Journal of Physiology-London
Key points center dot The transcription factor Dbx1 gives rise to putatively respiratory rhythm-generating neurons in the pre-Botzinger complex. Comparative analysis of Dbx1-derived (Dbx1+) and non-Dbx1- derived (Dbx1) neurons can help elucidate the cellular bases of respiratory rhythm generation. center dot In vitro, Dbx1+ neurons activate earlier in the respiratory cycle, discharge larger magnitude inspiratory bursts and exhibit a lower rheobase compared with Dbx1 neurons. center dot The Dbx1+ neurons tend to express the intrinsic currents IA (transient outward A-current) and Ih (hyperpolarization-activated current) in diametric opposition, which may facilitate temporal summation of excitatory synaptic inputs, whereas the Dbx1 neurons show no significant pattern of expression regarding IA and Ih. center dot The Dbx1+ neurons exhibit smooth, spineless dendrites that project in the transverse plane, whereas the Dbx1 neurons are confined to the transverse plane to a lesser extent and sometimes exhibit spines. center dot The properties of Dbx1+ neurons that may contribute to respiratory rhythmogenesis include a high level of excitability linked to ongoing network activity and dendritic properties that may facilitate synaptic integration. Abstract Breathing in mammals depends on an inspiratory-related rhythm that is generated by glutamatergic neurons in the pre-Botzinger complex (preBotC) of the lower brainstem. A substantial subset of putative rhythm-generating preBotC neurons derive from a single genetic line that expresses the transcription factor Dbx1, but the cellular mechanisms of rhythmogenesis remain incompletely understood. To elucidate these mechanisms, we carried out a comparative analysis of Dbx1-expressing neurons (Dbx1+) and non-Dbx1-derived (Dbx1) neurons in the preBotC. Whole-cell recordings in rhythmically active newborn mouse slice preparations showed that Dbx1+ neurons activate earlier in the respiratory cycle and discharge greater magnitude inspiratory bursts compared with Dbx1 neurons. Furthermore, Dbx1+ neurons required less input current to discharge spikes (rheobase) in the context of network activity. The expression of intrinsic membrane properties indicative of A-current (IA) and hyperpolarization-activated current (Ih) tended to be mutually exclusive in Dbx1+ neurons. In contrast, there was no such relationship in the expression of currents IA and Ih in Dbx1 neurons. Confocal imaging and digital morphological reconstruction of recorded neurons revealed dendritic spines on Dbx1 neurons, but Dbx1+ neurons were spineless. The morphology of Dbx1+ neurons was largely confined to the transverse plane, whereas Dbx1 neurons projected dendrites to a greater extent in the parasagittal plane. The putative rhythmogenic nature of Dbx1+ neurons may be attributable, in part, to a higher level of intrinsic excitability in the context of network synaptic activity. Furthermore, Dbx1+ neuronal morphology may facilitate temporal summation and integration of local synaptic inputs from other Dbx1+ neurons, taking place largely in the dendrites, which could be important for initiating and maintaining bursts and synchronizing activity during the inspiratory phase.
Guinamard, R., Simard, C., & Del Negro, C. (2013). Flufenamic acid as an ion channel modulator. Pharmacology & therapeutics, 138(2), 272-284.