Vana Košta

Characterization of spinal afferent neurons projecting to different chambers of the rat heart

The pattern of distribution of spinal afferent neurons (among dorsal root ganglia-DRGs) that project to anatomically and functionally different chambers of the rat heart, as well as their morphological and neurochemical characteristics were investigated. Retrograde tracing using a patch loaded with Fast blue (FB) was applied to all four chambers of the rat heart and labeled cardiac spinal afferents were characterized by using three neurochemical markers. The majority of cardiac projecting neurons were found from T1 to T4 DRGs, whereas the peak was at T2 DRG. There was no difference in the total number of FB-labeled neurons located in ipsilateral and contralateral DRGs regardless of the chambers marked with the patch. However, significantly more FB-labeled neurons projected to the ventricles compared to the atria (859 vs. 715). The proportion of isolectin B(4) binding in FB-labeled neurons was equal among all neurons projecting to different heart chambers (2.4%). Neurofilament 200 positivity was found in greater proportions in DRG neurons projecting to the left side of the heart, whereas calretinin-immunoreactivity was mostly represented in neurons projecting to the left atrium. Spinal afferent neurons projecting to different chambers of the rat heart exhibit a variety of neurochemical phenotypes depending on binding capacity for isolectin B(4) and immunoreactivity for neurofilament 200 and calretinin, and thus represent important baseline data for future studies.

Immunohistochemical characteristics of neurons in nodose ganglia projecting to the different chambers of the rat heart

Despite the contribution of nodose ganglia neurons to the innervation of the heart being the subject of several studies, specific neuronal subpopulations innervating the four different chambers of the heart have not been distinguished. In our study, the application of Fast Blue-loaded patch to the epicardial surface of different chambers of the rat heart (the right or left atrium or the right or left ventricle) resulted in labeling of discrete populations of immunohistochemically diverse neurons. About one half (55%) of these neurons showed immunoreactivity for the 200-kDa neurofilament protein (marker of myelinated neurons), with a higher proportion of positive staining among neurons projecting to the left than to the right ventricle. Isolectin B4 immunoreactivity (characteristic for a subset of nonmyelinated non-peptidergic neurons) was more abundant among neurons projecting to the right side of the heart (right atria and right ventricles) compared to the left side (23% vs. 16%). Calretinin immunoreactivity (possible marker of mechanosensitive neurons) was significantly higher among neurons projecting to the ventricles than among those projecting to atria (36% vs. 11%). These findings reveal that chambers of the rat heart are innervated with immunohistochemically different subpopulations of neurons from the nodose ganglia.

The extent of laminectomy affects pain-related behavior in a rat model of neuropathic pain

One of the unresolved questions in neuropathic pain research is whether we can prevent or reverse mechanical hyperalgesia by rhizotomy or ganglionectomy. However, one of the obstacles in answering that question is lack of a standardized surgical procedure used in experimental ganglionectomy. We tested the hypothesis that laminectomy performed during ganglionectomy induces lumbar column deformity. We further examined whether spinal deformity is a source of pain‐related behavior. Five conditions were studied. Fifth and sixth lumbar (L5 and L6) ganglionectomy were performed in rats using either minimal or extensive laminectomy technique. Two other groups had minimal and extensive laminectomy without ganglionectomies. A final control group had no surgery. Sensory responsiveness of the plantar aspect of the hind paw was repeatedly tested, and a plain radiograph in anteroposterior projection was made to assess the extent of deformity by measurement of deformity angles. Hyperalgesia resulted in groups with extensive laminectomy regardless of performance or absence of ganglionectomy, while in groups with minimal laminectomy there was no increase in pain‐related behavior. Lateral deformity of the spine was observed in rats with or without ganglionectomy, confirming that laminectomy can produce deformity. The extent of deformity was more pronounced in rats exposed to the extensive laminectomy. Our results indicate that laminectomy can produce spine deformity and that there is a direct relationship between the extent of laminectomy and the development of mechanical hypersensitivity. The data presented suggest that there is a need for standardization of laminectomy procedure in rat experimental pain models.

The influence of exercise on morphological and neurochemical properties of neurons in rat nodose ganglia

Physical exercise can induce immunohistochemical changes and cell proliferation in the hippocampus. One of the main effects of prolonged exercise is resting bradycardia, most probably caused by enhanced vagal activity. To investigate whether physical exercise can cause neurochemical and morphological changes in vagal afferent neurons, we performed immunohistochemical studies of nodose neurons using isolectin B4 (IB4), 200-kDa neurofilament protein (N52) and calretinin in adult female rats. To distinguish subpopulations of neurons projecting to the left ventricle, we applied a Fast Blue patch to the epicardial surface of the left ventricle. Treadmill running for 8 weeks significantly increased the size of N52-positive cardiac projecting neurons. Furthermore, the proportion of IB4-positive neurons among all nodose ganglia neurons was significantly higher in trained animals. These data indicate that exercise leads to plastic changes in nodose ganglia neurons that may initiate changes of vagal activity caused by prolonged exercise.

Postlaminectomy stabilization of the spine in a rat model of neuropathic pain reduces pain-related behavior

Study Design. Spine deformity and pain-related behavior after laminectomy with and without spine stabilization were investigated. Objective. We tested hypothesis that spine stabilization after extensive laminectomy can prevent spine deformation and consequent pain-related behavior. Summary of Background Data. Various ablative procedures requiring laminectomy have been tested for prevention or reversal of pain-related behavior in studies using experimental animals. However, there is no precise description indicating how laminectomy should be performed. Lack of standardized surgical techniques makes it difficult to achieve uniformity of result reporting and to compare results of different research groups meaningfully. Methods. To test our hypothesis, extensive laminectomy with and without spine stabilization was performed in Sprague-Dawley rats. U-shaped surgical wire was used for stabilization of the spine. A validated test of mechanical hyperalgesia was used to test the development of neuropathic pain behavior after surgery. Deformity of the spine was evaluated by calculating deviation from the central axis on radiographs obtained in anteroposterior projection. Results. Surgical stabilization of the spine after laminectomy prevented development of spinal deformity. Laminectomy without stabilization induced hyperalgesia on the 8th and 15th days after surgery. Group with stabilized spine exhibited significant reduction in pain-related behavior on the 8th and 15th postoperative days compared with the group without stabilization. Conclusion. Surgical stabilization of the spine after laminectomy prevented development of spinal deformity and pain-related behavior. Our results suggest that spine stabilization procedure should be used in all experimental pain models in which laminectomy is performed.

Age-related changes of neurochemically different subpopulations of cardiac spinal afferent neurons in rats.

This study investigated the effect of aging on cardiac spinal afferent neurons in the rat. A patch loaded with retrograde tracer Fast Blue (FB) was applied to all chambers of the rat heart. Morphological and neurochemical characteristics of labeled cardiac spinal afferent neurons were assessed in young (2 months) and old (2 years) rats using markers for likely unmyelinated (isolectin B4; IB4) and myelinated (neurofilament 200; N52) neurons. The number of cardiac spinal afferent neurons decreased in senescence to 15% of that found in young rats (1604 vs. 248). The size of neuronal soma as well as proportion of IB4+ neurons increased significantly, whereas the proportion of N52+ neurons decreased significantly in senescence. Unlike somatic spinal afferents, neurochemically different populations of cardiac spinal afferent neurons experience morphological and neurochemical changes related to aging. A major decrease in total number of cardiac spinal afferent neurons occurs in senescence. The proportion of N52+ neurons decreased in senescence, but it seems that nociceptive innervation is preserved due to increased proportion and size of IB4+ unmyelinated neurons.

165 SPINE DEFORMATION IN RAT MODELS OF NEUROPATHIC PAIN CAN CAUSE PAIN RELATED BEHAVIOR

administration. The reversal of tactile allodynia was maintained with repeated administration for 5 days. Similar reversal of tactile allodynia was seen in a model of vincristine-induced neuropathic pain. In addition, long term treatment with TRO19622 normalized motor nerve conduction in diabetic rats suggesting that this compound may improve neuropathic syndromes in addition to analgesic activity. By contrast, TRO19622 did not reverse thermal hyperalgesia in normal or neuropathic rats or pain-like behaviour in the formalin test. These data suggest that TRO19622 specifically reverses neuropathic rather than noxious or inflammatory pain. Even at very high doses TRO19622 is devoid of sedative or other adverse effects of current analgesics and therefore offers a promising new approach for treating neuropathic pain. A Phase 2 study in diabetic neuropathic pain is currently ongoing.