Immunocytochemical localization of glutamic acid decarboxylase in physiologically identified interneurons of hamster spinal laminae III-V.

Neurons in Rexed's laminae III-V of an isolated spinal cord-skin patch preparation from hamsters were recorded in whole-cell mode and stained intracellularly with biocytin. Evidence of inhibitory synaptic function was obtained via post-hoc immunofluorescent labeling with a monoclonal antibody directed against an axon terminal isoform of brain glutamic acid decarboxylase. For a subset of neurons, examination with laser scanning confocal microscopy revealed punctate accumulations of glutamic acid decarboxylase immunoreactivity within axon enlargements (1-3 microm diameter), as imaged in single optical sections and confirmed by subsequent optical scans in the orthogonal plane. Axons of glutamic acid decarboxylase-immunoreactive neurons were found to exhibit dense local terminations overlapping the soma and dendrites or bifurcated into lengthy rostrocaudal daughter branches ventral to the cell body. The degree and uniformity of immunolabeling in axonal enlargements varied considerably, even amongst boutons belonging to the same cell. Glutamic acid decarboxylase-positive neurons received input from myelinated (A) afferent fibers and responded to natural stimuli appropriate for activating responses in low threshold mechanoreceptors. These results provide evidence that two different populations of GABAergic inhibitory interneurons are involved in local and intersegmental circuits that mediate integration of mechanosensory information in the deep spinal dorsal horn.