Long-term cortical plasticity evoked by electric stimulation and acetylcholine applied to the auditory cortex

Auditory fear conditioning with tone bursts followed by electric leg stimulation activates neurons not only in the auditory and somatosensory systems but also in many other regions of the brain and elicits shifts in the best frequencies (BFs) of collicular and cortical neurons, i.e., reorganization of the frequency (co-chleotopic) maps in the inferior colliculus and auditory cortex (AC). What are the neural elements minimally necessary for evoking long-term cortical BF shifts? We found that: (i) both electric stimulation and acetylcholine applied to the AC evoke the long-term cortical BF shift as does the conditioning; (ii) both electric stimulation of the AC and acetylcholine applied to the inferior colliculus increase the short-term collicular BF shift evoked by the cortical electric stimulation but do not change it into long-term; and (iii) as this short-term collicular BF shift is blocked by atropine, the development of the long-term cortical BF shift becomes slow and small. Therefore, the most essential neural elements for evoking the long-term cortical BF shift are the AC, corticofugal feedback and the cholinergic nucleus. Our current data support the Gao-Suga model, which hypothesizes that the small short-term cortical BF shifts are evoked by tonal stimuli without the association of conditioned and unconditioned stimuli in the multisensory thalamic nuclei and that these BF shifts are augmented and changed into the large long-term BF shifts by cholinergic neurons.     

Asymmetry in corticofugal modulation of frequency-tuning in mustached bat auditory system

Focal electric stimulation of the auditory cortex is well suited for exploration of the function of the corticofugal (descending) system and the neural mechanism of plasticity in the central auditory system, because it evokes changes in frequency-tuning, called best frequency (BF) shifts, as does auditory fear conditioning. The Doppler-shifted constant frequency (DSCF) area of the primary auditory cortex of the mustached bat is highly specialized for fine frequency analysis. Focal electric stimulation of the DSCF area evokes the BF shifts of ipsilateral cortical and collicular neurons away from the BF of stimulated neurons, whereas the stimulation evokes the BF shifts of contralateral cortical and collicular neurons either toward or away from the stimulated BF. The direction of contralateral BF shifts shows a flip-flop, depending on the spatial relationship between the stimulated and recorded neurons. This asymmetry in corticofugal modulation is mostly, if not totally, created by two subdivisions of the stimulated DSCF area that transmit signals to the contralateral DSCF area, presumably through the corpus callosum. This intriguing asymmetry in corticofugal modulation presumably functions for equalization of the reorganization of the frequency maps of the DSCF areas and subcortical auditory nuclei on both sides.     

Plasticity of the cochleotopic (frequency) map in specialized and nonspecialized auditory cortices

Auditory conditioning (associative learning) causes reorganization of the cochleotopic (frequency) maps of the primary auditory cortex (AI) and the inferior colliculus. Focal electric stimulation of the AI also evokes basically the same cortical and collicular reorganization as that caused by conditioning. Therefore, part of the neural mechanism for the plasticity of the central auditory system caused by conditioning can be explored by focal electric stimulation of the AI. The reorganization is due to shifts in best frequencies (BFs) together with shifts in frequency-tuning curves of single neurons. In the AI of the Mongolian gerbil (Meriones unguiculatus) and the posterior division of the AI of the mustached bat (Pteronotus parnellii), focal electric stimulation evokes BF shifts of cortical auditory neurons located within a 0.7-mm distance along the frequency axis. The amount and direction of BF shift differ depending on the relationship in BF between stimulated and recorded neurons, and between the gerbil and mustached bat. Comparison in BF shift between different mammalian species and between different cortical areas of a single species indicates that BF shift toward the BF of electrically stimulated cortical neurons (centripetal BF shift) is common in the AI, whereas BF shift away from the BF of electrically stimulated cortical neurons (centrifugal BF shift) is special. Therefore, we propose a hypothesis that reorganization, and accordingly organization, of cortical auditory areas caused by associative learning can be quite different between specialized and nonspecialized (ordinary) areas of the auditory cortex.     

Reorganization of the cochleotopic map in the bat's auditory system by inhibition

The central auditory system of the mustached bat shows two types of reorganization of cochleotopic (frequency) maps: expanded reorganization resulting from shifts in the best frequencies (BFs) of neurons toward the BF of repetitively stimulated cortical neurons (hereafter centripetal BF shifts) and compressed reorganization resulting from the BF shifts of neurons away from the BF of the stimulated cortical neurons (hereafter centrifugal BF shifts). Facilitation and inhibition evoked by the corticofugal system have been hypothesized to be respectively related to centripetal and centrifugal BF shifts. If this hypothesis is correct, bicuculline (an antagonist of inhibitory GABA-A receptors) applied to cortical neurons would change centrifugal BF shifts into centripetal BF shifts. In the mustached bat, electric stimulation of cortical Doppler-shifted constant-frequency neurons, which are highly specialized for frequency analysis, evokes the centrifugal BF shifts of ipsilateral collicular and cortical Doppler-shifted constant-frequency neurons and contralateral cochlear hair cells. Bicuculline applied to the stimulation site changed the centrifugal BF shifts into centripetal BF shifts. On the other hand, electric stimulation of neurons in the posterior division of the auditory cortex, which are not particularly specialized for frequency analysis, evokes centripetal BF shifts of cortical neurons located near the stimulated cortical neurons. Bicuculline applied to the stimulation site augmented centripetal BF shifts but did not change the direction of the shifts. These observations support the hypothesis and indicate that centripetal and centrifugal BF shifts are both based on a single mechanism consisting of two components: facilitation and inhibition.     

Corticofugal modulation of duration-tuned neurons in the midbrain auditory nucleus in bats

Animal sounds, as well as human speech sounds, are characterized by multiple parameters such as frequency, intensity, duration, etc. The central auditory system produces neurons tuned to particular durations and frequencies of sounds emitted by a species. In bats, "duration-tuned" neurons are mostly sensitive to short durations and high frequencies of sounds used for echolocation. They are scattered in the frequency maps of the inferior colliculus and auditory cortex. We found that electric stimulation of cortical duration-tuned neurons modulates collicular duration-tuned neurons in both duration and frequency tuning only when collicular and cortical neurons paired for studies are within +/-4 ms in best duration and within +/-6 kHz in best frequency. There are four types of modulations: sharpening or broadening of duration tuning, and lengthening or shortening of best duration. Sharpening is observed in "matched" collicular neurons whose best durations are the same as those of stimulated cortical neurons, and it is accompanied by augmentation of the auditory responses at their best durations. The other three types of modulations are observed in "unmatched" collicular neurons whose best durations are different from those of stimulated cortical neurons. Lengthening or shortening of best duration is linearly related to the amount of the difference in best duration between collicular and cortical neurons. Corticofugal modulation is specific and systematic according to relationships in both duration and frequency between stimulated cortical and recorded collicular neurons.     

Centripetal and centrifugal reorganizations of frequency map of auditory cortex in gerbils

As repetitive acoustic stimulation and auditory conditioning do, electric stimulation of the primary auditory cortex (AI) evokes reorganization of the frequency map of AI, as well as of the subcortical auditory nuclei. The reorganization is caused by shifts in best frequencies (BFs) of neurons either toward (centripetal) or away from (centrifugal) the BF of stimulated cortical neurons. In AI of the Mongolian gerbil, we found that focal electrical stimulation evoked a centripetal BF shift in an elliptical area centered at the stimulated neurons and a centrifugal BF shift in a zone surrounding it. The 1.9-mm long major and 1.1-mm long minor axes of the elliptical area were parallel and orthogonal to the frequency axis, respectively. The width of the surrounding zone was 0.2-0.3 mm. Such "center-surround" reorganization has not yet been found in any sensory cortex except AI of the gerbil. The ellipse is similar to the arborization pattern of pyramidal neurons, the major source of excitatory horizontal connections in AI, whereas the surrounding zone is compatible to the arborization range of small basket cells (inhibitory neurons) in AI.     

The corticofugal system for hearing: recent progress

Peripheral auditory neurons are tuned to single frequencies of sound. In the central auditory system, excitatory (or facilitatory) and inhibitory neural interactions take place at multiple levels and produce neurons with sharp level-tolerant frequency-tuning curves, neurons tuned to parameters other than frequency, cochleotopic (frequency) maps, which are different from the peripheral cochleotopic map, and computational maps. The mechanisms to create the response properties of these neurons have been considered to be solely caused by divergent and convergent projections of neurons in the ascending auditory system. The recent research on the corticofugal (descending) auditory system, however, indicates that the corticofugal system adjusts and improves auditory signal processing by modulating neural responses and maps. The corticofugal function consists of at least the following subfunctions. (i) Egocentric selection for short-term modulation of auditory signal processing according to auditory experience. Egocentric selection, based on focused positive feedback associated with widespread lateral inhibition, is mediated by the cortical neural net working together with the corticofugal system. (ii) Reorganization for long-term modulation of the processing of behaviorally relevant auditory signals. Reorganization is based on egocentric selection working together with nonauditory systems. (iii) Gain control based on overall excitatory, facilitatory, or inhibitory corticofugal modulation. Egocentric selection can be viewed as selective gain control. (iv) Shaping (or even creation) of response properties of neurons. Filter properties of neurons in the frequency, amplitude, time, and spatial domains can be sharpened by the corticofugal system. Sharpening of tuning is one of the functions of egocentric selection.     

Laterality effects of human pudendal nerve stimulation on corticoanal pathways: evidence for functional asymmetry

BACKGROUND: Although motor and sensory pathways to the human external anal sphincter are bilateral, a unilateral pudendal neuropathy may still disrupt anal continence. Anal continence can, however, be preserved despite unilateral pudendal damage, and so to explain those differing observations, we postulated that pudendal innervation might be asymmetric. AIMS: To explore the individual effects of right and left pudendal nerve stimulation on the corticofugal pathways to the human external anal sphincter and thus assess evidence for functional asymmetric pelvic innervation. METHODS: In eight healthy subjects, anal sphincter electromyographic responses, evoked to transcranial magnetic stimulation of the motor cortex, were recorded 5-500 msec after digital transrectal electrical conditioning stimuli applied to each pudendal nerve. RESULTS: Right or left pudendal nerve stimulation evoked anal responses of similar latencies but asymmetric amplitudes in six subjects: dominant responses (>50% contralateral side) from the right pudendal in four subjects and from the left in two. Cortical stimulation also evoked anal responses with amplitude 448 (121) microV and latency 20.9 (1.1) msec. When cortical stimulation was preceded by pudendal nerve stimulation, the cortical responses were facilitated at interstimulus intervals of 5-20 msec. Dominant pudendal nerve stimulation induced greater facilitation of the cortically evoked responses than the non-dominant nerve. CONCLUSIONS: Cortical pathways to the external anal sphincter are facilitated by pudendal nerve conditioning, in an asymmetric manner. This functional asymmetry may explain the presence and absence of anal incontinence after unilateral pudendal nerve injury.     

Dorsal periaqueductal gray-amygdala pathway conveys both innate and learned fear responses in rats

The periaqueductal gray (PAG) and amygdala are known to be important for defensive responses, and many contemporary fear-conditioning models present the PAG as downstream of the amygdala, directing the appropriate behavior (i.e., freezing or fleeing). However, empirical studies of this circuitry are inconsistent and warrant further examination. Hence, the present study investigated the functional relationship between the PAG and amygdala in two different settings, fear conditioning and naturalistic foraging, in rats. In fear conditioning, electrical stimulation of the dorsal PAG (dPAG) produced unconditional responses (URs) composed of brief activity bursts followed by freezing and 22-kHz ultrasonic vocalization. In contrast, stimulation of ventral PAG and the basolateral amygdalar complex (BLA) evoked freezing and/or ultrasonic vocalization. Whereas dPAG stimulation served as an effective unconditional stimulus for fear conditioning to tone and context conditional stimuli, neither ventral PAG nor BLA stimulation supported fear conditioning. The conditioning effect of dPAG, however, was abolished by inactivation of the BLA. In a foraging task, dPAG and BLA stimulation evoked only fleeing toward the nest. Amygdalar lesion/inactivation blocked the UR of dPAG stimulation, but dPAG lesions did not block the UR of BLA stimulation. Furthermore, in vivo recordings demonstrated that electrical priming of the dPAG can modulate plasticity of subiculum–BLA synapses, providing additional evidence that the amygdala is downstream of the dPAG. These results suggest that the dPAG conveys unconditional stimulus information to the BLA, which directs both innate and learned fear responses, and that brain stimulation-evoked behaviors are modulated by context.     

Changes in parvalbumin immunoreactivity with aging in the central auditory system of the rat

Changes in the levels of calcium binding proteins are known to occur in different parts of the brain during aging. In our study we attempted to define the effect that aging has on the parvalbumin-expressing system of neurons in the higher parts of the central auditory system. Age-related changes in parvalbumin immunoreactivity were investigated in the inferior colliculus (IC), medial geniculate body (MGB) and auditory cortex (AC) in two rat strains, normally aging Long-Evans (LE) and fast aging Fischer 344 (F344). The results demonstrate that the changes in PV-immunoreactivity are strain-dependent with an increase in the number of PV-immunoreactive (PV-ir) neurons occurring in the inferior colliculus of old LE rats and a pronounced decline in the number of PV-ir neurons appearing in the auditory cortex of aged F344 animals. In some parts of the AC of old F344 animals no PV-ir neurons were present at all. The number of PV-ir neurons in the MGB in all examined animals was very low independent of the strain and age. The loss of PV-ir neurons in the auditory cortex of Fischer 344 rats with aging may contribute to the substantial deterioration of hearing function in this strain.     

Corticothalamic synchronization leads to c-fos expression in the auditory thalamus

In this study, we investigated the relationship between c-fos expression in the auditory thalamus and corticofugal activation. The contribution of neurotransmitters and related receptors, the involvement of thalamic reticular nucleus (TRN), and the role of neuronal firing patterns in this process were also examined. The principal nuclei of the medial geniculate body (MGB) showed c-fos expression when the auditory cortex (AC) was activated by direct injection of bicuculline methobromide. However, no expression was detectable with acoustic stimuli alone. This indicated that c-fos expression in the principal nuclei of the MGB was triggered by the corticofugal projection. c-fos expression could be elicited in the MGB by direct injection of glutamate. Direct administration of acetylcholine, alternatively, had no effect. Bicuculline methobromide injection in the AC also triggered synchronized oscillatory activities sequentially in the AC and MGB. Cortically induced c-fos expression in the MGB was not mediated by a pathway involving the TRN because it remained intact after a TRN lesion with kainic acid. The present results also conclude that c-fos expression is not simply associated with firing rate, but also with neuronal firing pattern. Burst firings that are synchronized with the cortical oscillations are proposed to lead to c-fos expression in the principal nuclei of the MGB.     

Long-term modifications of synaptic efficacy in the human inferior and middle temporal cortex.

The primate temporal cortex has been demonstrated to play an important role in visual memory and pattern recognition. It is of particular interest to investigate whether activity-dependent modification of synaptic efficacy, a presumptive mechanism for learning and memory, is present in this cortical region. Here we address this issue by examining the induction of synaptic plasticity in surgically resected human inferior and middle temporal cortex. The results show that synaptic strength in the human temporal cortex could undergo bidirectional modifications, depending on the pattern of conditioning stimulation. High frequency stimulation (100 or 40 Hz) in layer IV induced long-term potentiation (LTP) of both intracellular excitatory postsynaptic potentials and evoked field potentials in layers II/III. The LTP induced by 100 Hz tetanus was blocked by 50-100 microM DL-2-amino-5-phosphonovaleric acid, suggesting that N-methyl-D-aspartate receptors were responsible for its induction. Long-term depression (LTD) was elicited by prolonged low frequency stimulation (1 Hz, 15 min). It was reduced, but not completely blocked, by DL-2-amino-5-phosphonovaleric acid, implying that some other mechanisms in addition to N-methyl-DL-aspartate receptors were involved in LTD induction. LTD was input-specific, i.e., low frequency stimulation of one pathway produced LTD of synaptic transmission in that pathway only. Finally, the LTP and LTD could reverse each other, suggesting that they can act cooperatively to modify the functional state of cortical network. These results suggest that LTP and LTD are possible mechanisms for the visual memory and pattern recognition functions performed in the human temporal cortex.     

Effects of recurrent antecedent hypoglycaemia and chronic hyperglycaemia on brainstem extra-cellular glucose concentrations during acute hypoglycaemia in conscious diabetic BB rats

Aims/hypothesis Our aim was to determine whether the divergent effects of chronic exposure to hyperglycaemia or hypoglycaemia on the glycaemic threshold for auditory brainstem dysfunction are reflected in the extra-cellular fluid (ECF) concentrations of glucose in the inferior colliculus during hypoglycaemia in the diabetic BB rat.Methods Microdialysis was used to measure inferior colliculus ECF glucose concentrations under basal and hyperinsulinaemic (20 mU/kg·min) hypoglycaemic conditions.Results ECF glucose is increased under basal (hyperglycaemic) conditions and decreases during hypoglycaemia in both recurrently hypoglycaemic and chronically hyperglycaemic diabetic BB rats (to 0.5±0.1 and 0.8±0.2 mmol/L respectively), with no significant differences between groups. In both groups the plasma to ECF glucose ratio doubled during hypoglycaemia.Conclusion/interpretation Prior exposure to recurrent hypoglycaemia does not lead to increased ECF glucose concentrations in the inferior colliculus of diabetic BB rats. The resistance to impaired brainstem function seen in recurrently hypoglycaemic rats during hypoglycaemia cannot simply be attributed to increased blood-brain barrier glucose transport within this brain region.Abbreviations BBB Blood-brain barrier - RH recurrent hypoglycaemia - BAEP brainstem auditory evoked potentials - IC inferior colliculus - CH chronic hyperglycaemia - ECF extra-cellular fluid     

Seizures during Ethanol Withdrawal Are Blocked by Focal Microinjection of Excitant Amino Acid Antagonists into the Inferior Colliculus and Pontine Reticular Formation

Physical dependence on ethanol can result in seizure susceptibility during ethanol withdrawal. In rats, generalized tonic-clonic seizures are precipitated by auditory stimulation during the ethanol withdrawal syndrome. Excitant amino acids (EAAs) are implicated as neurotransmitters in the inferior colliculus and the brain stem reticular formation, which play important roles in the neuronal network for genetic models of audiogenic seizures (AGSs). Ethanol blocks the actions of EAAs in various brain regions, including the inferior colliculus. In this study, dependence was produced by intragastric administration of ethanol for 4 days. During ethanol withdrawal, AGSs were blocked by systemic administration of competitive or noncompetitive NMDA antagonists 3-((±)-2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP) or dizocilpine (MK-801). Focal microinjections of NMDA or non-NMDA antagonists into the inferior colliculus or the pontine reticular formation also inhibited AGSs. MK-801 was the most potent anticonvulsant systemically. When injected into the inferior colliculus, CPP had a more potent anticonvulsant effect than either MK-801 or the non-NMDA antagonist 6-cyano-7-nitroquinoxa-line-2,3-dione. The inferior colliculus was more sensitive than the pontine reticular formation to the anticonvulsant effects of both competitive NMDA and non-NMDA antagonists. The results of the present support the idea that continued ethanol administration may lead to development of supersensitivity to the action of EAAs in inferior colliculus and pontine reticular formation neurons. This may be a critical mechanism subserving AGS susceptibility during ethanol withdrawal.     

Long-term retention of learning-induced receptive-field plasticity in the auditory cortex.

Brief learning experience (classical conditioning) induces frequency-specific receptive-field (RF) plasticity in the auditory cortex, characterized as increased response to the frequency of the conditioned stimulus and decreased responses to most other frequencies, including the pretraining best frequency. This experiment asked (i) whether learning-induced RF plasticity, established in the waking state, can be expressed under general anesthesia and if so (ii) whether it exhibits long-term retention. Pure-tone-frequency RFs were obtained from adult guinea pigs under general anesthesia (sodium pentobarbital or ketamine) before and repeatedly after (1 hr-8 weeks) a 20- to 30-trial session of pairing a non-best-frequency tone with mild footshock. Conditioned-stimulus-specific RF plasticity was expressed under both types of anesthesia and included shifts of the pretraining best frequency toward or even to the frequency of the conditioned stimulus. Moreover, this RF plasticity exhibits long-term retention, being evident 1-8 weeks after training. This satisfies a criterion for the long-term storage of information in the auditory cortex.     

Modulation of auditory processing by cortico-cortical feed-forward and feedback projections

The auditory center in the cerebrum, the auditory cortex, consists of multiple interconnected areas. The functional role of these interconnections is poorly understood. The auditory cortex of the mustached bat consists of at least nine areas, including the frequency modulation-frequency modulation (FF) and dorsal fringe (DF) areas. The FF and DF areas consist of neurons tuned to specific echo delays carrying target-distance information. The DF area is hierarchically at a higher level than the FF area. Here, we show that the feedback projection from the DF area to the FF area shifts the delay-tuning of FF neurons toward that of the stimulated DF neurons. In contrast, the feed-forward projection from the FF area to the DF area shifts the delay-tuning of DF neurons away from that of the stimulated FF neurons. The lateral projection within the DF area shifts the delay-tuning of DF neurons toward that of the stimulated DF neurons. In contrast, the lateral projection within the FF area shifts the delay-tuning of FF neurons away from that of the stimulated FF neurons. The delay-tuning shift evoked by the DF stimulation was 2.5 times larger than that evoked by the FF stimulation. Our data indicate that the FF-DF feed-forward and FF-FF lateral projections shape the highly selective neural representation of the tuning of the excited DF neurons, whereas the DF-FF feedback and DF-DF lateral projections enhance the representation of the selected tuning, perhaps, for focal processing of information carried by the excited FF neurons.     

Multiple Withdrawals from Chronic Ethanol "Kindles" Inferior Collicular Seizure Activity: Evidence for Kindling of Seizures Associated with Alcoholism

The present investigation tested the hypothesis that multiple withdrawals from chronic ethanol treatment "kindles" seizure activity. Two animal models of kindled seizure activity--electrical stimulation of the inferior collicular cortex or the amygdala--were used to evaluate this hypothesis. Four withdrawals from a 12-day ethanol-liquid diet regimen facilitated the seizure kindling rate in the inferior collicular cortex, when the stimulation was initiated 7 days after the last withdrawal. In contrast, four withdrawals from this chronic ethanol regimen significantly attenuated the rate of amygdaloid kindling. When the withdrawals were increased to six or 10 using a 5-day chronic ethanol treatment schedule, the kindling rate in the inferior collicular cortex proved directly proportional to the withdrawal number. Continuous ethanol exposure over the same period as the 10 withdrawal group also facilitated the inferior collicular kindling rate, but not to the extent found in the 10 withdrawal group. A before, 10 withdrawals from the 5-day chronic ethanol liquid diet treatment attenuated the rate of amygdaloid kindling. Thus, this kindling action of repeated ethanol withdrawals appears specific to seizures originating from the inferior collicular cortex, not the limbic system. These findings support a previous hypothesis for a kindling etiology of alcoholism related seizures.     

Visual- and saccade-related signals in the primate inferior colliculus

The inferior colliculus (IC) is normally thought of as a predominantly auditory structure because of its early position in the ascending auditory pathway just before the auditory thalamus. Here, we show that a majority of IC neurons (64% of 180 neurons) in awake monkeys carry visual- and/or saccade-related signals in addition to their auditory responses (P < 0.05). The response patterns involve primarily excitatory visual responses, but also increased activity time-locked to the saccade, slow rises in activity time-locked to the onset of the visual stimulus, and inhibitory responses. The presence of these visual-related signals suggests that the IC plays a role in integrating visual and auditory information. More broadly, our results show that interactions between sensory pathways can occur at very early points in sensory processing streams, which implies that multisensory integration may be a low-level rather than an exclusively high-level process.     

A Potential Contribution to Ethanol Withdrawal Kindling: Reduced GABA Function in the Inferior Collicular Cortex

Because multiple withdrawals from chronic ethanol treatment facilitate the rate of kindling from the inferior collicular cortex, the following studies sought to identity potential sources of this long-term change in seizure sensitivity. When rats received 6 or 10 withdrawals from a 5-day ethanol liquid diet, a significant decrease was found in the threshold frequency for seizure genesis, 6–7 days postwithdrawal. The magnitude of this change was related to the number of withdrawals (6 withdrawals, -2.0 ± 0.3 Hz; 10 withdrawals, -3.2 ± 1.2 Hz). Thus, multiple ethanol withdrawals increased seizure sensitivity within the inferior collicular cortex. On the following day in the same animals, changes in inhibitory or excitatory function were evaluated within the inferior collicular cortex. We found a withdrawal-related increase in the effectiveness of bicuculline to reduce the seizure threshold current within the inferior collicular cortex. Seizure sensitivity to collicular N -methyl- d -aspartic acid (NMDA) microinjection was decreased after 6 ethanol withdrawals, and increased after 10 withdrawals, but control liquid diet animals exhibited similar responses to collicular NMDA microinjection. Therefore, multiple withdrawals from ethanol alters the seizure sensitivity within the inferior collicular cortex. One possible contribution to this change is a local decrease in GABA inhibitory function.     

Age-related changes in GAD levels in the central auditory system of the rat

Changes in the levels of gamma-aminobutyric acid (GABA) are known to occur in different parts of the brain during aging. In our study we attempted to define the effect that aging has on glutamate decarboxylase (GAD), the key enzyme in the synthesis of GABA, in the central parts of the auditory system. Age-related changes in GAD65 and GAD67 levels were investigated using immunohistochemistry and Western blotting in the inferior colliculus (IC), the auditory cortex (AC) and the visual cortex in Long-Evans rats. The results show that aging is associated with a decrease in the numbers of GAD65- and 67-immunoreactive neurons and the optical density of their somas in both the IC and AC. Western blot analysis revealed a pronounced age-related decline in the levels of GAD65 and 67 proteins in both the IC and AC. For comparison, in the visual cortex the decrease in both proteins was less pronounced than in the IC and AC. A similar pattern of age-related changes was found in Fischer 344 rats, a strain that manifests a rapid loss of hearing function with aging. The observed age-related decline in the levels of GAD65 and 67 may contribute significantly to the deterioration of hearing function that accompanies aging in mammals, including man.