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 chronic tinnitus caused by acoustic trauma on social behaviour and anxiety in rats.

Tinnitus is associated with significant increases in anxiety disorders in humans, which is thought to affect social interaction; however, there has been only one previous study of the effects of tinnitus on social interaction in animals treated with salicylate and no previous study of the effects of tinnitus on anxiety in animals. In the present study, we used acoustic trauma to induce tinnitus in rats and investigated its effects on social interaction and anxiety in animals confirmed to have tinnitus. When social behaviours were grouped, we found that animals with tinnitus engaged in significantly more aggressive behaviours toward both tinnitus and sham control animals (P=0.03). When individual social behaviours were analysed without considering whether a tinnitus or sham animal was interacting with a member of its own treatment group, tinnitus animals were found to engage in significantly more anogenital investigation (P=0.01) and significantly less social grooming (P=0.003). When the data were analysed according to whether an animal was interacting with a member of its own group, tinnitus animals were found to bite sham animals significantly more than other tinnitus animals (P=0.005). Sham animals also bit tinnitus animals significantly more than other sham animals (P=0.02), as well as climbing away from them more (P=0.04), kicking (P=0.003), nudging them more (P=0.04), and sleeping with them more (P=0.02). By contrast, sham animals sniffed tinnitus animals significantly less than sham animals (P=0.05). There were no significant differences between the sham and tinnitus animals in performance in the elevated plus and elevated T maze tests of anxiety. However, tinnitus animals displayed a slight but significant increase in locomotor activity in the open field (P=0.04). These data suggest that tinnitus results in complex changes in social interaction in rats, which are not due simply to increases in anxiety.

The effects of acoustic trauma that can cause tinnitus on spatial performance in rats.

Previous studies have shown that acoustic trauma can disrupt the firing of place cells in the hippocampus and also inhibit hippocampal neurogenesis, suggesting that such trauma might impair spatial memory. In this study, we investigated performance in the alternating T maze and Morris water maze of rats exposed to acoustic trauma (16 kHz, 110 dB SPL pure tone for 1 h), who had elevated auditory brainstem response thresholds and the psychophysical attributes of tinnitus (using a conditioned lick suppression task). To our surprise, we found that rats with tinnitus did not perform significantly differently from sham control animals in either the alternating T maze task or any aspect of the reference or working memory versions of the Morris water maze task except for a faster acquisition in T maze alternation. These results suggest that acoustic trauma and tinnitus may not impair spatial memory in rats.

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.

Potential involvement of NOS and arginase in age-related behavioural impairments.

The present study investigated age-related changes in nitric oxide synthase (NOS) and arginase, which shares a substrate with NOS, in the hippocampus and parahippocampal region and the relationship between NOS/arginase and age-associated behavioural impairments. Aged rats (24 months old) displayed reduced exploratory activity, enhanced anxiety, poorer spatial learning and memory, and impaired object recognition memory relative to the young adults (4 months old). There were significant increases in total NOS activity in the aged hippocampus and perirhinal, postrhinal and temporal cortices and a dramatic decrease in endothelial NOS expression in the aged postrhinal cortex. Activity and protein expression of inducible NOS were not detected in any region from either group and a significant increase in total arginase activity was found in the aged perirhinal cortex. Multiple regression analysis revealed significant correlations between NOS/arginase and behavioural measures in both groups. The present findings provide further support for a contribution of nitric oxide to the normal aging process and suggest a potential involvement of arginase in aging and learning and memory.

Age-related changes in nitric oxide synthase and arginase in the rat prefrontal cortex.

Increasing evidence suggests that nitric oxide (NO), generated by nitric oxide synthase (NOS) from l-arginine, plays an important role in the ageing process. The present study, for the first time, investigates age-related changes in NOS and arginase, an enzyme that shares a common substrate with NOS, in the prefrontal cortex of rats assessed with and without prior behavioural testing. A significant increase in total NOS activity was found in the prefrontal cortex in aged (24-month-old) as compared with young (4-month-old) rats. Western blotting revealed that there were no significant differences between young and aged rats in neuronal NOS (nNOS) and endothelial NOS (eNOS) protein expression. Inducible isoform of NOS (iNOS), in terms of activity and protein expression, was not detected in either group. Total arginase activity and arginase I and II protein expression did not differ between the young and aged groups. The present findings support the contribution of NOS/NO to ageing but question the importance of iNOS in the normal ageing process.

Regional variations and age-related changes in nitric oxide synthase and arginase in the sub-regions of the hippocampus.

L-arginine can be metabolised by nitric oxide synthase (NOS) with the formation of L-citrulline and nitric oxide (NO), or arginase with the production of L-ornithine and urea. In contrast to studies showing a potential involvement of NOS/NO in the aging process, the role of arginase has not been well documented. The present study investigates for the first time the regional variations and age-related changes in both NOS and arginase in sub-regions of the hippocampus. In young adult rats, although the total NOS activity was not significantly different across the hippocampal CA1, CA2/3 and the dentate gyrus (DG) sub-regions, the total arginase activity showed a clear regional variation with the highest level in DG. Western blotting revealed that the highest levels of neuronal NOS (nNOS) and endothelial NOS (eNOS) proteins were located in CA1. Arginase I is expressed at a very low level in the brain (the whole hippocampus) as compared with the liver. By contrast, arginase II protein shows an extremely high expression in the brain with little or no expression in the liver. There was no regional variation in arginase I or arginase II protein expression across the sub-regions of the hippocampus. When a comparison was made between young (4-month-old) and aged (24-month-old) rats, a significant increase in total NOS activity was found in DG and significant decreases in arginase activity were observed in the CA1 and CA2/3 regions in the aged animals. Western blotting further revealed a dramatic decrease in eNOS protein expression in aged CA2/3 with no age-associated changes in nNOS, arginase I and II protein expression in any region examined. Interestingly, evidence of activity or protein expression of the inducible isoform of NOS (iNOS) was not detected in any tissue from either group. The present results, in conjunction with previous findings, support the contribution of NOS/NO to aging but question the involvement of iNOS in the normal aging process. Region-specific changes in arginase suggest that this enzyme may also contribute to aging.

Long-term changes in hippocampal n-methyl-D-aspartate receptor subunits following unilateral vestibular damage in rat.

Previous studies have indicated that damage to the peripheral vestibular system results in dysfunction of hippocampal place cells and an impairment of spatial learning and memory. The aim of this study was to determine whether lesions of one vestibular labyrinth (unilateral vestibular deafferentation, UVD) result in changes in the expression of the NR1 and NR2A subunits of the N-methyl-D-aspartate (NMDA) receptor, and the GluR2 subunit of the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptor, in subregions of the rat hippocampus (CA1, CA2/3 and the dentate gyrus) at 10 h or 2 weeks following UVD. Compared with sham surgery controls and anaesthetic controls, the expression of the NR1 subunit was significantly reduced in the ipsilateral CA2/3 region at 2 weeks post-UVD. The expression of the NR2A subunit was also significantly reduced in the ipsilateral CA2/3 and, to a smaller extent, in the contralateral CA2/3 region, at 2 weeks post-UVD. The only other change in NR2A expression was an increase in the ipsilateral CA1 at 10 h post-UVD. No other changes in NR1, NR2A or GluR2 expression were observed in any hippocampal subregion, at any time point, or in cortical tissue at any time point. These results suggest that UVD may result in long-term changes in NMDA receptor subunit expression in the rat hippocampus.

The contribution of nitric oxide to vestibular compensation: are there species differences?

Nitric oxide (NO) has been implicated in the processes by which animals recover from peripheral vestibular damage ("vestibular compensation"). However, there is little systematic data available on the effects of NO inhibition on the vestibular compensation process. In the present study we administered the nitric oxide synthase (NOS) inhibitor NG-nitro-L-arginine methyl ester (L-NAME) using a subcutaneous osmotic minipump and examined its effects on the compensation of spontaneous nystagmus (SN), yaw head tilt (YHT) and roll head tilt (RHT) in guinea pigs. Following unilateral labyrinthectomy (UL), treatment with 5, 10, 50 or 100 mM L-NAME had no effect on the expression of any of these symptoms or their rate of compensation. By contrast, pre-UL treatment with 100 mM L-NAME resulted in a decrease in SN frequency at 10 h post-UL and an increase in its rate of compensation. Lower concentrations had no effect on SN. Pre-UL treatment with L-NAME had no significant effect on YHT or RHT at any particular time point. Analysis of NOS activity demonstrated that the highest concentration of L-NAME inhibited NOS activity in the contralateral vestibular nucleus complex, bilateral cerebellum and bilateral cortices. These results suggest that L-NAME may have different effects on vestibular compensation in guinea pigs compared to other species, such as the rat and frog.

Quantitative changes in gene expression of glutamate receptor subunits/subtypes in the vestibular nucleus, inferior olive and flocculus before and following unilateral labyrinthectomy in the rat: real-time quantitative PCR method.

Spontaneous recovery from the oculomotor and postural symptoms of unilateral labyrinthectomy (UL) is known as vestibular compensation, which is a useful model for investigation of the mechanisms of lesion-induced CNS plasticity. In the present study, to elucidate the molecular biological basis of vestibular compensation, we investigated changes in the mRNA expression of glutamate receptor subunit/subtypes in the rat central vestibular system, including the vestibular nucleus complex (VNC), inferior olive (IO), and cerebellar flocculus following UL, using a real-time quantitative polymerase chain reaction (PCR) method. In normal control animals, regional differences in the expression of several glutamate receptor subunit/subtypes, e.g., NR1 and NR2A subunits of the N-methyl-D-aspartic acid (NMDA) receptor, GluR2 and KA2 subtypes of non-NMDA receptors, and mGluR1 and mGluR7 metabotropic glutamate receptors, were consistent with previous results from studies using in situ hybridization histochemistry, suggesting that the real-time quantitative PCR method was a reliable procedure for evaluation of changes in mRNA expression. In the vestibular nucleus complex, NR2A, GluR2 and mGluR7 mRNA were ipsilaterally downregulated by 6 h following UL (P<0.05, P<0.05 and P<0.01, respectively). In the inferior olive, no changes in gene expression were observed. In the ipsilateral flocculus, KA2 mRNA expression was increased by 50 h post-UL (P<0.05). However, in the contralateral flocculus, mGluR1 mRNA was downregulated by 6 h post-UL (P<0.005). Both the increase in KA2 mRNA expression in the ipsilateral flocculus and the decrease in mGluR1 mRNA expression in the contralateral flocculus may have had the effect of reducing Purkinje cell inhibition of ipsilateral VNC neurons, thereby contributing to the rebalancing of spontaneous resting activity between the ipsilateral and contralateral VNCs. It is suggested that such changes in the activities of the floccular-VNC pathways may be important to the vestibular compensation process.

Damage to the vestibular inner ear causes long-term changes in neuronal nitric oxide synthase expression in the rat hippocampus.

The vestibular inner ear detects head acceleration and initiates compensatory eye movement and postural reflexes that help keep the visual image of the world stable on the retina, and maintain balance, during unexpected head movement. The most primitive vestibular systems are estimated to have evolved more than 500 million years ago and in mammalian and submammalian species the vestibular reflexes are mediated by basic brainstem pathways (see Wilson and Melvill Jones, 1979 for review). Although the contributions of the vestibular system to higher cognitive function have generally received less attention than its reflexive roles, vestibular sensory information is transmitted to higher centres in the brain and humans with vestibular damage are known to experience debilitating perceptual illusions (see Curthoys and Halmagyi, 1995; Berthoz, 1996 for reviews). Increasing behavioural and neurophysiological evidence suggests that the hippocampus uses information from the vestibular inner ear in order to build up maps of space that can be used in the development of spatial memory during learning tasks (McNaughton et al., 1991; Chapuis et al., 1992; Wiener and Berthoz, 1993; O'Mara et al., 1994; Wiener et al., 1995; Gavrilov et al., 1995; Stackman and Taube, 1996; Vitte et al., 1996; Taube et al., 1996; Save et al., 1998; Peruch et al., 1999; Cuthbert et al., 2000; Russell et al., 2000). However, to date, there has been no indication of the long-term neurochemical effects of the loss of vestibular input on hippocampal function. Since nitric oxide has been implicated in the mechanisms of hippocampal synaptic plasticity associated with the development of short-term memory (e.g. Schuman and Madison, 1991; Schuman et al., 1994; Arancio et al., 1996; Wu et al., 1997; Lu et al., 1999), we examined whether changes occur in the activity and expression of the enzymes responsible for nitric oxide production (nitric oxide synthases) in subregions of the rat hippocampus at different times following unilateral peripheral vestibular lesions, using western blotting and radioenzymatic assays. We found a decreased expression of neuronal nitric oxide synthase in the ipsilateral dentate gyrus at 2 weeks following the vestibular damage and not before, that may be related to the long-term effects of the loss of vestibular input on hippocampal function. These results support the hypothesis that head movement and position information derived from the vestibular inner ear may be important for the normal function of the hippocampus.

Menstrual cycle effects on postural stability but not optokinetic function.

We investigated the effects of the menstrual cycle on visual-vestibular interaction by measuring optokinetic function and postural stability at different phases of the cycle. Menstrual cycle phase had no significant effect on gaze-holding, optokinetic nystagmus slow phase velocity, amplitude or frequency, circularvection latency or optokinetic afternystagmus frequency, amplitude or duration. While menstrual cycle phase had no significant effect on anterior-posterior sway, it did significantly affect lateral sway (P<0.001), with sway on day 5 significantly greater than on days 12 and 21 (P<0.05 and 0.01, respectively), and sway on day 25 significantly greater than that on day 21 (P<0.05).

Optokinetic reflex dysfunction in multiple sclerosis.

The aim of this study was to further investigate optokinetic reflex function in multiple sclerosis. Gaze-holding in darkness, optokinetic nystagmus, optokinetic afternystagmus and latency to circularvection were measured using electro-oculography and a rotating optokinetic drum. Gaze-holding was not significantly different between the multiple sclerosis and control groups; however, four of 23 multiple sclerosis patients exhibited eccentric gaze-evoked nystagmus. There were no significant differences in either optokinetic nystagmus frequency or latency to circularvection. However, optokinetic nystagmus slow phase velocity during rise time and amplitude during beat time were significantly reduced in the multiple sclerosis group (p < 0.05 and p < 0.0001, respectively). The time constant of optokinetic afternystagmus was also significantly reduced in the multiple sclerosis group (p < 0.005). These results indicate that optokinetic nystagmus and optokinetic afternystagmus are significantly impaired multiple sclerosis.

Differences in NOS protein expression and activity in the rat vestibular nucleus following unilateral labyrinthectomy.

We used Western blotting to analyse the expression of different isoforms of nitric oxide synthase (NOS) in the rat vestibular nucleus complex (VNC) at various times following unilateral vestibular deafferentation (UVD), together with a radioenzymatic assay to compare NOS activity at the same time points. nNOS expression did not change significantly in the ipsilateral or contralateral VNC at any time following UVD. However, eNOS expression decreased significantly (P<0.05) in the contralateral VNC at 6 h post-UVD, recovering to normal levels by 50 h. iNOS was not expressed at any time following UVD. NOS activity demonstrated a significant increase in the contralateral VNC at 6 h post-UVD (P<0.05), recovering toward normal levels by 50 h.

The effects of protein kinase C and calmodulin kinase II inhibitors on vestibular compensation in the guinea pig.

Previous studies have demonstrated that vestibular compensation, the process of behavioural recovery which occurs following unilateral deafferentation of the vestibular labyrinth (UVD), is correlated with changes in in vitro phosphorylation of various protein substrates in the brainstem vestibular nucleus complex (VNC). The aim of the present study was to investigate the possible causal relationship between protein kinase activity and the induction of the vestibular compensation process, by delivering inhibitors of protein kinase C (PKC) or Ca(2+)/calmodulin-dependent kinase II (CaMKII) into the ipsilateral VNC at the time of the UVD and determining their effects on three static symptoms of UVD, spontaneous nystagmus (SN), yaw head tilt (YHT) and roll head tilt (RHT) in guinea pigs. Infusion of the PKC inhibitor, 3-[1-(3-dimethylaminopropyl)-1H-indol-3-yl]-4-(1H-indol-3-yl)-1H-pyrr ole-2,5-dione, HCl (bisindolylmaleimide I, HCl/GF 109203X, HCl) ('Bis I'), at a concentration of 5 or 50 microM, significantly increased SN frequency at the earliest time points (6 and 8 h post-UVD) compared to vehicle controls and the less selective analogue, 2,3-bis(1H-indol-3-yl)-N-methylmaleimide (bisindolylmaleimide V) ('Bis V'). However, the compensation of YHT and RHT was unaffected by the PKC inhibitor. By contrast, the cell-permeable CaMKII inhibitor, myristoylated autocamtide-2 related inhibitory peptide (N-Myr-Lys-Lys-Ala-Leu-Arg-Arg-Gln-Glu-Ala-Val-Asp-Ala-Leu-OH) ('myr-AIP') or the cell-impermeable analogue, autocamtide-2 related inhibitory peptide (N-Lys-Lys-Ala-Leu-Arg-Arg-Cln-Glu-Ala-Val-Asp-Ala-Leu-OH) ('AIP'), failed to alter the compensation of SN, YHT or RHT at any dose compared to vehicle controls. These results implicate PKC-, but not CaMKII-, signal transduction pathways in the initiation of SN compensation in guinea pig.

The effects of repeated optokinetic stimulation on human autonomic function.

Numerous animal studies have suggested that the vestibular system modulates respiratory and cardiovascular function. However, relatively few studies have examined vestibular-autonomic interaction in humans. In this study we investigated the effects of repeated horizontal (clockwise or anticlockwise) optokinetic stimulation on systolic blood pressure (SBP), diastolic blood pressure (DBP), pulse and latency to circularvection (CV) in humans, and compared the effects with those of repeated exposure to a stationary visual stimulus. Although all subjects experienced CV, neither mean SBP, DBP or pulse differed significantly between the clockwise/anticlockwise optokinetic stimulation and no optokinetic stimulation conditions. However, SBP and DBP changed significantly over the 20 trials in each test session, even when there was no optokinetic stimulation (P < 0.001 in each case). These results suggest that while horizontal optokinetic stimulation does not significantly affect SBP, DBP or pulse in humans, changes in these variables can be induced by trial repetition itself, even when no optokinetic stimulation occurs.

Effects of intra-vestibular nucleus injection of the group I metabotropic glutamate receptor antagonist AIDA on vestibular compensation in guinea pigs.

Removal of the peripheral vestibular receptor cells in one inner ear (unilateral vestibular deafferentation, UVD) results in a syndrome of ocular motor and postural disorders, many of which disappear over time in a process of behavioural recovery known as vestibular compensation. Excitatory amino acid receptors, in particular the N-methyl-D-aspartate (NMDA) receptor, have been implicated in vestibular compensation; however, the metabotropic glutamate receptors (mGluRs) have not been studied in this context. The aim of this study was to determine whether group I mGluRs in the brainstem vestibular nucleus complex (VNC) ipsilateral to the UVD are involved in vestibular compensation of the static symptoms of UVD in guinea pig. The selective group I mGluR antagonist (RS)-1-aminoindan-1,5,dicarboxylic acid (AIDA) was continuously infused into the ipsilateral VNC for 30-min pre-UVD and 30-min post-UVD by cannula, at a rate of 1 microl/h, using one of four doses: 0.1 fg, 0.1 pg, 0.1 ng or 0.1 microg (n=5 animals in each case). In control conditions, a 0.1-fg (n=4) or 0.1-microg (n=5) NaOH vehicle was infused into the ipsilateral VNC using the same protocol. In order to control for the possibility that AIDA disrupted spontaneous neuronal activity in the VNC in normal animals, 0.1 microg AIDA (n=4) or 0.1 microg NaOH (n=2) was infused into the VNC in labyrinthine-intact animals. In both groups, static symptoms of UVD (i.e. spontaneous nystagmus, SN, yaw head tilt, YHT and roll head tilt, RHT) were measured at 8, 10, 12, 15, 20, 25, 30, 35, 45 and 50 h post-UVD. In addition, the righting reflex latency (RRL) was measured in labyrinthine-intact animals in order to assess whether AIDA impaired motor coordination in labyrinthine-intact animals. In UVD animals, the highest dose of AIDA significantly reduced SN frequency and changed its rate of compensation (P<0.001 and P<0.0001, respectively). This dose of AIDA also caused a significant reduction in YHT (P<0.005) as well as a significant change in its rate of compensation (P<0.0001). However, RHT was not significantly affected. In the labyrinthine-intact animals, AIDA infusion did not induce a UVD syndrome, nor did it significantly affect RRL. These results suggest that group I mGluRs in the ipsilateral VNC may be involved in the expression of ocular motor and some postural symptoms following UVD. Furthermore, group I mGluRs may not contribute to the resting activity of vestibular nucleus neurons.

The effects of L-NAME on vestibular compensation and NOS activity in the vestibular nucleus, cerebellum and cortex of the guinea pig.

Nitric oxide (NO) has been implicated in the processes by which animals recover from peripheral vestibular damage ('vestibular compensation'). However, few data exist on the dose-response effects of systemic administration of the nitric oxide synthase (NOS) inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME), on the vestibular compensation process. The aim of this study was to investigate the effects on compensation of 5, 10, 50 or 100 mM L-NAME administered by s.c osmotic minipump for 50 h following unilateral vestibular deafferentation (UVD) in guinea pig, either commencing the drug treatment at 4 h pre-UVD or at the time of the UVD (i.e., post-UVD). Post-UVD treatment with L-NAME, at any of the four concentrations used, had no effect on the compensation of spontaneous nystagmus (SN), yaw head tilt (YHT) or roll head tilt (RHT). By contrast, pre-UVD treatment with 100 mM L-NAME resulted in a significant decrease in SN frequency (P<0.05) and a change in the rate of its compensation (P<0.0005). Pre-UVD L-NAME resulted in a significant increase in the overall magnitude of YHT (P<0.005); however, post-hoc comparisons revealed no significant differences between any specific L-NAME and vehicle groups. Pre-UVD L-NAME had no effect on RHT at any concentration. Analysis of NOS activity in the pre-UVD L-NAME treatment groups at 50 h post-UVD showed that only 100 mM L-NAME resulted in a significant decrease in NOS activity in the contralateral medial vestibular nucleus (MVN)/prepositus hypoglossi (PH) (P<0.05) and that NOS activity in the ipsilateral MVN/PH was not significantly affected. However, NOS activity was significantly inhibited in the bilateral cerebellum and cortices for several concentrations of L-NAME. These results suggest that pre-UVD systemic administration of L-NAME can significantly increase the rate of SN compensation in guinea pig and that this effect is correlated with inhibition of NOS activity in several regions of the CNS.

The effects of scopolamine and cyclizine on visual-vestibular interaction in humans.

The aim of the present study was to investigate the effects of scopolamine (1.5 mg, transdermal patch) and cyclizine (50 mg tablet), at the doses usually used for the relief of motion sickness, on postural sway, optokinetic nystagmus (OKN) and circularvection (CV) in humans, using a within-subjects, double-blind, placebo-controlled design. Scopolamine and cyclizine were found to have no significant suppressive effect on these aspects of visual-vestibular interaction. Postural sway and CV were not significantly affected by either drug treatment; OKN SPV was significantly increased (p < 0.05), although OKN amplitude and frequency were unaffected. These results suggest that scopolamine and cyclizine, at doses used for the relief of motion sickness, may have minimal suppressive effects on these aspects of visual-vestibular interaction.

Comparison of protein kinase activity and protein phosphorylation in the medial vestibular nucleus and prepositus hypoglossi in labyrinthine-intact and labyrinthectomized guinea pigs.

The aim of the present study was to compare in vitro protein expression, protein kinase activity and protein phosphorylation in the medial vestibular nucleus (MVN) and prepositus hypoglossi (PH) from labyrinthine-intact guinea pigs and from guinea pigs at various stages of vestibular compensation following unilateral labyrinthectomy (UL). The ipsilateral (I-MVN) and contralateral (C-MVN) MVN, and the ipsilateral (I-PH) and contralateral (C-PH) PH, were dissected from 3 naive labyrinthine-intact guinea pigs and 55 guinea pigs at 10 hs or 53 hs following a surgical UL or sham operation. Tissue extracts were incubated with [gamma-33P]ATP+/-Ca2+, phorbol 12, 13 dibutyrate and phosphatidylserine or +/- Ca2+ and calmodulin, to enhance protein kinase C (PKC) or calcium calmodulin kinase (CaMK) activity, respectively. Data were analysed as the ratio of activated to basal 33P incorporation detected by phosphorimaging. There were similar total protein and phosphoprotein profiles in the MVN and PH, as well as both PKC and CaMKII activity, suggesting that the MVN and PH are similar in the way that proteins undergo rapid modification by phosphorylation. During the development of vestibular compensation, a 46 kDa band in C-PH displayed higher PKC-mediated phosphorylation from 10 hs post-UL compared to sham controls. Significantly greater PKC-mediated phosphorylation of proteins of approximately 18, 46 and 75 kDa was observed in C-PH at 10 hs compared to 53 hs post-UL and in most cases the phosphorylation was greater in C-PH than in the C-MVN. These results suggest that between 10 and 53 hs post-UL, PKC-mediated phosphorylation changes mainly in the C-PH rather than the ipsilateral or contralateral MVN.