Modulation of substance P-ergic system in the rat spinal cord by an opioid antagonist

Abstract

Substance P- and opioid peptide-immunoreactive nerve terminals functionally interact in the spinal cord as two opposing systems in the regulation of the nociceptive pathway. In order to determine how SP-ergic system adapts to chronic opioid receptor blockade, the effects of naltrexone on SP level, SP receptor and the second messenger system coupled to the SP receptor were examined in the rat spinal cord. Male Sprague-Dawley rats were treated with naltrexone or vehicle for seven days by constant minipump infusion. Animals were sacrificed on day 8, spinal cords rapidly removed, segmentally sectioned and used to determine SP and inositol 1,4,5-trisphosphate [ins(1,4,5)P 3] tissue contents, and to examine the regulation of their respective receptors in in vitro receptor binding assays. Following chronic naltrexone treatment, SP content in the lumbosacral segment of the spinal cord was increased by 53% over matched control values. The binding capacity ( B max) of SP receptors, determined using [ 125I]BHSP, in lumbosacral synaptosomal membranes was significantly increased by 92%, but the binding affinity ( K d) remained unchanged. In addition, the concentration of [Sar 9, Met(O 2) 11]SP, an NK-1 receptor-specific agonist, required to inhibit half of [ 125I]BHSP binding (IC 50) in lumbosacral synaptosomal membranes was significantly decreased, but the IC 50s for SP, the endogenous ligand for the SP receptor, and [Pro 7]NK B, an NK-3 receptor-specific agonist, were unaltered by chronic blockade of opioid receptors. The data suggest that although naltrexone does not directly interact with tachykinin receptors, it acts indirectly on SP-ergic neurons to cause a change in the apparent affinity of NK-1 receptor (as reflected by a change in IC 50 value). Formation of cellular ins(1,4,5)P 3 in the lumbosacral cord, quantified by a highly sensitive and selective radioreceptor assay, was significantly increased by 34% relative to matched controls. A time course study indicated that increases in ins(1,4,5)P 3 contents over the time studied corresponded qualitatively with increases in SP level in the lumbosacral cord. With [ 3H]ins(1,4,5)P 3 as a ligand, Scatchard analyses of the concentration dependent saturation curves showed that the density of intracellular ins(1,4,5)P 3 receptors was also increased by 119%, with no change in binding affinity. The data suggest that ins(1,4,5)P 3 formation, possibly coupled to functional SP receptor activation, and ins(1,4,5)P 3 receptors, which mediate ins(1,4,5)P 3-induced alterations in intracellular Ca 2+ flux, are increased in the lumbosacral cord by chronic blockade of opioid receptors. Taken together, the data support the concept of a role for endogenous opioids in the regulation of SP receptor activity in the spinal cord.