Stationary Organotypic Culture of the Pre-Bötzinger Complex from the Newborn Rat

Abstract

In the early 1990s, it was suggested that the central neuronal network responsible for breathing relies on pacemaker cells to promote its rhythmic activity.^1,2,3These pacemaker neurons were shown to be concentrated in the region of the pre-Bötzinger complex and are critical components of the network generating respiratory rhythm.^3,4Previous studies of this neuronal population have been done primarily in fresh tissue slices in vitro which have a limited survival (less than 48 hours) and, due to thickness of the slice, offer poor accessibility to neurons of interest. Tissue culture of the isolated pre-Bötzinger complex may offer some advantages in this respect, but it is not known whether the isolated pre-Bötzinger network would maintain its functional properties. Therefore, we evaluated the viability of the stationary explant of the cultured pre-Bötzinger region (“island”). We hypothesized that the pacemaker network would maintain functional activity for extended periods in culture due to the network robust functional properties.