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Received November 28, 2007
Revised January 22, 2008
Accepted after revision February 26, 2008
1 Department of Neuro- and Sensory Physiology, Georg-August-University, 37073 Göttingen, Germany
* To whom correspondence should be addressed. E-mail: smirono{at}gwdg.de.
Respiration in the vertebrates is generated by a compact network which is located in the lower brainstem but cellular mechanisms which underlie persistent oscillatory activity of the respiratory network are yet unknown. Using two-photon imaging and patch-clamp recordings in functional brainstem preparation of mice containing pre-Bötzinger complex (preBötC), we examined the actions of metabotropic glutamate receptors (mGluR1/5) on the respiratory patterns. The agonist DHPG potentiated and antagonist LY367385 depressed respiration-related activities. In the inspiratory neurons, we observed rhythmic activation of non-selective channels which had conductance 24 pS. Their activity was enhanced with membrane depolarisation and after elevation of calcium from the cytoplasmic side of the membrane. They were activated by non-hydrolysable PIP2 analogue and blocked by flufenamate, ATP4- and Gd3+. All these properties correspond well to those of TRPM4 channels. Calcium imaging of functional slices revealed rhythmic transients in small clusters of neurones present in a network. Calcium transients in the soma were preceded by the waves in dendrites which were dependent on mGluR activation. Initiation and propagation of waves required calcium influx and calcium release from internal stores. Calcium waves activated TPRM4-like channels in the soma and promoted generation of inspiratory bursts. Simulations of activity of neurons communicated via dendritic calcium waves showed emerging activity within neuronal clusters and its synchronisation between the clusters. The experimental and theoretical data provide a subcellular basis for a recently proposed group-pacemaker hypothesis and describe a novel mechanism of rhythm generation in neuronal networks.
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