Spinal Cord Stimulation and the Induction of c-fos and Heat Shock Protein 72 in the Central Nervous System of Rats

For more than a decade, spinal cord stimulation (SCS) has been used as an adjuvant treatment for patients who are unresponsive to conventional therapies for angina pectoris. Many studies showed that SCS has both electro-analgesic and anti-ischemic effects. Nonetheless, the biological substrates by which SCS acts have not yet been unraveled, although recently areas in the brain have been described that show changes in blood flow, following SCS, and during provocation of angina. In search of a putative mechanism of action of SCS, we hypothesized that SCS affects processing of nociceptive information within the central nervous system (CNS). Moreover, it may alter the limbic system activity that maintains the balance between sympathetic and parasympathetic activity in the heart. Hence, we have developed a rat model to investigate its suitability for studying the induction of neural activity during SCS. To characterize neural activity, we used the expression of both the immediate early gene c-fos and the heat shock protein 72 (HSP72). c-Fos was used to identify structures in the CNS affected by SCS, and HSP72 was applied in order to ascertain whether SCS might operate as a stressor. In 20 halothane-anesthetized male Wistar rats, two electrodes were placed epidurally, one at the C7 level and the other at the T2 level. Two days after surgery, the rats were either stimulated "treated" animals, n = 10) or used as controls ("unstimulated" = "sham," n = 10) in random order. Furthermore, we studied the effect of SCS on behavior in five treated and five control rats. Three hours after stimulation, the rats were euthanized and the brain and spinal cord were removed. The treated group showed regional increased c-fos expression in regions of the limbic system (periaqueductal gray, paraventricular hypothalamic nucleus, paraventricular thalamic nucleus, central amygdala, agranular and dysgranular insular cortex, (peri)ambiguus, nucleus tractus solitarius, and spinal cord) that are involved in the processing of pain and cardiovascular regulation, among other things. Moreover, in both treated rats and controls, HSP72-expression was found in the endothelium of the enthorhinal cortex, the amygdala, and the ventral hypothalamus, but not in the neurons. Finally, treated animals were significantly more alert and active than controls. In conclusion, the rat model we developed appears to be suitable for studying potential mechanisms through which SCS may act. In addition, SCS affects c-fos expression in specific parts of the brain known to be involved in regulation of pain and emotions. HSP72-expression is limited to the endothelium of certain parts of the CNS and thereby excludes physical stress effects as a potential mechanism of SCS.