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The Journal of Neuroscience, October 22, 2003, 23(29):9575-9584

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Development/Plasticity/Repair
Ontogeny of the Pre-Bötzinger Complex in Perinatal Rats

Silvia Pagliardini, Jun Ren, and John J. Greer

Department of Physiology, Centre for Neuroscience, University of Alberta, Edmonton, Alberta, Canada T6G 2S2

The aim of this study was to provide a systematic examination of the ontogenesis of the mammalian respiratory rhythm generating center, the pre-Bötzinger complex (pre-BötC). A combination of immunohistochemical markers and electrophysiological recordings was used to determine the time of inception of the pre-BötC and the developmental changes during the perinatal period in rats spanning from embryonic day 15 (E15) to postnatal day 7. The first clear indication of neurons immunopositive for neurokinin-1 receptors (NK1Rs) and somatostatin expression, two proposed markers for pre-BötC neurons, was at E17. Birth dating of neurons in the ventrolateral medulla using 5-bromo-2'-deoxyuridine demonstrated that NK1R-positive neurons populating the area of the pre-BötC during late E16-E18 are born at E12.5-E13.5, 2 d later than adjacent NK1R-positive neurons in the ventrolateral medulla. Extracellular recordings of neuronal populations within the pre-BötC of perinatal medullary slice preparations demonstrated that the onset of rhythmical respiratory discharge commences at E17. Application of substance P, a ligand for NK1R receptors, to the media bathing E17 medullary slice and brainstem-spinal cord preparations resulted in a marked increase in respiratory frequency. These data provide insights into the ontogeny of the pre-BötC, giving fundamental information on the genesis, settlement, and inception of rhythmic activity within the group of neurons proposed to be responsible for the respiratory rhythm generation. Furthermore, this provides the foundation for further analyses of cell lineage, the transcriptional control of respiratory neuronal development, and electrophysiological and pharmacological properties of the pre-BötC during the prenatal period.

Key words: pre-Bötzinger complex; breathing; perinatal; medulla; respiration; neurokinin-1 receptor

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Received May 15, 2003; revised August 18, 2003; accepted August 22, 2003.

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