Selective labelling of primary sensory afferent terminals in lamina II of the dorsal horn by injection of Bandeiraea simplicifolia isolectin B4 into peripheral nerves

The I-B4 isolectin from Bandeiraea simplicifolia exhibits specific binding to a subpopulation of rat dorsal root ganglion neurons of small diameter which terminate in the substantia gelatinosa of the dorsal horn. Recent double-labelling experiments in the rat have demonstrated that only primary afferents which innervate the skin are recognized by the I-B4 lectin [Plenderleith and Snow (1993) Neurosci. Lett. (in press)]. As the I-B4 lectin appears to bind selectively to a subset of small-diameter primary afferents with cutaneous peripheral projections, we sought to determine whether it could be used as a transganglionic tracer which selectively labels the spinal terminations of cutaneous afferents in superficial dorsal horn. We now report that the I-B4-horseradish peroxidase conjugate labels synaptic terminals in lamina II of the dorsal horn following the injection of the conjugate into the sciatic and saphenous nerves in the rat. Electron-microscopic examination of the dorsal horn revealed many examples of labelled synaptic terminals and unmyelinated axons, but in no cases was label observed in myelinated axons. No label was observed outside of the substantia gelatinosa; thus the I-B4 isolectin is unique among lectins used for transganglionic tracing in that it does not retrogradely label motoneurons. These results, together with previous studies of lectin binding properties of primary sensory afferents, suggest that injection of I-B4 conjugates into peripheral nerves enables the visualization of the central terminations of cutaneous C-fibres. Transganglionic labelling with the I-B4 isolectin from Bandeiraea simplicifolia should facilitate further examination of synaptic relationships of nociceptive cutaneous afferents in the superficial dorsal horn