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Activity-dependent homeostatic specification of transmitter expression in embryonic neurons

Abstract

Neurotransmitters are essential for interneuronal signalling, and the specification of appropriate transmitters in differentiating neurons has been related to intrinsic neuronal identity and to extrinsic signalling proteins. Here we show that altering the distinct patterns of Ca2+ spike activity spontaneously generated by different classes of embryonic spinal neurons in vivo changes the transmitter that neurons express without affecting the expression of markers of cell identity. Regulation seems to be homeostatic: suppression of activity leads to an increased number of neurons expressing excitatory transmitters and a decreased number of neurons expressing inhibitory transmitters; the reverse occurs when activity is enhanced. The imposition of specific spike frequencies in vitro does not affect labels of cell identity but again specifies the expression of transmitters that are inappropriate for the markers they express, during an early critical period. The results identify a new role of patterned activity in development of the central nervous system.

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Figure 1: Ca2+ spike activity of four classes of neurons imaged in the embryonic spinal cord.
Figure 2: Suppression of spike activity in vivo by overexpression of inward rectifier K+ channels increases the incidence of expression of glutamatergic and cholinergic phenotypes.
Figure 3: Enhancement of spike activity in vivo by overexpression of voltage-gated Na+ channels decreases the incidence of glutamatergic and cholinergic phenotypes. rNav2aαβ transcripts and fluorescent tracer were injected together into one or both blastomeres at the two-cell stage, followed by imaging and immunocytochemistry as in Fig. 2.
Figure 4: Pharmacological in vivo suppression of spikes with Ca2+ and Na+ channel blockers or enhancement of spikes with the Na+ channel agonist veratridine enhances or suppresses Glu-IR and ChAT-IR, respectively.
Figure 5: Suppression or enhancement of spike activity in vivo causes homeostatic superposition or replacement of one transmitter with another.
Figure 6: Regulation of spike frequency in vitro drives novel expression of neurotransmitters.
Figure 7: Neurotransmitters expressed after alterations in Ca2+ spike activity are functionally released.
Figure 8: Critical periods for Ca2+ spike-dependent regulation of transmitter expression in vitro.

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Acknowledgements

We thank D. Berg, D. Feldman, M. Feller and A. Ghosh for comments on the manuscript, T. Jessell for discussions, and I-T. Hsieh for technical support. This work was supported by a grant to N.C.S. from the National Institutes of Health. S.B.S. was supported by the NSF and Merck.

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Correspondence to Laura N. Borodinsky.

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Supplementary information

Supplementary Figure 1

Coactively spiking clusters of MNs in the embryonic neural tube.

Supplementary Figure 2

The cellular organization of the neural tube appears normal following bilateral suppression of Ca2+ spike activity.

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Borodinsky, L., Root, C., Cronin, J. et al. Activity-dependent homeostatic specification of transmitter expression in embryonic neurons. Nature 429, 523–530 (2004). https://doi.org/10.1038/nature02518

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