TTX Injection Research Articles

Effect of Tetrodotoxin Pellets in a Rat Model of Postherpetic Neuralgia.

Postherpetic neuralgia (PHN) is nerve pain caused by a reactivation of the varicella zoster virus. Medications are used to reduce PHN but their use is limited by serious side effects. Tetrodotoxin (TTX) is a latent neurotoxin that can block neuropathic pain, but its therapeutic index is only 3–5 times with intravenous or intramuscular injection. Therefore, we prepared oral TTX pellets and examined their effect in a rat model of PHN induced by resiniferatoxin (RTX). Oral TTX pellets were significantly effective at preventing RTX-induced mechanical and thermal allodynia, and similar to pregabalin. Moreover, oral administration of TTX pellets dose-dependently inhibited RTX-induced PHN compared with intramuscular administration of TTX injection. We also studied the pharmacokinetic profile of TTX pellets. Our results showed that the blood concentration of TTX reached a maximum plasma concentration (Cmax) at around 2 h, with an elimination half-life time (t1/2) of 3.23 ± 1.74 h after intragastric administration. The median lethal dose (LD50) of TTX pellets was 517.43 μg/kg via oral administration to rats, while the median effective dose (ED50) was approximately 5.85 μg/kg, and the therapeutic index was 88.45. Altogether, this has indicated that oral TTX pellets greatly enhance safety when compared with TTX injection.

Time-Dependent Decline in Multifocal Electroretinogram Requires Faster Recording Procedures in Anesthetized Pigs.

PurposeThe time-dependent effect of anesthetics on the retinal function is debated. We hypothesize that in anesthetized animals there is a time-dependent decline that requires optimized multifocal electroretinogram (mfERG) recording procedures.MethodsConventional and four-frame global-flash mfERG recordings were obtained approximately 15, 60, and 150 minutes after the induction of propofol anesthesia (20 pigs) and isoflurane anesthesia (nine pigs). In six of the propofol-anesthetized pigs, the mfERG recordings were split in 3-minute segments. Two to 4 weeks after initial recordings, an intraocular injection of tetrodotoxin (TTX) was given and the mfERG was rerecorded as described above. Data were analyzed using mixed models in SAS statistical software.ResultsPropofol significantly decreases the conventional and global-flash amplitudes over time. The only significant effect of isoflurane is a decrease in the global-flash amplitudes. At 15 minutes after TTX injection several of the mfERG amplitudes are significantly decreased. There is a linear correlation between the conventional P1 and the global-flash DR mfERG-amplitude (R2 = 0.82, slope = 0.72, P < 0.0001). There is no significant difference between the 3-minute and the prolonged mfERG recordings for conventional amplitudes and the global-flash direct response. The global flash–induced component significantly decreases with prolonged mfERG recordings.ConclusionsA 3-minute mfERG recording and a single stimulation protocol is sufficient in anesthetized pigs. Recordings should be obtained immediately after the induction of anesthesia. The effect of TTX is significant 15 minutes after injection, but is contaminated by the effect of anesthesia 90 minutes after injection. Therefore, the quality of mfERG recordings can be further improved by determining the necessary time-of-delay from intraocular injection of a drug to full effect.Translational RelevanceGeneral anesthesia is a possible source of error in mfERG recordings. Therefore, it is important to investigate the translational relevance of the results to mfERG recordings in children in general anesthesia.

Population Response Propagation to Extrastriate Areas Evoked by Intracortical Electrical Stimulation in V1.

The mouse visual system has multiple extrastriate areas surrounding V1 each with a distinct representation of the visual field and unique functional and connectivity profiles, which are believed to form two parallel processing streams, similar to the ventral and dorsal streams in primates. At the same time, mouse visual areas have a high degree of interconnectivity, in particular V1 sends input to all higher visual areas. The study of these direct connections can further our understanding of the cortical processing of visual signals in the early mammalian cortex. Several studies have been published about the anatomy of these connections, but an in vivo electrophysiological characterization and comparison of the transmission to multiple extrastriate areas has not yet been reported. We used intracortical electrical stimulation combined with RH1691 VSD imaging in adult C57BL/6 mice in urethane anesthesia to analyze interareal transmission from V1 to extrastriate areas in superficial cortical layers. We found seven extrastriate response sites (five lateral, two medial) in a spatial pattern similar to area maps of the mouse visual cortex and, by shifting the location of V1 stimulation, demonstrated that the evoked responses in LM and AL were in accordance with the visuotopic mappings of these areas known from anatomy and in vivo studies. These two sites, considered to be gateways to their processing streams, had shorter latencies and faster transmission speeds than other extrastriate response sites. Short latency differences between response sites, and that TTX injection into LM reduced but did not eliminate other extrastriate responses indicated that the evoked cortical activity was, at least partially, transmitted directly from V1 to extrastriate areas. This study reports on analysis of interareal transmission from V1 to multiple extrastriate areas in mouse using intracortical electrical stimulation in vivo.

Myelinated Afferents Are Involved in Pathology of the Spontaneous Electrical Activity and Mechanical Hyperalgesia of Myofascial Trigger Spots in Rats.

Myofascial trigger points (MTrPs) are common causes for chronic pain. Myelinated afferents were considered to be related with muscular pain, and our clinical researches indicated they might participate in the pathology of MTrPs. Here, we applied myofascial trigger spots (MTrSs, equal to MTrPs in human) of rats to further investigate role of myelinated afferents. Modified pyridine-silver staining revealed more nerve endings at MTrSs than non-MTrSs (P < 0.01), and immunohistochemistry with Neurofilament 200 indicated more myelinated afferents existed in MTrSs (P < 0.01). Spontaneous electrical activity (SEA) recordings at MTrSs showed that specific block of myelinated afferents in sciatic nerve with tetrodotoxin (TTX) led to significantly decreased SEA (P < 0.05). Behavioral assessment showed that mechanical pain thresholds (MPTs) of MTrSs were lower than those of non-MTrSs (P < 0.01). Block of myelinated afferents by intramuscular TTX injection increased MPTs of MTrSs significantly (P < 0.01), while MPTs of non-MTrSs first decreased (P < 0.05) and then increased (P > 0.05). 30 min after the injection, MPTs at MTrSs were significantly lower than those of non-MTrSs (P < 0.01). Therefore, we concluded that proliferated myelinated afferents existed at MTrSs, which were closely related to pathology of SEA and mechanical hyperalgesia of MTrSs.

Loss of auditory activity modifies the location of potassium channel KCNQ5 in auditory brainstem neurons.

KCNQ5/Kv7.5, a low-threshold noninactivating voltage-gated potassium channel, is preferentially targeted to excitatory endings of auditory neurons in the adult rat brainstem. Endbulds of Held from auditory nerve axons on the bushy cells of the ventral cochlear nucleus (VCN) and calyces of Held around the principal neurons in the medial nucleus of the trapezoid body (MNTB) are rich in KCNQ5 immunoreactivity. We have previously shown that this synaptic distribution occurs at about the time of hearing onset. The current study tests whether this localization in excitatory endings depends on the peripheral activity carried by the auditory nerve. Auditory nerve activity was abolished by cochlear removal or intracochlear injection of tetrodotoxin (TTX). Presence of KCNQ5 was analyzed by immunocytochemistry, Western blotting, and quantitative reverse transcription polymerase chain reaction. After cochlear removal, KCNQ5 immunoreactivity was virtually undetectable at its usual location in endbulbs and calyces of Held in the anteroventral CN and in the MNTB, respectively, although it was found in cell bodies in the VCN. The results were comparable after intracochlear TTX injection, which drastically reduced KCNQ5 immunostaining in MNTB calyces and increased immunolabeling in VCN cell bodies. Endbulbs of Held in the VCN also showed diminished KCNQ5 labeling after intracochlear TTX injection. These results show that peripheral activity from auditory nerve afferents is necessary to maintain the subcellular distribution of KCNQ5 in synaptic endings of the auditory brainstem. This may contribute to adaptations in the excitability and neurotransmitter release properties of these presynaptic endings under altered input conditions. © 2014 The Authors. Journal of Neuroscience Research Published by Wiley Periodicals, Inc.

Heterogeneous firing behavior during ictal-like epileptiform activity in vitro

Seizure activity in vivo is caused by populations of neurons displaying a high degree of variability in activity pattern during the attack. The reason for this variability is not well understood. Here we show in an in vitro preparation that hippocampal CA1 pyramidal cells display four types of afterdischarge behavior during stimulus-induced ictal-like events in the presence of Cs(+) (5 mM): type I (43.7%) consisting of high-frequency firing riding on a plateau potential; type II (28.2%) consisting of low-frequency firing with no plateau potential; type III (18.3%) consisting of high-frequency firing with each action potential preceded by a transient hyperpolarization and time-locked to population activity, no plateau potential; "passive" (9.9%) typified by no afterdischarge. Type I behavior was blocked by TTX (0.2 μM) and intracellular injection of QX314 (12.5-25 mM). TTX (0.2 μM) or phenytoin (50 μM) terminated ictal-like events, suggesting that the persistent Na(+) current (I(NaP)) is pivotal for type I behavior. Type I behavior was not correlated to intrinsic bursting capability. Blockade of the M current (I(M)) with linopirdine (10 μM) increased the ratio of type I neurons to 100%, whereas enhancing I(M) with retigabine (50-100 μM) greatly reduced the epileptiform activity. These results suggest an important role of I(M) in determining afterdischarge behavior through control of I(NaP) expression. We propose that type I neurons act as pacemakers, which, through synchronization, leads to recruitment of type III neurons. Together, they provide the "critical mass" necessary for ictogenesis to become regenerative.

Comparing retinal reflectance changes elicited by transcorneal electrical retinal stimulation with those of optic chiasma stimulation in cats

To compare retinal reflectance changes (RCs) elicited by transcorneal electrical stimulation (TES) to those elicited by electrical stimulation of the optic chiasma (OX). Two eyes of two cats were studied under general anesthesia. Biphasic electrical pulses at 20 Hz were applied for 5 ms in TES, and monophasic pulses were applied for 50 μs at 100 Hz in OX stimulation. Fundus images observed with near-infrared light (800-880 nm) were recorded every 25 ms for 26 s, beginning 2 s before and ending 20 s after the electrical stimulation. To improve the signal-to-noise ratio, the images of ten consecutive recordings were averaged. Two-dimensional topographic maps of the RCs were constructed by subtracting images before the stimulation from those after the stimulation. The electrically evoked potentials (EEPs) were recorded at the OX. The effect of an intravitreal injection of tetrodotoxin (TTX) on the RCs elicited by electrical stimulation was also determined. After electrical stimulation, RCs were observed at the optic disc, retinal arteries, and retinal veins. The two-dimensional maps of the RCs elicited by both TES and OX stimulation were similar. The latency of the RCs ranged from 2.0 to 4.0 s, and the peak occurred 6 to 9 s after the onset of the ES. The intensity of the RCs was correlated with the amplitude of EEP elicited by TES stimulation. The RCs disappeared after the TTX injection in both TES and OX stimulation. TES activates principally the retinal ganglion cells, and a change in the blood flow is initiated thereafter.

Synaptic Scaling and the Development of a Motor Network

Neurons respond homeostatically to chronic changes in network activity with compensatory changes such as a uniform alteration in the size of miniature postsynaptic current (mPSC) amplitudes termed synaptic scaling. However, little is known about the impact of synaptic scaling on the function of neural networks in vivo. We used the embryonic zebrafish to address the effect of synaptic scaling on the neural network underlying locomotion. Activity was decreased during development by TTX injection to block action potentials or CNQX injection to block glutamatergic transmission. Alternatively TNFalpha was chronically applied. Recordings from spinal neurons showed that glutamatergic mPSCs scaled up approximately 25% after activity reduction and fortuitously scaled down approximately 20% after TNFalpha treatment, and were unchanged following blockade of neuromuscular activity alone with alpha-bungarotoxin. Regardless of the direction of scaling, immediately following reversal of treatment no chronic effect was distinguishable in motoneuron activity patterns or in swimming behavior. We also acutely induced a similar increase of glutamatergic mPSC amplitudes using cyclothiazide to reduce AMPA receptor desensitization or decrease of glutamatergic mPSC amplitudes using a low concentration of CNQX to partially block AMPA receptors. Though the strength of the motor output was altered, neither chronic nor acute treatments disrupted the patterning of synaptic activity or swimming. Our results show, for the first time, that scaling of glutamatergic synapses can be induced in vivo in the zebrafish and that synaptic patterning is less plastic than synaptic strength during development.

Different vulnerability in female's spatial behaviour after unilateral hippocampal inactivation

This study was aimed to determine the effect of unilateral hippocampal inactivation on spatial memory retrieval in male and female Wistar rats. Subjects were chronically implanted with a stainless-steel cannula that enabled unilateral intrahippocampal injections of tetrodotoxin, a sodium channel blocker. Rats were trained during eight consecutive days on the Morris water maze task. On day 1, subjects received a visible platform session. From day 2 to day 7 the hidden platform was located in North quadrant and on day 8 the hidden platform changed to the opposite location. Subjects were randomly distributed into four groups: male saline, male TTX, female saline, female TTX. Saline or TTX was applied 30min before training but only on days 7 and 8. Results showed that males outperform females during the initial training (days 2-6). After TTX injection, both male and female rats were impaired. However, a detailed analysis revealed that the impairment in females was more pronounced, both during the retrieval session (day 7) as well as during reversal (day 8). This data points to a different role of the hippocampus or a different distribution of memories in both sexes.

Difference in sensory dependence of occ1/ Follistatin-related protein expression between macaques and mice

occ1/ Follistatin-related protein ( Frp) is strongly expressed in the primary visual cortex (V1) of macaque monkeys, and its expression is strongly down-regulated by intraocular tetrodotoxin (TTX) injection. The pronounced area selectivity of occ1/ Frp mRNA expression occurs in macaques and marmosets, but not in mice, rabbits and ferrets, suggesting that occ1/ Frp is an important clue to the evolution of the primate cerebral cortex. To further determine species differences, we examined the sensory-input dependency of occ1/ Frp mRNA expression in mice in comparison with macaque V1. In macaque V1, occ1/ Frp mRNA expression level significantly decreased with even 1-day monocular deprivation (MD) by TTX injection. In contrast to that in macaques, however, the occ1/ Frp mRNA expression in the visual cortex in mice was not down-regulated by 1- to 7-day MD by TTX injection. Similarly, MD had no effect on occ1/ Frp mRNA expression level in the dorsal lateral geniculate nucleus of mice. In addition, the extirpation of the cochlear or olfactory epithelium had no effect on occ1/ Frp mRNA expression in either the cochlear nucleus or the olfactory bulb in mice. Thus, occ1/ Frp mRNA expression is independent of sensory-input in mice. The results suggest that activity-dependent occ1/ Frp mRNA expression is not common between mice and monkeys, and that primate V1 has acquired a unique gene regulatory mechanism that enables a rapid response to environmental changes. The characteristic feature of the activity dependency of occ1/ Frp mRNA expression is discussed, in comparison with that of the expression of the immediate-early genes, c-fos and zif268.

Episodic vestibular disruption following ablation of the inferior olive in rats: Behavioral correlates

The experiments herein investigate whether the behavioral responses to transient and episodic vestibular disruption and permanent ablation are distinct in the absence of climbing fiber input. Subjects in group 1 received an IP injection of PBS followed by an IP injection of niacinamide. Seven days later these rats received the first of 3 serial transtympanic injections of TTX on the same side with 7 days between each injection. Following each TTX injection rats displayed unilateral vestibular symptoms that persisted beyond 48 h. Spontaneous barrel rolling behavior was not observed. Group 2 subjects received an IP injection of 3-acetylpyridine (3-AP) + niacinamide followed by the same TTX regimen as group 1. Following each TTX injection vestibular symptoms (severe body twisting and persistent spontaneous barrel rolling) emerged rapidly (<15 min) and resolved by 72 h. Group 3 subjects received an IP injection of 3-AP + niacinamide and 7 days later a single unilateral transtympanic injection of sodium arsanilate. Rats in group 3 developed vestibular symptoms similar to those observed in group 2 although there was no resolution of these symptoms. The results indicate that TTX has a rapid rate of infiltration and blockade of the VIIIth nerve that persists for >48 h and then completely resolves. The contrast in vestibular symptoms between groups 1 and 2 suggest that climbing fibers are recruited soon after onset of vestibular disruption and play a role in attenuating the severity of vestibular symptoms associated with transient/episodic vestibular disruption.

Antinociceptive effects of tetrodotoxin (TTX) in rodents

Tetrodotoxin (TTX) is a powerful sodium channel blocker extracted from the puffer fish. The analgesic effects of TTX were investigated in different animal pain models. Wistar rats were submitted to the formalin test and to partial ligation of the sciatic nerve (Seltzer's model). Swiss Webster mice were used in the writhing test. Rodents were divided into six groups receiving a s.c. injection of either 0.9% NaCl, TTX 0.3, 1, 3, or 6 microg kg(-1), or morphine (5 mg kg(-1)). Substances were injected 30 min before 2.5% formalin injection into the hind paw, acetic acid administration intraperitoneally or neuropathic pain testing consisting of mechanical allodynia (von Frey filament) and thermal hyperalgesia (Plantar test). TTX decreased pain behaviour in the formalin test at the highest dose and in the writhing test at 3 and 6 microg kg(-1). It also diminished mechanical allodynia and thermal hyperalgesia with an ED(50) of 1.08 (0.89) and 0.62 (0.33) microg kg(-1), respectively. Observation of the rats after TTX injection did not show any motor deficit, respiratory distress or sedation. Morphine was also effective in relieving pain in all three tests but with signs of considerable sedation. Systemic injections of TTX diminished pain behaviour in a dose-dependent manner in models of inflammatory, visceral and neuropathic pain without causing adverse events, whereas morphine analgesia was associated with heavy sedation. TTX is a very promising substance for the treatment of various types of pain but needs further evaluation.

Slow oscillatory activity of rat globus pallidus neurons in vitro

The neurons in the external segment of the pallidum in the primate develop a characteristic firing pattern consisting of alternately occurring long, 2-20 s, strongly active phases and long completely silent phases when the subthalamo-pallidal excitatory inputs are blocked. The induction of the activity might be a factor in the development of dyskinesias after the loss of subthalamic output. In this study, we used globus pallidus (GPe) slice preparations obtained from juvenile rats to examined the conditions that support the alternatively occurring long depolarized and hyperpolarized phases which we refer to as the slow oscillation (SO). SO was not induced by the blockade of glutamatergic inputs but was induced by treatments that depolarized dendrites and, at the same time, hyperpolarized the somata with current injections. The treatments included elevation of extracellular K(+), application of K-current blockers and the lowering of extracellular Ca(2+). Application of TTX or intracellular BAPTA injection blocked the SO, while the SO could be maintained in hyperpolarization-activated inward current blockers, organic Ca-current blockers and up to 200 microm CdCl(2). These results suggest that Na currents play a major role in the generation of SO in vitro. It can be speculated that Na currents are involved in the development of active phases observed in the GPe after blockade of the glutamatergic inputs in vivo and that the unique property of GPe neurons in maintaining strong activity after the elimination of the glutamatergic driving force contributes to the development of motor disorders such as dyskinesia.

Behavioral Evidence That Segregation and Representation Are Dissociable Hippocampal Functions

Hippocampal activity is thought to encode spatial representations in a distributed associative network. This idea predicts that partial hippocampal lesions would spare acquisition and impair retrieval of a place response as long as enough connections remained intact to encode associations. Water maze experiments supported the predictions, but the prediction of impaired retrieval was not supported when tetrodotoxin (TTX) was injected into one hippocampus and rats were tested in a place avoidance task on a rotating arena with shallow water. The rotation dissociated relevant distal stimuli from irrelevant self-motion stimuli. To explain the discrepancy, we hypothesized that the segregation of relevant and irrelevant stimuli and stimuli association into representations are distinct hippocampus-dependent operations, and whereas associative representation is more sensitive to disruption during retrieval than learning, stimulus segregation is more sensitive to disruption during learning than during retrieval. The following predictions were tested: (1) the TTX injection would spare learning but (2) impair retrieval of a place response in the water maze, which has a high associative representational demand but a low demand for segregation; (3) the injection would impair learning but (4) spare retrieval of place avoidance in the rotating arena filled with water, which has a high demand for stimulus segregation but a low associative representational demand. All four predictions were confirmed. The hypothesis also explains the pattern of sparing and impairment after the TTX injection in other place avoidance task variants, leading us to conclude that stimulus separation and association representation are dissociable functions of the hippocampus.

The Decline of the Photopic Negative Response (PhNR) in the Rat after Optic Nerve Transection

To investigate the contribution to the photopic negative response (PhNR) of the electroretinogram (ERG) by retinal ganglion cells (RGCs). The PhNR was assessed longitudinally following optic nerve transection (ONTx). Photopic ERGs were recorded from each eye of an anesthetized (ketamine/xylazine, 60 mg/kg and 5 mg/kg) Brown Norway rat using custom made electrodes (PT-IR Tef., A-M System Inc). ERGs were elicited using green Ganzfeld flashes (11.38 scd/m(2), 22.76 cds/m(2)) and a rod suppressing green-background (40 cd/m(2)). PhNRs were compared before and after optic nerves were transected. Cresyl violet stained retinal flatmounts were used to estimate cell loss in the ganglion cell layer 3 and 15 weeks after optic nerve transection. The pharmacological effect of 1.3 microM intravitreal TTX on the PhNR was also evaluated. There was a significant loss (p <0.05) in the PhNR of 20, 36, 34, 35, 48, 48 and 56% for ONTx eye versus the contralateral eye, at post ONTx times of 24 h, 1, 2, 3, 4, 8 and 15 weeks. B-wave amplitudes of ONTx eyes were not significantly different from the control eyes. In ONTx eyes, mean cell loss in the retinal ganglion cell layer was 27 and 55% at the 3 week and 15 week time periods. In the eyes with ONTx, the decline of PhNR amplitudes was correlated positively with RGC loss (r = 0.98; p < 0.01). Thirty minutes after intravitreal TTX injection, the PhNR was significantly reduced (57%, p<0.01). There was a time-dependent decline in the PhNR after ONTx, as exemplified by a 35% reduction from 1-3 weeks, a 48% decline for 4-8 weeks and a 56% decline after 15 weeks. The correlation between the decline in the PhNR and retinal ganglion cell loss suggests that the PhNR depends on inner retina integrity and the PhNR may be important biological signal or detecting glaucomatous damage and the monitoring of RGC function changes in early glaucoma.

Beyond Memory, Navigation, and Inhibition: Behavioral Evidence for Hippocampus-Dependent Cognitive Coordination in the Rat

Injecting tetrodotoxin (TTX) into one hippocampus impaired avoidance of a place defined by distal cues while rats were on a slowly rotating arena. The impairment could be explained by a deficit in memory, navigation, or behavioral inhibition. Here, we show that the TTX injection abolished the ability of rats to organize place-avoidance behavior specifically when distal room and local arena cues were continuously dissociated. The results provide evidence that injecting TTX into one hippocampus specifically impaired the coordination of representations that support organized behavior because of the following: (1) rats normally coordinate separate room and arena avoidance memories; (2) the TTX injection spared spatial, relational, and representational memory, navigation, and behavioral inhibition; and (3) the TTX-induced impairment of place avoidance depended on the need to coordinate representations of local and distal stimuli.

Temporary inactivation of the nucleus accumbens disrupts acquisition and expression of fear-potentiated startle in rats

Recent research suggests that in addition to its prominent role in appetitive learning, the nucleus accumbens (NAC) may also be involved in fear conditioning. In the present study, we investigated whether temporary inactivation of the NAC, by injection of tetrodotoxin (TTX), affects acquisition and expression of conditioned fear, as measured by fear-potentiated startle (FPS). TTX injection into the NAC totally blocked acquisition and markedly decreased expression of conditioned fear to a discrete visual conditioned stimulus (CS). Interestingly, temporary inactivation of the NAC did not affect shock sensitization of startle, indicating that both the perception of the shock and short-term contextual conditioning was not affected by intra-accumbal TTX injection. Taken together, these results show that the NAC is crucial for acquisition and expression of long-term conditioned fear, as measured by fear-potentiated startle, to discrete CSs, but not short-term conditioned fear to a context.

Episodic blockade of cranial nerve VIII provokes asymmetric changes in lobule X of the rat

Although debilitating syndromes like Ménière's disease are in part characterized by recurrent or episodic vestibular disturbance the study of episodic vestibular disruption has only recently been possible with the introduction of a new model utilizing tetrodotoxin (TTX). In the present study, serial unilateral transtympanic administration of TTX produced behavioral symptoms indicative of transient vestibular disruption and novel patterns of Fos activity in the brainstem and cerebellum. Following two or three serial injections of TTX and a final survival time of 2 h, Fos immunocytochemistry revealed a distinct pattern of labeling in the brainstem that differed temporally from that observed following a single unilateral TTX injection. Specifically there was protracted expression of Fos in the β subdivision of the inferior olive (IO) on the side ipsilateral to TTX treatment. In the cerebellum, the hallmark of episodic vestibular blockade was an asymmetric pattern of Fos labeling that involved all three layers of the cortex. In particular, there was prominent Fos labeling of Purkinje cells in the contra-TTX half of lobule X. In view of the fact that Fos labeling is not found in Purkinje cells following a single transient event or following peripheral vestibular ablation, it is suggested that Fos expression in Purkinje cells is a unique feature of episodic vestibular disruption and may represent a novel plastic response by a select population of Purkinje cells to episodic functional deafferentation.

Changes in neuronal activity of the inferior colliculus in rat after temporal inactivation of the auditory cortex.

The role of cortico-tectal pathways in auditory signal processing was studied in anesthetized rats by comparing the extracellular single unit activity in the inferior colliculus (IC) before and after functional ablation of the auditory cortex (AC) by tetrodotoxin (TTX). The responses of several IC neurons to sound stimuli were simultaneously recorded with a 16-channel electrode probe introduced into the IC. Click-evoked middle latency responses (MLR) recorded from the AC were suppressed for several hours after TTX injection. During AC inactivation the firing rate of IC neurons increased (40 % of neurons), decreased (44 %) or did not change (16 %) in comparison with control conditions. In several IC neurons, TTX injection resulted in alterations in the shape of the rate-level functions. Response thresholds, tuning properties and the type of discharge pattern of IC neurons were not altered during AC inactivation. However, in one-third of the neurons, the initial part of the response was less altered than the later, sustained part. In two-thirds of neuronal pairs, functional decortication resulted in a change in the cross-correlation coefficient. The results reveal the complex changes that appear in IC neuronal activity after functional ablation of the ipsilateral auditory cortex.

Ocular dominance columns and local projections of layer 6 pyramidal neurons in macaque primary visual cortex.

To study the relationship between ocular dominance columns (ODCs) and axonal projections of individual layer 6 pyramidal neurons in the primary visual cortex, neurons were intracellularly labeled with biocytin in live slices prepared from macaque monkeys that had received an intravitreal injection of tetrodotoxin (TTX). The TTX injection indirectly causes a decrease in cytochrome oxidase (CO) expression in the cortical ODCs corresponding to the treated eye (Wong-Riley & Carroll, 1984). Sections from slices with labeled layer 6 neurons were double stained for biocytin and CO, to allow visualization of neuronal processes as well as ODCs. Twenty-seven layer 6 pyramidal neurons in ODC-labeled slices were analyzed. These neurons were classified according to the criteria of Wiser and Callaway (1996). Eight of these are class I neurons, which have dense axonal projections to the monocular layer 4C. The remaining 19 are class II neurons which project primarily to the binocular layers outside 4C. Among class I neurons, two have dense axonal arbors in layer 4C alpha (type I alpha), one in layer 4C beta (type I beta), and two throughout the depth of layer 4C (type IC). None of these neurons have ODC-specific axonal arbors. The remaining three class I neurons have focused axonal projections in layers 4C beta and 4A (type I beta A). All three appear to have axonal arbors predominantly within their home ODC in layer 4C. The axonal arbors of class II neurons do not appear to relate to ODCs in any specific fashion.