The Journal of Physiological Sciences

, Volume 69, Issue 1, pp 65–77 | Cite as

Functional emergence of a column-like architecture in layer 5 of mouse somatosensory cortex in vivo

  • Kyo Koizumi
  • Masatoshi Inoue
  • Srikanta Chowdhury
  • Haruhiko Bito
  • Akihiro Yamanaka
  • Toru Ishizuka
  • Hiromu YawoEmail author
Original Paper


To investigate how the functional architecture is organized in layer 5 (L5) of the somatosensory cortex of a mouse in vivo, the input–output relationship was investigated using an all-optical approach. The neural activity in L5 was optically recorded using a Ca2+ sensor, R-CaMP2, through a microprism inserted in the cortex under two-photon microscopy, while the L5 was regionally excited using optogenetics. The excitability was spread around the blue-light irradiated region, but the horizontal propagation was limited to within a certain distance (λ < 130 μm from the center of the illumination spot). When two regions were photostimulated with a short interval, the excitability of each cluster was reduced. Therefore, a column-like architecture had functionally emerged with reciprocal inhibition through a minimal number of synaptic relays. This could generate a synchronous output from a region of L5 with simultaneous enhancement of the signal-to-noise ratio by silencing of the neighboring regions.


Optogenetics Ca2+ imaging Two-photon microscopy Self-organization Column Reciprocal inhibition 



We thank K. Fukunaga for the generous gift of rabbit pan CaMKII antiserum and B. Bell for language assistance. This work was supported by a Grant-in-Aid for Scientific Research (No. 25250001 to HY), a Grant-in-Aid for Scientific Research on Innovative Areas (Adaptive Circuit Shift: No. 15H01413 to HY; Comprehensive Brain Science Network; Platforms for Advanced Technologies and Research Resources: No. 16H06276 to HB) of the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan: as well as JST, Strategic International Collaborative Research Program, SICORP (to HY): and CREST (JPMJCR1656 to AY).

Author contributions

KK contributed to conceiving, designing and performing the experiments. KK and HY contributed to data analysis. MI, SC, HB, AY and TI contributed by providing reagents, materials, and analysis tools. KK and HY contributed to data interpretation and manuscript writing. All authors approved the final version of the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Statement on the welfare of animals

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted. This article does not contain any studies with human participants performed by any of the authors.

Supplementary material

12576_2018_618_MOESM1_ESM.pdf (31 kb)
Supplementary Figure 1 Circuit diagram for the LED analog regulator. (PDF 30 kb)
12576_2018_618_MOESM2_ESM.pdf (485 kb)
Supplementary Figure 2 Cortical organization. The neurons expressing R-CaMP2 in layer 2/3 and 5 were imaged through a microprism under two-photon microscopy. Scale, 100 µm. (PDF 485 kb)
12576_2018_618_MOESM3_ESM.avi (1.8 mb)
Supplementary Video 1 A sample R-CaMP2 fluorescence image at layer 5 through the prism (the same region shown in Figure 3A). The targets of photostimulation, A, B, and C are overlaid (circles). The illuminated targets are colored in blue in the frame capturing the LED-dependent noise at the instant of photostimulation (AVI 1836 kb)


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Copyright information

© The Physiological Society of Japan and Springer Japan KK, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Developmental Biology and NeuroscienceTohoku University Graduate School of Life SciencesSendaiJapan
  2. 2.Department of Neurochemistry, Graduate School of MedicineUniversity of TokyoTokyoJapan
  3. 3.Core Research for Evolutional Science and Technology (CREST)Japan Science and Technology AgencyTokyoJapan
  4. 4.Department of Neuroscience II, Research Institute of Environmental MedicineNagoya UniversityNagoyaJapan
  5. 5.Center for NeuroscienceTohoku University Graduate School of MedicineSendaiJapan

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