National Institute for Basic Biology

Developmental Biology

LABORATORY OF REGENERATION BIOLOGY
Director General

AGATA, Kiyokazu

Specially Appointed Associate Professor:
SUZUKI, Ken-ichi
Postdoctoral Fellow:
TERAMOTO, Machiko
SOKENDAI Graduate Student: SUGIURA, Nao
KUROKI, Yoshihito
BO, Kazuto
Visiting Graduate Student: ISHIDA, Miyuki
Technical Assistant:
KAJIURA-KOBAYASHI, Hiroko
Admin Support Staff:
SAKAGAMI, MariNISHIMURA, NorikoComparative Regenerative Biology

We use animals that demonstrate a high ability in regenerating body parts, such as planarians and newts, to understandthe principle of regeneration. In particular, we investigate
the difference between regenerative and non-regenerative
animals to evoke abilities from non-regenerative animals.
We have already succeeded in achieving this with planarians,
which were able to regenerate their heads through RNAi
(Umesono et al., 2013 Nature), and accomplishing functional
joint regeneration in frogs through the activation of reintegration systems (Tsutsumi et al., 2016 Regeneration). We
are currently trying to induce functional joint regenerationin mouse and to characterize adult pluripotent stem cells inplanarian at a single cell level.

staining with several fluorescence dyes. Recently, we found acondition to be able to isolate viable aPSC by FACS sorting
without staining with dyes. The single cell transcriptome
analyses using these isolated aPSC showed unique propertiesof planarian aPSC (Kuroki et al., unpublished).
We also found that these aPSC showed the collective migration property (branch-like patterns in Fig.1) after RNAitreatment of the MTA (Metastatic Tumor Antigen) family
genes, suggesting that planarian might have unique niche formaintain aPSC in the adult bodies (Sato et al., 2022).

Figure.1 RNAi treated animal of the MTA-A gene showed the branch-like
distribution patterns of aPSC (magenta stained with the PIWI-A antibodies).

Now, we are investigating differences of these cellularsystems between highly- and lower- regenerative animals.

Publication List:〔Original papers〕

T., Okumura, M., Bando, Y., Hagiwara, M., Hamada, Y.,• Bando,Ishimaru, Y., Mito, T., Kawaguchi, E., Inoue, T., Agata, K., Noji, S., and•Trials for evoking functional joint regenerationin mouse

Mouse can’t regenerate joint structures at all. However, weexpect that functional joints might be evoked by the activation of reintegration systems since we found that the remaining tissues after amputation at the joint level possessed theability to regenerate joint tissues (Hotta et al., unpublisheddata). That is, the mouse might lose their regenerative abilities due to non-cell autonomous reasons. Therefore, we are
now investigating how to activate reintegration systems inmice after amputation at the joint level for evoking functional joint regeneration.

Isolation of the viable planarian aPSC (adultpluripotent stem cells) and characterization ofthese cells at the single cell level.

We tried to develop an isolation method for viable adultpluripotent stem cells (aPSC) from planarians using FACS.
In the previous method, isolated aPSC were unhealthy after40••••

Ohuchi, H. (2022). Toll signalling promotes blastema cell proliferation
during cricket leg regeneration via insect macrophages. Development149, dev199916. DOI: 10.1242/dev.199916
Finet, C., Kassner, V.A., Carvalho, A.B., Chung, H., Day, J.P., Day, S.,
Delaney, E.K., De Re, F.C., Dufour, H.D., Dupim, E., Izumitani, H.F.,
Gauterio, T.B., Justen, J., Katoh, T., Kopp, A., Koshikawa, S., Longdon,B., Loreto, E.L., Nunes, Maria D.S., Raja, K.K.B, Rebeiz, M., Ritchie,M.G., Saakyan, G., Sneddon, T., Teramoto, M., Tyukmaeva, V.,
Vanderlinde, T., Wey, E.E., Werner, T., Williams, T.M., Robe, L.J., Toda,M.J., and Marletaz, F. (2021). DrosoPhyla: Resources for Drosophilid
Phylogeny and Systematics. GENOME Biol. Evol. 13, evab179. DOI:
10.1093/gbe/evab179
Inoue, T., and Agata, K. (2022). Quantification of planarian behaviors.
Dev. Growth & Differ. 64, 16–37. DOI: 10.1111/dgd.12765
Lee, H., Hikasa, K., Umesono, Y., Hayashi, T., Agata, K., and Shibata,N. (2022). Loss of plac8 expression rapidly leads pluripotent stem cells
to enter active state during planarian regeneration. Development 149,dev199449. DOI: 10.1242/dev.199449
Sato, Y., Umesono, Y., Kuroki, Y., Agata, K., and Hashimoto, C. (2022).
Proliferation maintains the undifferentiated status of stem cells: Therole of the planarian cell cycle regulator Cdh1. Dev. Biol. 482, 55–66.
DOI: 10.1016/j.ydbio.2021.12.006
Takeuchi, T., Matsubara, H., Minamitani, F., Satoh, Y., Tozawa, S.,Moriyama, T., Maruyama, K., Suzuki, K.T., Shigenobu, S., Inoue, T.,Tamura, K., Agata, K., and Hayashi, T. (2022). Newt Hoxa13 has an
essential and predominant role in digit formation during development
and regeneration. Development 149, dev200282. DOI: 10.1242/dev.200282