilic interaction in the central nervous system. However, its interactions have not yet been visualized in neurons. We previously reported PcdhγB2-Förster resonance energy transfer (FRET) probes to be applicable only to cell lines. Herein, we designed γB2-FRET probes by fusing FRET donor and acceptor fluorescent proteins to a single γB2 molecule and succeeded in visualizing γB2 homophilic interaction in cultured hippocampal neurons. The γB2-FRET probe localized in the soma and neurites, and FRET signals, which were observed at contact sites between neurites, eliminated by ethylene glycol tetraacetic acid (EGTA) addition. Live imaging revealed that the FRET-negative γB2 signals rapidly moved along neurites and soma, whereas the FRET-positive signals remained in place. We observed that the γB2 proteins at synapses rarely interact homophilically. The γB2-FRET probe might allow us to elucidate the function of the homophilic interaction and the cell recognition mechanism.


Natsumi Hoshino (1), Takashi Kanadome (2, 3), Tomomi Takasugi (1), Mizuho Itoh (1), Ryosuke Kaneko (1), Yukiko U Inoue (4), Takayoshi Inoue (4), Takahiro Hirabayashi (1, 5), Masahiko Watanabe (6), Tomoki Matsuda (2), Takeharu Nagai (2), Etsuko Tarusawa (1), Takeshi Yagi (1)

  1. KOKORO-Biology Group, Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan.
  2. Department of Biomolecular Science and Engineering, SANKEN, Osaka University, Ibaraki, Osaka 567-0047, Japan.
  3. Japan Science and Technology Agency, Precursory Research for Embryonic Science and Technology, Kawaguchi, Saitama 332-0012, Japan.
  4. Department of Biochemistry and Cellular Biology, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Tokyo 187-8501, Japan.
  5. Clinical Medicine Research Laboratory, Shonan University of Medical Sciences, Yokohama 244-0806, Japan.
  6. Department of Anatomy, Faculty of Medicine, Hokkaido University, Sapporo, Hokkaido 060-8638, Japan.
PubMed 37695902