Fabio Bordi

Somatosensory and auditory convergence in the lateral nucleus of the amygdala

Previous studies have shown that the lateral nucleus of the amygdala (AL) is essential in auditory fear conditioning and that neurons in the AL respond to auditory stimuli. The goals of the present study were to determine whether neurons in the AL are also responsive to somatosensory stimuli and, if so, whether single neurons in the AL respond to both auditory and somatosensory stimulation. Single-unit activity was recorded in the AL in anesthetized rats during the presentation of acoustic (clicks) and somatosensory (footshock) stimuli. Neurons in the dorsal subdivision of the AL responded to both somatosensory and auditory stimuli, whereas neurons in the ventrolateral AL responded only to somatosensory stimuli and neurons in the ventromedial AL did not respond to either stimuli. These findings indicate that the dorsal AL is a site of auditory and somatosensory convergence and may therefore be a locus of convergence of conditioned and unconditioned stimuli in auditory fear conditioning.

Response properties of single units in areas of rat auditory thalamus that project to the amygdala - I. Acoustic discharge patterns and frequency receptive fields

Projections from the auditory thalamus to the amygdala have been implicated in the processing of the emotional signficance of auditory stimuli. In order to further our understanding of the contribution of thalamoamygdala projections to auditory emotional processing, acoustic response properties of single neurons were examined in the auditory thalamus of chloral hydrate-anesthetized rats. The emphasis was on the medial division of the medial geniculate body (MGm), the suprageniculate nucleus (SG), and the posterior intralaminar nucleus (PIN), thalamic areas that receive inputs from the inferior colliculus and project to the lateral nucleus of the amygdala (AL). For comparison, recordings were also made from the specific thalamocortical relay nucleus, the ventral division of the medial geniculate body (MGv). Responses latencies were not statistically different in MGv, MGm, PIN, and SG, but were longer in the posterior thalamic region (PO). Overall, frequency tuning functions were narrower in MGv than in the other areas but many cells in MGm were as narrowly tuned as cells in MGv. There was some organization of MGv, with low frequencies represented dorsally and high frequencies ventrally. A similar but considerably weaker organization was observed in MGm. While the full range of frequencies tested (1–30 kHz) was represented in MGv, cells in MGm, PIN, and SG tended to respond best to higher frequencies (16–30 kHz). Thresholds were higher in PIN than in MGv (other areas did not differ from MGv). Nevertheless, across the various areas, the breadth of tuning was inversely related to threshold, such that more narrowly tuned cells tended to have lower thresholds. Many of the response properties observed in MGm, PIN, and SG correspond with properties found in AL neurons and thus add support to the notion that auditory responses in AL reflect thalamoamygdala transmission.

Response properties of single units in areas of rat auditory thalamus that project to the amygdala - II. Cells receiving convergent auditory and somatosensory inputs and cells antidromically activated by amygdala stimulation

The purpose of this study was to further our understanding of the contribution of auditory thalamoamygdala projections to conditioned emotional memories formed when auditory and noxious somatosensory stimuli are associated. Single unit activity was recorded in the acoustic thalamus of chloral hydrate-anesthetized rats in response to auditory (white noise, clicks, tones) and somatosensory (foot-shock) stimulation. The thalamic areas focused on were the the medial division of the medial geniculate body (MGm), the suprageniculate nucleus (SG), and the posterior intralaminar nucleus (PIN), thalamic areas that receive inputs from both the inferior colliculus and the spinal cord and that project to the lateral nucleus of the amygdala (AL). For comparison, recordings were also made from the specific thalamocortical relay nucleus, the ventral division of the medial geniculate body (MGv), which receives projections from the inferior colliculus but not from the spinal cord. Auditory but not somatosensory responses were recorded from MGv, while both auditory and somatosensory responses were frequently found in MGm, PIN, and SG. In these areas, convergent auditory and somatosensory responses were more frequently found rostrally than caudally. Within a thalamic subregion, the acoustic response properties of the convergence cells were not different from the response properties of unimodal auditory cells. Some cells that responded to somatosensory but not auditory stimuli showed a potentiated response when tested with simultaneous presentation of auditory and somatosensory stimuli. In some studies, thalamic cells that project to the amygdala were antidromically activated by stimulation of the AL. Consistent with anatomical tracing results, antidromically activated cells were found in MGm, PIN, and SG, but not in MGv. Antidromically activated cells were more likely to respond to auditory stimuli than to somatosensory stimuli, but unimodal somatosensory and convergence cells were also found. These findings, which provide the first characterization of acoustic response properties of multimodal cells in the auditory thalamus and of cells in the auditory thalamus that project to amygdala, suggest insights into the emotional functions of the thalamoamygdala pathway.

Single-unit activity in the lateral nucleus of the amygdala and overlying areas of the striatum in freely behaving rats: rates, discharge patterns, and responses to acoustic stimuli.

Acoustic responses of single units were examined in awake, freely behaving rats in the lateral nucleus of the amygdala (AL). Recordings were made from a movable bundle of 9 microwires. Most cells had very low rates of spontaneous activity (about 3 spikes/s average). Firing rates increased during sleep states. Short-latency auditory responses (12-25 ms) were found in the dorsal subnucleus (ALd) of the AL. Cells in the ALd most typically responded in a sustained fashion. Some of the cells in the ALd showed preferences for high frequencies, tone bursts, or frequency-modulated stimuli with center frequencies above 12 kHz. Response latencies were considerably longer in other areas of the amygdala. Our results corroborate the main findings of a previous study (F. Bordi & J. LeDoux, 1992) that examined the acoustic response properties of single cells in the AL in anesthetized rats. Together the findings from awake and anesthetized rats provide the most precise information about sensory processing in amygdala neurons available to date.

The Glycine Antagonist GV150526 Protects Somatosensory Evoked Potentials and Reduces the Infarct Area in the MCAo Model of Focal Ischemia in the Rat

The neuroprotective activity of the novel, selective glycine antagonist GV150526 was assessed in the middle artery occlusion (MCAo) model of focal ischemia. Postischemia administration of GV150526 (3 mg/kg i.v.) up to 6 h post-MCAo resulted in a significant reduction of the infarct volume measured histologically 24 h later. The neuronal protection by GV150526 was accompanied by functionally significant protection determined by somatosensory evoked potential (SEP) responses recorded from the primary somatosensory cortex of rats under urethane anesthesia. Experimental occlusion of the MCA 7 days prior to electrophysiological testing induced a clear reduction in the SEP amplitude. GV150526 (3mg/kg, i.v.) was able to protect SEP responses recorded from the hindpaw cortical field in two groups of animals treated either 1 (n = 9) or 6 h (n = 10) post-MCAo. SEP responses recorded from the forepaw cortical field, an area closer to the core of the ischemic damage, were significantly protected only in the group treated 1 h post-MCAo. Histological evaluation of the rat brain regions showed a correlated decrease in the ischemic area of GV150526-treated groups. The volumes of the ischemic brains of both GV150526 groups were statistically different from the MCAo group (P < 0.05). These findings demonstrate that GV150526 is able to prevent the ischemic damage assessed histologically and affect the functional correlates of the ischemia evaluated by the electrophysiological SEP measurements.

Enhanced behavioral stereotypies elicited by intrastriatal injection of D1 and D2 dopamine agonists in intact rats

Five components of behavior elicited by dopamine (DA) agonists (locomotor hyperactivity, sniffing, oral activity, grooming and paw nibbling) were evaluated after bilateral infusion of the selective D1 agonist fenoldopam (SKF 82526; 2.5-10 micrograms), the selective D2 agonist quinpirole (LY 171555; 5-40 micrograms) and the muscarinic cholinergic antagonist scopolamine (5-20 micrograms) into the ventral striatum of awake, unrestrained rats. Simultaneous bilateral infusion of various dose combinations of fenoldopam (2.5-10 micrograms) and quinpirole (5-20 micrograms) elicited dramatic increases in stereotyped behaviors relative to the effects produced by corresponding doses of each drug alone. Stereotyped sniffing and paw nibbling (self-directed oral activity) were markedly enhanced, whereas conventional oral behaviors (licking, chewing and/or biting) were either slightly or not at all increased. These potentiated responses were reduced or blocked by concomitant infusion of either the selective D1 antagonist SCH 23390 (1 and 5 micrograms) or the selective D2 antagonist sulpiride (0.15 microgram). Scopolamine (10 micrograms) only slightly increased the effects of quinpirole (5 micrograms) on both sniffing and oral behaviors, whereas it dramatically potentiated the effects of fenoldopam (2.5 micrograms) on oral activity; sniffing was only slightly increased. The effects of both drug combinations were almost completely antagonized by infusion of either SCH 23390 (1 microgram) or sulpiride (0.1 microgram). The results demonstrate that the synergistic effects of co-activation of D1 and D2 receptors observed after systemic administration are mediated at least in part by an interaction at the level of the striatum. Differences and similarities between the behaviors expressed after various treatments are discussed.

Stereotypies elicited by injection of N-propylnorapomorphine into striatal subregions and nucleus accumbens

Injection of the dopamine (DA) agonist R-(-)-N-n-propylnorapomorphine (NPA; 5-40 micrograms) into anterior ventral striatal sites (either lateral (VL) or medial (VM) elicited dose-dependent oral and sniffing stereotypies of rapid onset, long duration and high intensity. In contrast, injection into anterior dorsolateral (DL) or posterior ventral (lateral (PL) or medial (PM] sites produced little oral and moderate sniffing behavior of slower onset, shorter duration and low intensity. Injection into the dorsomedial (DM) striatum produced intermediate effects. Intra-accumbens NPA elicited weak oral activity and moderate sniffing which was similar in onset, duration and intensity to the least sensitive striatal sites (DL, PM and PL). In other experiments, DA receptors were inactivated with the irreversible blocking agent N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ; 6 mg/kg) and behavioral recovery was monitored by challenge with 20 micrograms NPA into the VL or the nucleus accumbens (NA) at various times after EEDQ. Sniffing behavior recovered rapidly (normal by day 4 in both regions), whereas oral activity required 8 (NA) and 12 days (VL) to return to control levels. The results are discussed in terms of a possible topographic distribution of behavior in the striatum. Alternatively, heterogeneity of DA receptor density may account for these findings.

Involvement of mGluR5 on acute nociceptive transmission

The effect of the mGluR(5) antagonist, MPEP (2-Methyl-6-(phenylethynyl)-pyridine), and of the mGluR(1) antagonist, AIDA((RS)-1-Aminoindan-1,5-dicarboxylic acid), were examined on nociceptive neurons in the ventroposterolateral (VPL) nucleus of the thalamus in response to pressure stimuli to the contralateral hindpaw of rats under urethane anesthesia. Intravenous (i.v.) injection of MPEP (0.1, 1, and 10 mg/kg) blocked responses to noxious stimulation in a dose-dependent and reversible manner. AIDA (3 and 15 mg/kg, i.v.), in contrast, had no effect on these cells. MPEP action was selective to noxious stimulation because even when tested at the highest dose (10 mg/kg, i.v.) it did not alter the responses of non-nociceptive neurons to brush stimulation. To investigate the site of action of MPEP, intra-thalamic injections were made during electrophysiological recordings. Using this method, the mGluR(5) antagonist did not affect nociceptive responses, suggesting that thalamic receptors were not involved in this action. On the other hand, the NMDA thalamic receptors seem to be involved because the NMDA receptor antagonist, MK801, successfully blocked responses to noxious pressure stimulation following intra-thalamic injections. In the spinal cord in vitro model, MPEP (30 microM, 60 min) was also able to attenuate ventral root potentials after single shock electrical stimulation of the dorsal root and inhibit wind-up response evoked by repetitive stimulation. Taken together, these findings suggest that blockade of the mGluR(5), but not mGluR(1) decreases nociceptive transmission in the thalamus and that these effects may be mediated by spinal cord receptors.

Enhancement by the D1 dopamine agonist SKF 38393 of specific components of stereotypy elicited by the D2 agonists LY 171555 and RU 24213

Dose-effect curves were obtained for five specific components of stereotyped behavior (locomotor hyperactivity, rearing, sniffing, grooming and oral activity) elicited by the nonselective dopamine agonists N-propylnorapomorphine (NPA; 0.001-4.5 mg/kg) and apomorphine (APO; 0.3-10.0 mg/kg), the selective D2 agonists LY 171555 (0.1-18.0 mg/kg) and RU 24213 (0.7-20.0 mg/kg) and the selective D1 agonist SKF 38393 (0.6-40.0 mg/kg) in male Sprague-Dawley rats. All the agonists except SKF 38393 elicited dose-dependent sniffing. Both NPA and APO produced robust oral activity at high doses, with concomitant reductions in other behaviors. Neither RU 24213 nor SKF 38393 elicited dose-dependent oral behavior, whereas LY 171555 induced a maximal level of oral activity which was much less intense than that produced by NPA or APO. SKF 38393 alone induced only dose-dependent grooming. Co-treatment of Sprague-Dawley rats with SKF 38393 (5-40 mg/kg) and either LY 171555 (0.1-1.6 mg/kg) or RU 24213 (6.7 mg/kg) produced significant enhancement of sniffing but not of oral activity. Significant enhancement of both behavioral components was observed in male Wistar rats. Differences in specific components of behavioral response on treatment with combinations of D1 and D2 agonists may reflect use of animal strains with different ratios of D1/D2 innervation and/or different behavioral criteria.

Regulation of synaptic plasticity by mGluR1 studied in vivo in mGluR1 mutant mice.

The role of the metabotropic glutamate receptor 1 (mGluR1) in synaptic plasticity was investigated in vivo in the intact hippocampus of mutant mice lacking this receptor. In a previous study we showed reduced long-term potentiation (LTP) in the dentate gyrus of mGluR1 -/- mice in vivo, but not when LTP was studied in a slice preparation. A possible explanation of this difference is that dentate neurons receive more inhibitory synaptic drive in vivo than in slice preparation where many inhibitory axon collaterals are lost. We report here that another form of synaptic plasticity, paired-pulse depression of the population spike, is also abnormal in the dentate gyrus of mGluR1-deficient mice when tested in vivo. In wild-type mice, stimulation of the medial perforant path produced paired-pulse depression of inter-pulse intervals (IPIs) up to 30 ms. Mutant mGluR1, on the other hand, showed a significantly longer IPI depression, up to 50 ms. Paired-pulse depression results from the activation of inhibitory interneurons. The GABA(B) agonist baclofen, acting presynaptically on the GABA interneurons, attenuated paired-pulse depression and allowed for a normal and stable LTP in mGluR1 mutant mice. These findings suggest an indirect role for mGluR1 in synaptic plasticity via a regulation of GABA inhibition.