TY - JOUR
T1 - Reelin transiently promotes N-cadherin-dependent neuronal adhesion during mouse cortical development
AU - Matsunaga, Yuki
AU - Noda, Mariko
AU - Murakawa, Hideki
AU - Hayashi, Kanehiro
AU - Nagasaka, Arata
AU - Inoue, Seika
AU - Miyata, Takaki
AU - Miura, Takashi
AU - Kubo, Ken Ichiro
AU - Nakajima, Kazunori
N1 - Funding Information:
We thank Dr. T. Curran, Dr. J. Miyazaki, Dr. J. Takagi, Dr. R. L. Huganir, and Dr. A. Nagafuchi for reagents and advice. We thank the members of the K.N. laboratory for valuable discussions and technical support. This work was supported by the Ministry of Education, Culture, Sports, Science and Technology (MEXT)/Japan Society for the Promotion of Science Grantsin-Aid for Scientific Research (KAKENHI) (Grants JP16H06482, JP15H02355, JP15H01586, JP16H01343, JP16J05704, JP15H01293, JP26430075, JP15K09723, JP16K09997, JP15K06745, and JP26400205), Takeda Science Foundation, Naito Foundation, Keio Gijuku Academic Development Funds, Keio University Grant-in-Aid for Encouragement of Young Medical Scientists, Program for the Advancement of Keio Next Generation Research Projects, and Fukuzawa Memorial Fund for the Advancement of Education and Research.
Publisher Copyright:
© 2017, National Academy of Sciences. All rights reserved.
PY - 2017/2/21
Y1 - 2017/2/21
N2 - Reelin is an essential glycoprotein for the establishment of the highly organized six-layered structure of neurons of the mammalian neocortex. Although the role of Reelin in the control of neuronal migration has been extensively studied at the molecular level, the mechanisms underlying Reelin-dependent neuronal layer organization are not yet fully understood. In this study, we directly showed that Reelin promotes adhesion among dissociated neocortical neurons in culture. The Reelin-mediated neuronal aggregation occurs in an N-cadherin-dependent manner, both in vivo and in vitro. Unexpectedly, however, in a rotation culture of dissociated neocortical cells that gradually reaggregated over time, we found that it was the neural progenitor cells [radial glial cells (RGCs)], rather than the neurons, that tended to form clusters in the presence of Reelin. Mathematical modeling suggested that this clustering of RGCs could be recapitulated if the Reelin-dependent promotion of neuronal adhesion were to occur only transiently. Thus, we directly measured the adhesive force between neurons and N-cadherin by atomic force microscopy, and found that Reelin indeed enhanced the adhesiveness of neurons to N-cadherin; this enhanced adhesiveness began to be observed at 30 min after Reelin stimulation, but declined by 3 h. These results suggest that Reelin transiently (and not persistently) promotes N-cadherin-mediated neuronal aggregation. When N-cadherin and stabilized β-catenin were overexpressed in the migrating neurons, the transfected neurons were abnormally distributed in the superficial region of the neocortex, suggesting that appropriate regulation of N-cadherin-mediated adhesion is important for correct positioning of the neurons during neocortical development.
AB - Reelin is an essential glycoprotein for the establishment of the highly organized six-layered structure of neurons of the mammalian neocortex. Although the role of Reelin in the control of neuronal migration has been extensively studied at the molecular level, the mechanisms underlying Reelin-dependent neuronal layer organization are not yet fully understood. In this study, we directly showed that Reelin promotes adhesion among dissociated neocortical neurons in culture. The Reelin-mediated neuronal aggregation occurs in an N-cadherin-dependent manner, both in vivo and in vitro. Unexpectedly, however, in a rotation culture of dissociated neocortical cells that gradually reaggregated over time, we found that it was the neural progenitor cells [radial glial cells (RGCs)], rather than the neurons, that tended to form clusters in the presence of Reelin. Mathematical modeling suggested that this clustering of RGCs could be recapitulated if the Reelin-dependent promotion of neuronal adhesion were to occur only transiently. Thus, we directly measured the adhesive force between neurons and N-cadherin by atomic force microscopy, and found that Reelin indeed enhanced the adhesiveness of neurons to N-cadherin; this enhanced adhesiveness began to be observed at 30 min after Reelin stimulation, but declined by 3 h. These results suggest that Reelin transiently (and not persistently) promotes N-cadherin-mediated neuronal aggregation. When N-cadherin and stabilized β-catenin were overexpressed in the migrating neurons, the transfected neurons were abnormally distributed in the superficial region of the neocortex, suggesting that appropriate regulation of N-cadherin-mediated adhesion is important for correct positioning of the neurons during neocortical development.
KW - Aggregation
KW - Corticogenesis
KW - N-cadherin
KW - Neuronal migration
KW - Reelin
UR - http://www.scopus.com/inward/record.url?scp=85013269752&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85013269752&partnerID=8YFLogxK
U2 - 10.1073/pnas.1615215114
DO - 10.1073/pnas.1615215114
M3 - Article
C2 - 28174271
AN - SCOPUS:85013269752
SN - 0027-8424
VL - 114
SP - 2048
EP - 2053
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 8
ER -