Lucy Stiles

The vestibular-basal ganglia connection: balancing motor control.

Connections between the vestibular system and the basal ganglia have been sporadically studied over the last century. Electrophysiological studies of field potentials in animals have shown that most areas of the striatum respond to electrical vestibular stimulation while human studies isolated responses to vestibular stimulation to the putamen of the striatum. Protein studies have shown inconsistent results regarding changes in receptor levels of a number of receptor types. Recent tracer studies identified a pathway between the vestibular nucleus and the striatum via the thalamus, completely bypassing the cortex. Vestibular sensory input is represented in the part of the striatum - the dorsolateral striatum - where fibres from the sensorimotor areas terminate. It is therefore possible that vestibular signals are used together with other sensorimotor inputs in the striatum for body and limb control. The combination of electrophysiological results, changes in protein levels and tracer studies have led to the idea that the dorsolateral striatum is likely to be the main input area for vestibular signals in the basal ganglia and these will have an influence on motor control. This may have clinical implications in the treatment of basal ganglia disorders and other movement disorders.

Acoustic trauma that can cause tinnitus impairs impulsive control but not performance accuracy in the 5-choice serial reaction time task in rats

Although tinnitus is an auditory disorder, it is often associated with attentional and emotional problems. Functional neuroimaging studies in humans have revealed that the hippocampus, amygdala and anterior cingulate, areas of the brain involved in emotion, attention and spatial processing, are also involved in auditory memory and tinnitus perception. However, few studies of tinnitus-evoked emotional and cognitive changes have been reported using animal models of tinnitus. In the present study, we investigated whether acoustic trauma that could cause tinnitus would affect attention and impulsivity in rats. Eight male Wistar rats were exposed to unilateral acoustic trauma (110 dB, 16 kHz for 1 h under anaesthesia) and eight rats underwent the same anaesthesia without acoustic trauma. Tinnitus was tested in noise-exposed rats using a frequency-specific shift in a discrimination function with a conditioned lick suppression paradigm. At 4 months after the noise exposure, the rats were tested in a 5-choice serial reaction time task. The behavioural procedure involved training the rats to discriminate a brief visual stimulus presented randomly in one of the five spatial locations and responding by poking its nose through the illuminated hole and collecting a food pellet from the magazine. While all of the animals performed equally well in making correct responses, the animals exposed to acoustic trauma made significantly more premature responses. The results suggest that rats exposed to acoustic trauma and some of which have chronic tinnitus are impaired in impulsive control, but not performance accuracy.

The D2 dopamine receptor and locomotor hyperactivity following bilateral vestibular deafferentation in the rat

Rats and mice with bilateral vestibular loss exhibit dramatic locomotor hyperactivity and circling behaviours, which to date cannot be explained. Dysfunction of the striatal dopaminergic system is responsible for a number of known movement disorders and the D(2) dopamine receptor is known to be implicated. Therefore, it is possible that changes in striatal function are responsible for locomotor hyperactivity and circling following bilateral vestibular lesions. The aim of this study was to investigate the effects of the D(2) receptor antagonist, eticlopride (0.02, 0.04 and 0.06mg/kg; s.c.), on locomotor behaviour in rats at 5 months following bilateral vestibular deafferentation (BVD), using an open field maze. The levels of the D(2) receptor protein in the striatum were measured at 1 and 6 months post-BVD using western blotting. BVD rats exhibited locomotor hyperactivity and circling, which eticlopride did not eliminate. However, BVD rats did exhibit a decreased response to the inhibitory effect of eticlopride compared to sham controls at the 0.02 mg/kg dose. There were no changes in the amount of the D(2) receptor in the striatum at 1 or 6 months post-BVD; however, D(2) receptor levels were significantly higher on the right side than the left in both sham and BVD animals. These results suggest that locomotor hyperactivity and circling behaviours following BVD are not due simply to changes in D(2) receptor protein expression in the striatum and that other neurophysiological changes in the brain account for these behaviours following BVD.

Evidence That Memantine Reduces Chronic Tinnitus Caused By Acoustic Trauma In Rats

Subjective tinnitus is a chronic neurological disorder in which phantom sounds are perceived. Increasing evidence suggests that tinnitus is caused by neuronal hyperactivity in auditory brain regions, either due to a decrease in synaptic inhibition or an increase in synaptic excitation. One drug investigated for the treatment of tinnitus has been the uncompetitive N-methyl-d-aspartate (NMDA) receptor antagonist, memantine, although the evidence relating to it has been unconvincing to date. We re-investigated the effects of memantine on the behavioral manifestations of tinnitus induced by acoustic trauma (a 16-kHz, 110-dB pure tone presented unilaterally for 1 h) in rats. We used a conditioned lick suppression model in which lick suppression was associated with the perception of high frequency sound resembling tinnitus and a suppression ratio (SR) was calculated by comparing the number of licks in the 15-s period preceding the stimulus presentation (A) and the 15-s period during the stimulus presentation (B), i.e., SR = B/(A + B). Acoustic trauma resulted in a significant increase in the auditory brainstem-evoked response (ABR) threshold in the affected ear (P ≤ 0.0001) and a decrease in the SR compared to sham controls in response to 32 kHz tones in five out of eight acoustic trauma-exposed animals. A 5-mg/kg dose of memantine significantly reduced the proportion of these animals which exhibited tinnitus-like behavior (2/5 compared to 5/5; P ≤ 0.006), suggesting that the drug reduced tinnitus. These results suggest that memantine may reduce tinnitus caused by acoustic trauma.

The effects of the synthetic cannabinoid receptor agonists, WIN55,212-2 and CP55,940, on salicylate-induced tinnitus in rats.

Previous studies in animals and humans have shown that, in some cases at least, anti-epileptic drugs can reduce the severity of tinnitus. Given that cannabinoid receptor agonists have been shown to exert anti-epileptic effects in some circumstances, we investigated whether two synthetic CB(1)/CB(2) receptor agonists, WIN55,212-2, and CP55,940, could inhibit the behavioural manifestations of salicylate-induced tinnitus in rats in a conditioned suppression task. We found that neither WIN55,212-2 (3.0 mg/kg s.c) nor CP55,940 (0.1 or 0.3 mg/kg s.c), significantly reduced conditioned behaviour associated with tinnitus. However, both 3 mg/kg WIN55,212-2 and 0.3 mg/kg CP55,940 did significantly increase tinnitus-related behaviour compared to the vehicle control groups. These results suggest that cannabinoid receptor agonists may not be useful in the treatment of salicylate-induced tinnitus and that at certain doses, they could actually exacerbate the condition.

Galvanic vestibular stimulation impairs cell proliferation and neurogenesis in the rat hippocampus but not spatial memory

Galvanic vestibular stimulation (GVS) is a method of activating the peripheral vestibular system using direct current that is widely employed in clinical neurological testing. Since movement is recognized to stimulate hippocampal neurogenesis and movement is impossible without activation of the vestibular system, we speculated that activating the vestibular system in rats while minimizing movement, by delivering GVS under anesthesia, would affect hippocampal cell proliferation and neurogenesis, and spatial memory. Compared with the sham control group, the number of cells incorporating the DNA replication marker, bromodeoxyuridine (BrdU), was significantly reduced in the bilateral hippocampi in both the cathode left‐anode right and cathode right‐anode left stimulation groups (P ≤ 0.0001). The majority of the BrdU+ve cells co‐expressed Ki‐67, a marker for the S phase of the cell cycle, suggesting that these BrdU+ve cells were still in the cell cycle; however, there was no significant difference in the degree of co‐labeling between the two stimulation groups. Single labeling for doublecortin (DCX), a marker of immature neurons, showed that while there was no significant difference between the different groups in the number of DCX+ve cells in the right dentate gryus, in the left dentate gyrus there was a significant decrease in the cathode left‐anode right group compared with the sham controls (P ≤ 0.03). Nonetheless, when animals were tested in place recognition, object exploration and Morris water maze tasks, there were no significant differences between the GVS groups and the sham controls. These results suggest that GVS can have striking effects on cell proliferation and possibly neurogenesis in the hippocampus, without affecting spatial memory.

Glutamate Receptor Subunit and Calmodulin Kinase II Expression, with and without T Maze Training, in the Rat Hippocampus following Bilateral Vestibular Deafferentation

Many previous studies have shown that lesions of the peripheral vestibular system result in spatial memory deficits and electrophysiological dysfunction in the hippocampus. Given the importance of glutamate as a neurotransmitter in the hippocampus, it was predicted that bilateral vestibular deafferentation (BVD) would alter the expression of NMDA and AMPA receptors in this area of the brain. Methods The expression of the NR1, NR2A, NR2B, GluR1, GluR2, GluR3 and GluR 4 glutamate receptor subunits, as well as calmodulin kinase IIα (CaMKIIα) and phosphorylated CaMKIIα (pCaMKIIα), was measured in the rat CA1, CA2/3 and dentate gyrus (DG) subregions of the hippocampus, at 24 h, 72 h, 1 week, 1 month and 6 months following BVD, using western blotting. In the 6 month group, half of the animals underwent spatial forced alternating training in a T-maze. Results and Discussion For the 24 h, 72 h, 1 week and 1 month data, there was no significant effect of surgery for any hippocampal subregion. However, for the 6 month data set, T maze training had a significant effect independently of surgery. The results of these experiments suggest that BVD is not associated with large changes in glutamate receptor subunit or CaMKIIα expression in the rat hippocampus, at least in terms of both the intra-cytoplasmic and membrane receptor subunits together, that western blotting can measure. However, spatial training-associated increases in glutamate receptor and CaMKIIα expression can be induced in BVD rats with impaired spatial performance. Therefore, the neurophysiological changes underlying BVD-induced spatial learning and memory deficits are more likely to be due to up and down regulation or changes in affinity/efficacy of glutamate receptors at the membrane level than changes in subunit transcription and transduction at the intra-cytoplasmic level.

Cell proliferation in the cochlear nucleus following acoustic trauma in rat

Our previous studies have suggested that surgical lesions of the rat cochlea induce cell proliferation in the cochlear nucleus (CN) that may be related to neurogenesis. The aim of the present study was to further investigate the nature of cell proliferation in the CN, following acoustic trauma that has previously been shown to induce tinnitus in rats. Rats were subjected either to a unilateral acoustic trauma (16-kHz pure tone, 115dB for 1h under anesthesia) or a sham procedure. Bromodeoxyuridine (BrdU) immunohistochemistry was used to measure cell proliferation and newborn cell survival; an antibody to interleukin-6 was used to investigate inflammatory responses; and double immunolabeling for BrdU and Ki-67, BrdU and CD-11b, and BrdU and doublecortin (DCX), was used to investigate the origin of the proliferating cells. There was a time-dependent increase in the number of BrdU(+ve) cells in the CN following acoustic trauma; however, the number of BrdU(+ve) cells that survived was comparable to that of control animals at 4 weeks post-trauma. Cell proliferation was unlikely to be due to proliferating inflammatory cells as a result of a trauma-induced inflammatory response as the IL-6 expression level was comparable between sham and exposed groups. Immunolabeling revealed the BrdU(+ve) cells to co-express Ki-67 and DCX, but not CD-11b. However, there was no difference in DCX expression between sham and exposed animals. The results suggest that DCX-expressing cells in the CN may proliferate in response to acoustic trauma; however, the proportion of cells proliferating and the survival rate of the newborn cells may not support functional neurogenesis in the CN.

The Effects of Acute Stress-Induced Sleep Disturbance on Acoustic Trauma-Induced Tinnitus in Rats

Chronic tinnitus is a debilitating condition and often accompanied by anxiety, depression, and sleep disturbance. It has been suggested that sleep disturbance, such as insomnia, may be a risk factor/predictor for tinnitus-related distress and the two conditions may share common neurobiological mechanisms. This study investigated whether acute stress-induced sleep disturbance could increase the susceptibility to acoustic trauma-induced tinnitus in rats. The animals were exposed to unilateral acoustic trauma 24 h before sleep disturbance being induced using the cage exchange method. Tinnitus perception was assessed behaviourally using a conditioned lick suppression paradigm 3 weeks after the acoustic trauma. Changes in the orexin system in the hypothalamus, which plays an important role in maintaining long-lasting arousal, were also examined using immunohistochemistry. Cage exchange resulted in a significant reduction in the number of sleep episodes and acoustic trauma-induced tinnitus with acoustic features similar to a 32 kHz tone at 100 dB. However, sleep disturbance did not exacerbate the perception of tinnitus in rats. Neither tinnitus alone nor tinnitus plus sleep disturbance altered the number of orexin-expressing neurons. The results suggest that acute sleep disturbance does not cause long-term changes in the number of orexin neurons and does not change the perception of tinnitus induced by acoustic trauma in rats.

Effects of acute altered gravity during parabolic flight and/or vestibular loss on cell proliferation in the rat dentate gyrus

Both parabolic flight, i.e. a condition of altered gravity, and loss of vestibular function, have been suggested to affect spatial learning and memory, which is known to be influenced by neurogenesis in the hippocampus. In this study we investigated whether short alternated micro- and hyper-gravity stimulations during parabolic flight and/or loss of vestibular function, would alter cell proliferation in the hippocampal dentate gyrus of rats, by measuring the number of bromodeoxyuridine (BrdU)-incorporated cells. Rats were randomly allocated to the following experimental groups: (1) sham transtympanic saline injection only (n=5); (2) bilateral vestibular deafferentation (BVD) by sodium arsanilate transtympanic injection only (n=5); (3) sham treatment and parabolic flight (n=5); (4) BVD and parabolic flight (n=6). Forty-two days following transtympanic injection, the animals were subjected to parabolic flight in an awake restrained condition after habituation. A modified Airbus A300 aircraft was flown on a parabolic path, creating 20s of 1.8G during both climbing and descending and 22s of 0G at the apex of each parabola. The no flight animals were subjected to the same housing for the same duration. Immediately after the parabolic flight or control ground condition, animals were injected with BrdU (300mg/kg, i.p). Twenty-four hs after BrdU injection, rats were sacrificed. BrdU immunolabelling was performed and the number of BrdU+ve cells in the dentate gyrus of the hippocampus was quantified using a modified fractionator method. BVD caused a large and significant reduction in the number of BrdU-positive cells compared to sham animals (P≤0.0001); however, flight and all interactions were non-significant. These results indicate that BVD significantly decreased cell proliferation irrespective of the short exposure to altered/modified gravity.