Whole-cell patch-clamp recordings from visualized bulbospinal neurons in the brainstem slices

The purpose of this study was to develop a method for electrophysiological characterization of retrogradely labeled bulbospinal neurons in the specific cytoarchitectonic regions in the brainstem slices. Several days after the spinal cord was injected with the carbocyanine dye, DiI, retrogradely labeled bulbospinal neurons were visualized by epifluorescence optics in the brainstem slices with the aid of a silicon intensifier tube (SIT) camera. Labeled somata were routinely seen in the caudal raphe nuclei, rostroventral medial and lateral portions of the medulla, the A5 group and in other medullary sites known to project to the spinal cord. Electrophysiological properties of the DiI-labeled neurons were assessed by whole-cell recordings using micropipettes filled with biocytin. The slices were subsequently processed for dual visualization of biocytin and serotinin or a marker for noradrenergic neurons, tyrosine hydroxylase (TH). The electrophysiological properties of bulbospinal neurons were correlated with their morphology and neurochemical content. This technique may be useful in other areas of CNS for studying morphology, neurochemical content and physiology of retrogradely labeled neurons.