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Specific synapses develop preferentially among sister excitatory neurons in the neocortex


Neurons in the mammalian neocortex are organized into functional columns1,2. Within a column, highly specific synaptic connections are formed to ensure that similar physiological properties are shared by neuron ensembles spanning from the pia to the white matter. Recent studies indicate that synaptic connectivity in the neocortex is sparse and highly specific3,4,5,6,7,8 to allow even adjacent neurons to convey information independently9,10,11,12. How this fine-scale microcircuit is constructed to create a functional columnar architecture at the level of individual neurons largely remains a mystery. Here we investigate whether radial clones of excitatory neurons arising from the same mother cell in the developing neocortex serve as a substrate for the formation of this highly specific microcircuit. We labelled ontogenetic radial clones of excitatory neurons in the mouse neocortex by in utero intraventricular injection of enhanced green fluorescent protein (EGFP)-expressing retroviruses around the onset of the peak phase of neocortical neurogenesis. Multiple-electrode whole-cell recordings were performed to probe synapse formation among these EGFP-labelled sister excitatory neurons in radial clones and the adjacent non-siblings during postnatal stages. We found that radially aligned sister excitatory neurons have a propensity for developing unidirectional chemical synapses with each other rather than with neighbouring non-siblings. Moreover, these synaptic connections display the same interlaminar directional preference as those observed in the mature neocortex. These results indicate that specific microcircuits develop preferentially within ontogenetic radial clones of excitatory neurons in the developing neocortex and contribute to the emergence of functional columnar microarchitectures in the mature neocortex.

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Figure 1: Morphological development of ontogenetic radial clones of excitatory neurons in the neocortex.
Figure 2: Synapse formation between sister excitatory neurons in ontogenetic radial clones.
Figure 3: A strong preference for synapse formation between sister excitatory neurons in ontogenetic radial clones.
Figure 4: A highly specific microcircuit forms among sister excitatory neurons in ontogenetic radial clones.


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We thank A. L. Joyner, J. Kaltschmidt, Y. Hayashi and Y. Chin for comments on the manuscript; and S. C. Noctor and F. H. Gage for providing the 293gp NIT–GFP retrovirus packaging cell line; and the Shi laboratory members for insightful discussion. We are grateful for support from the March of Dimes Foundation, the Whitehall Foundation, the Klingenstein Foundation, the DANA Foundation, the Autism Speaks Foundation, the National Alliance for Research on Schizophrenia and Depression (NARSAD) and the National Institutes of Health (to S.-H.S.).

Author Contributions Y.-C.Y. and S.-H.S. conceived the experiments. Y.-C.Y. conducted the electrophysiology and imaging experiments. R.S.B and X.W. helped with in utero virus injection. Y.-C.Y. and S.-H.S. analysed the data and wrote the manuscript.

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Correspondence to Song-Hai Shi.

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Yu, YC., Bultje, R., Wang, X. et al. Specific synapses develop preferentially among sister excitatory neurons in the neocortex. Nature 458, 501–504 (2009).

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