Attenuation of widespread hypersensitivity to noxious mechanical stimuli by inhibition of GABAergic neurons of the right amygdala in a rat model of chronic back pain.

BACKGROUND: Chronic primary low back pain may be associated with hyperalgesia in uninjured tissues and with decreased pain inhibition. Previous studies have shown that the amygdala is involved in pain regulation and chronic pain, that neuronal activity in the amygdala is altered in models of persistent pain, and that the central nucleus of the right amygdala plays an active role in widespread hypersensitivity to noxious stimuli. METHODS: Behavioral, electrophysiological, biochemical, and chemogenetic methods were used to examine the role of the central nucleus of the right amygdala in hypersensitivity to noxious stimuli in a rat model of chronic back pain induced by a local injection of Complete Freund Adjuvant (CFA) in paraspinal muscles. RESULTS: CFA produced chronic inflammation limited to the injected area. CFA-treated rats showed increased pain-like (liking) behaviors during the formalin test compared with controls. They also showed widespread mechanical hypersensitivity compared with controls, which persisted for 2 months. This widespread hypersensitivity was accompanied by altered activity of different types of right amygdala neurons, as shown by extracellular recordings. Plasmatic levels of IL-1ß, IL-6, and TNF-α were not elevated after 1 or 2 months, indicating that persistent widespread hypersensitivity is not caused by persistent systemic inflammation. However, chemogenetic inhibition of GABAergic neurons in the right amygdala attenuated widespread mechanical hypersensitivity. CONCLUSIONS: These findings indicate that chronic widespread mechanical hypersensitivity in a model of chronic back pain can be attenuated by inhibiting GABAergic neurons of the right amygdala, and that widespread hypersensitivity is not maintained by chronic systemic inflammation. SIGNIFICANCE: The amygdala is a key structure involved in pain perception and modulation. The present results indicate that the GABAergic neurons of its central nucleus are involved in widespread hypersensitivity to noxious stimuli in a rat model of chronic back pain. The inhibition of amygdala GABAergic neurons may be a potential target for future interventions in patients with chronic back pain.

Early and late visual deprivation induce hypersensitivity to mechanical and thermal noxious stimuli in the ZRDBA mouse.

BACKGROUND: Visual deprivation leads to behavioural adaptations. Early visual deprivation has greater effects on sensory systems compared with late visual deprivation. Although this has been well studied, the impact of visual deprivation on pain sensitivity has scarcely been investigated. In humans, one study indicates that pain sensitivity is increased in early, but not late-onset blindness. In animals, one study indicates that sensitivity to noxious stimulation is increased in anophthalmic mice, but the impact of late visual deprivation on sensitivity remains unknown. The aim of this behavioural study was to examine sensitivity to noxious stimulation in mice with early and late visual deprivation. We hypothesized that visual deprivation would have different effects on sensitivity to noxious stimulation depending on its onset. METHODS: In Experiment 1, mechanical and thermal sensitivity was examined in four ZRDBA mouse groups: sighted mice, anophthalmic mice, dark-reared sighted mice and adult sighted mice deprived of vision for one week. In Experiment 2, mechanical and thermal sensitivity was examined in adult sighted ZRDBA mice deprived of vision for two months. RESULTS: Anophthalmic and dark-reared mice showed mechanical and thermal hypersensitivity, while the one-week visual deprivation did not alter sensitivity. The two-month deprivation also resulted in mechanical and thermal hypersensitivity. CONCLUSIONS: These results indicate that early visual deprivation, regardless of the integrity of the visual system, induces hypersensitivity. Moreover, the present findings indicate that late visual deprivation may induce mechanical and thermal hypersensitivity, although this depends on visual deprivation duration. These results have implications for the biological significance of pain in the blind. SIGNIFICANCE: Sensory deprivation induces behavioural adaptions. For most sensory systems, the extent of these adaptations generally depends on the stage of cerebral development. In contrast, the present results indicate that for the nociceptive system, both early and late visual deprivation have similar effects. Anophthalmic, dark-reared mice and adult mice deprived of vision for two months showed thermal and mechanical hypersensitivity. This shows a clear interaction between visual and nociceptive systems and has implications for the biological significance of pain in the blind.

Structural brain plasticity induced by early blindness.

It is well established that early blindness results in behavioural adaptations. While the functional effects of visual deprivation have been well researched, anatomical studies are scarce. The aim of this study was to investigate whole brain structural plasticity in a mouse model of congenital blindness. Volumetric analyses were conducted on high-resolution MRI images and histological sections from the same brains. These morphometric measurements were compared between anophthalmic and sighted ZRDBA mice obtained by breeding ZRDCT and DBA mice. Results from MRI analyses using the Multiple Automatically Generated Templates (MAGeT) method showed smaller volume for the primary visual cortex and superior colliculi in anophthalmic compared with sighted mice. Deformation-based morphometry revealed smaller volumes within the dorsal lateral geniculate nuclei and the lateral secondary visual cortex and larger volumes within olfactory areas, piriform cortex, orbital areas and the amygdala, in anophthalmic compared with sighted mice. Histological analyses revealed a larger volume for the amygdala and smaller volume for the superior colliculi, primary visual cortex and medial secondary visual cortex, in anophthalmic compared with sighted mice. The absence of superficial visual layers of the superior colliculus and the thinner cortical layer IV of the primary and secondary visual cortices may explain the smaller volume of these areas, although this was observed in a limited sample. The present study shows large-scale brain plasticity in a mouse model of congenital blindness. In addition, the congruence of MRI and histological findings support the use of MRI to investigate structural brain plasticity in the mouse.

Better Olfactory Performance and Larger Olfactory Bulbs in a Mouse Model of Congenital Blindness.

It is well established that early blindness results in enhancement of the remaining nonvisual sensory modalities accompanied by functional and anatomical brain plasticity. While auditory and tactile functions have been largely investigated, the results regarding olfactory functions remained less explored and less consistent. In the present study, we investigated olfactory function in blind mice using 3 tests: the buried food test, the olfactory threshold test, and the olfactory performance test. The results indicated better performance of blind mice in the buried food test and odor performance test while there was no difference in the olfactory threshold test. Using histological measurements, we also investigated if there was anatomical plasticity in the olfactory bulbs (OB), the most salient site for olfactory processing. The results indicated a larger volume of the OB driven by larger glomerular and granular layers in blind mice compared with sighted mice. Structural plasticity in the OB may underlie the enhanced olfactory performance in blind mice.

Pain Hypersensitivity is Associated with Increased Amygdala Volume and c-Fos Immunoreactivity in Anophthalmic Mice.

It is well established that early blindness results in brain plasticity and behavioral changes in both humans and animals. However, only a few studies have examined the effects of blindness on pain perception. In these studies, pain hypersensitivity was reported in early, but not late, blind humans. The underlying mechanisms remain unclear, but considering its key role in pain perception and modulation, the amygdala may contribute to this pain hypersensitivity. The first aim of this study was to develop an animal model of early blindness to examine the effects of blindness on pain perception. A mouse cross was therefore developed (ZRDBA mice), in which half of the animals are born sighted and half are born anophthalmic, allowing comparisons between blind and sighted mice with the same genetic background. The second aim of the present study was to examine mechanical and thermal pain thresholds as well as pain behaviors and pain-related c-Fos immunoreactivity induced by the formalin test in the amygdalas of blind and sighted mice. Group differences in amygdala volume were also assessed histologically. Blind mice exhibited lower mechanical and thermal pain thresholds and more pain behaviors during the acute phase of the formalin test, compared with sighted mice. Moreover, pain hypersensitivity during the formalin test was associated with increased c-Fos immunoreactivity in the amygdala. Furthermore, amygdala volume was larger bilaterally in blind compared with sighted mice. These results indicate that congenitally blind mice show pain hypersensitivity like early blind individuals and suggest that this is due in part to plasticity in the amygdala.

Regulation of cortical blood flow responses by the nucleus basalis of Meynert during nociceptive processing.

Cerebral blood flow (CBF) is essential for neuronal metabolic functions. CBF is partly regulated by cholinergic projections from the nucleus basalis of Meynert (NBM) during cortical processing of sensory information. During pain-related processing, however, this mechanism may be altered by large fluctuations in systemic mean arterial pressure (MAP). The objective of this study was to investigate the contribution of NBM to CBF responses evoked by nociceptive electrical stimuli and how it may be affected by systemic MAP. CBF was recorded in isoflurane-anesthetized rats (n = 8) using laser speckle contrast imaging, in two conditions (intact vs left NBM lesion). Electrical stimulation was applied to the sciatic nerve. Sciatic stimulation produced intensity dependent increases in MAP (p < 0.001) that were almost identical between conditions (intact vs left NBM lesion; p = 0.96). In both conditions, sciatic stimulation produced intensity dependent CBF increases (p < 0.001). After NBM lesion, CBF responses were decreased in the left somatosensory cortex ipsilateral to NBM lesion (p = 0.02) but not in the right somatosensory cortex (p = 0.46). These results indicate that NBM contributes to CBF responses to nociceptive stimulation in the ipsilateral, but not contralateral somatosensory cortex and that CBF response attenuation by NBM lesion is not compensated passively by systemic MAP changes. This highlights the importance of NBM's integrity for pain-related hemodynamic responses in the somatosensory cortex.

Sympathetic regulation and anterior cingulate cortex volume are altered in a rat model of chronic back pain.

Chronic pain is associated with autonomic disturbance. However, specific effects of chronic back pain on sympathetic regulation remain unknown. Chronic pain is also associated with structural changes in the anterior cingulate cortex (ACC), which may be linked to sympathetic dysregulation. The aim of this study was to determine whether sympathetic regulation and ACC surface and volume are affected in a rat model of chronic back pain, in which complete Freund Adjuvant (CFA) is injected in back muscles. Sympathetic regulation was assessed with renal blood flow (RBF) changes induced by electrical stimulation of a hind paw, while ACC structure was examined by measuring cortical surface and volume. RBF changes and ACC volume were compared between control rats and rats injected with CFA in back muscles segmental (T10) to renal sympathetic innervation or not (T2). In rats with CFA, chronic inflammation was observed in the affected muscles in addition to increased nuclear factor-kappa B (NF-kB) protein expression in corresponding spinal cord segments (p=0.01) as well as decreased ACC volume (p<0.05). In addition, intensity-dependent decreases in RBF during hind paw stimulation were attenuated by chronic pain at T2 (p's<0.05) and T10 (p's<0.05), but less so at T10 compared with T2 (p's<0.05). These results indicate that chronic back pain alters sympathetic functions through non-segmental mechanisms, possibly by altering descending regulatory pathways from ACC. Yet, segmental somato-sympathetic reflexes may compete with non-segmental processes depending on the back region affected by pain and according to the segmental organization of the sympathetic nervous system.

A three-dimensional digital atlas of the starling brain.

Because of their sophisticated vocal behaviour, their social nature, their high plasticity and their robustness, starlings have become an important model species that is widely used in studies of neuroethology of song production and perception. Since magnetic resonance imaging (MRI) represents an increasingly relevant tool for comparative neuroscience, a 3D MRI-based atlas of the starling brain becomes essential. Using multiple imaging protocols we delineated several sensory systems as well as the song control system. This starling brain atlas can easily be used to determine the stereotactic location of identified neural structures at any angle of the head. Additionally, the atlas is useful to find the optimal angle of sectioning for slice experiments, stereotactic injections and electrophysiological recordings. The starling brain atlas is freely available for the scientific community.