Moving beyond the concept of "primary forest" as a metric of forest environment quality.

The United Nations Food and Agriculture Organization (FAO) has been reporting country-level area in primary forests in its Global Forest Resource Assessment since 2005. The FAO definition of a primary forest (naturally regenerated forest of native species where there are no clearly visible indications of human activities and the ecological processes are not significantly disturbed) is generally accepted as authoritative and is being used in policy making. However, problems with this definition undermine our capacity to obtain globally coherent estimates. In addition, the current reporting on primary forests fails to consider the complementarily of non-primary forests toward the maintenance of ecosystem services. These issues undermine the appropriate tracking of changes in primary and non-primary forests, and the assessment of impacts of such changes on ecosystem services. We present the case for an operational reconsideration of the primary forest concept and discuss how alternatives or supplements might be developed.

Expression of 17beta-hydroxysteroid dehydrogenase type 2 and type 5 in breast cancer and adjacent non-malignant tissue: a correlation to clinicopathological parameters.

Estrogens play an important role in the development and progression of breast cancer. 17beta-Hydroxysteroid dehydrogenase (17beta-HSD) type 2 and type 5 are involved in sex steroid metabolism. 17beta-HSD type 2 converts estradiol to estrone while 17beta-HSD type 5 converts androstenedione to testosterone. Using immunocytochemistry, we have studied the expression of 17beta-HSD type 2 and type 5 in 50 specimens of breast carcinoma and adjacent non-malignant tissues. The results were correlated with the estrogen receptor alpha (ERalpha) and beta (ERbeta), progesterone receptor A (PRA) and B (PRB), androgen receptor and CDC47 and with the tumor stage, tumor size, nodal status and menopausal status. 17beta-HSD type 2 was expressed in 20% and 17beta-HSD type 5 in 56% of breast cancer specimens. In adjacent normal tissues, both enzymes were highly expressed in almost all the patients. No significant association could be found between the expression of 17beta-HSD type 2 and 17beta-HSD type 5 and between the expression of each enzyme and the clinicopathological parameters studied. The decrease in 17beta-HSD type 2 and 17beta-HSD type 5 expressions in breast cancer may play a predominant role in the development and/or progression of the cancer by modifying the intratumoral levels of estrogens and androgens.

A spatially structured network of inhibitory and excitatory connections directs impulse traffic within the lateral amygdala.

The lateral nucleus of the amygdala is the entry point of most sensory inputs into the amygdala. However, the way information is processed and distributed within the lateral nucleus still eludes us. To gain some insight into this issue, we have examined the spatial organization of excitatory and inhibitory connections in the lateral nucleus. To this end, we performed whole-cell recordings of principal lateral amygdala neurons and studied their responses to local pressure applications of glutamate in coronal and horizontal slices of the guinea-pig amygdala. In coronal sections, glutamate puffs performed at a distance from the recorded cells usually evoked inhibitory responses, except when the recorded neuron was adjacent to the external capsule, in which case excitatory responses could be evoked from ejection sites along the external capsule. In contrast, glutamate puffs evoked a mixture of excitatory and inhibitory responses in horizontal slices. Excitatory responses were generally evoked from stimulation sites located lateral to the recorded cell whereas inhibitory responses were commonly elicited from medial stimulation sites, irrespective of their rostrocaudal position. These findings confirm and extend previous tract-tracing studies where it was found that intrinsic connections within the lateral amygdala prevalently run in the dorsoventral and lateromedial directions. However, our results also reveal a hitherto unsuspected level of spatial heterogeneity in the intrinsic circuit of the lateral amygdala. The prevalence of excitatory responses in horizontal slices coupled to the ubiquity of inhibitory responses in coronal slices suggest that the lateral amygdala network is designed to allow associative interactions within the rostrocaudal plane while preventing runaway excitation locally.

Mechanisms underlying the formation of the amygdalar fear memory trace: A computational perspective.

Recent experimental and modeling studies on the lateral amygdala (LA) have implicated intrinsic excitability and competitive synaptic interactions among principal neurons (PNs) in the formation of auditory fear memories. The present modeling studies, conducted over an expanded range of intrinsic excitability in the network, revealed that only excitable PNs that received tone inputs participate in the competition. Strikingly, the number of model PNs integrated into the fear memory trace remained constant despite the much larger range considered, and model runs highlighted several conditioning-induced tone responsive characteristics of the various PN populations. Furthermore, these studies showed that although excitation was important, disynaptic inhibition among PNs is the dominant mechanism that keeps the number of plastic PNs stable despite large variations in the network's excitability. Finally, we found that the overall level of inhibition in the model network determines the number of projection cells integrated into the fear memory trace.

Altered responsiveness of BNST and amygdala neurons in trauma-induced anxiety.

A highly conserved network of brain structures regulates the expression of fear and anxiety in mammals. Many of these structures display abnormal activity levels in post-traumatic stress disorder (PTSD). However, some of them, like the bed nucleus of the stria terminalis (BNST) and amygdala, are comprised of several small sub-regions or nuclei that cannot be resolved with human neuroimaging techniques. Therefore, we used a well-characterized rat model of PTSD to compare neuronal properties in resilient vs PTSD-like rats using patch recordings obtained from different BNST and amygdala regions in vitro. In this model, a persistent state of extreme anxiety is induced in a subset of susceptible rats following predatory threat. Previous animal studies have revealed that the central amygdala (CeA) and BNST are differentially involved in the genesis of fear and anxiety-like states, respectively. Consistent with these earlier findings, we found that between resilient and PTSD-like rats were marked differences in the synaptic responsiveness of neurons in different sectors of BNST and CeA, but whose polarity was region specific. In light of prior data about the role of these regions, our results suggest that control of fear/anxiety expression is altered in PTSD-like rats such that the influence of CeA is minimized whereas that of BNST is enhanced. A model of the amygdalo-BNST interactions supporting the PTSD-like state is proposed.

Synaptic competition in the lateral amygdala and the stimulus specificity of conditioned fear: a biophysical modeling study.

Competitive synaptic interactions between principal neurons (PNs) with differing intrinsic excitability were recently shown to determine which dorsal lateral amygdala (LAd) neurons are recruited into a fear memory trace. Here, we explored the contribution of these competitive interactions in determining the stimulus specificity of conditioned fear associations. To this end, we used a realistic biophysical computational model of LAd that included multi-compartment conductance-based models of 800 PNs and 200 interneurons. To reproduce the continuum of spike frequency adaptation displayed by PNs, the model included three subtypes of PNs with high, intermediate, and low spike frequency adaptation. In addition, the model network integrated spatially differentiated patterns of excitatory and inhibitory connections within LA, dopaminergic and noradrenergic inputs, extrinsic thalamic and cortical tone afferents to simulate conditioned stimuli as well as shock inputs for the unconditioned stimulus. Last, glutamatergic synapses in the model could undergo activity-dependent plasticity. Our results suggest that plasticity at both excitatory (PN-PN) and di-synaptic inhibitory (PN-ITN and, particularly, ITN-PN) connections are major determinants of the synaptic competition governing the assignment of PNs to the memory trace. The model also revealed that training-induced potentiation of PN-PN synapses promotes, whereas that of ITN-PN synapses opposes, stimulus generalization. Indeed, suppressing plasticity of PN-PN synapses increased, whereas preventing plasticity of interneuronal synapses decreased the CS specificity of PN recruitment. Overall, our results indicate that the plasticity configuration imprinted in the network by synaptic competition ensures memory specificity. Given that anxiety disorders are characterized by tendency to generalize learned fear to safe stimuli or situations, understanding how plasticity of intrinsic LAd synapses regulates the specificity of learned fear is an important challenge for future experimental studies.

Bistable behavior of inhibitory neurons controlling impulse traffic through the amygdala: role of a slowly deinactivating K+ current.

The intercalated cell masses of the amygdala are clusters of GABAergic neurons located strategically to influence behavioral responsiveness. Indeed, they receive glutamatergic sensory inputs from the basolateral amygdaloid complex and generate feedforward inhibition in neurons of the central amygdala that mediate important components of fear responses. In the present study, using whole-cell recording methods in coronal slices of the guinea pig amygdala, we show that the activity of intercalated neurons is a function of their recent firing history because they express an unusual voltage-dependent K(+) conductance (termed I(SD) for slowly deinactivating). This conductance activates in the subthreshold regime, inactivates in response to suprathreshold depolarizations, and deinactivates very slowly upon return to rest. As a result, after bouts of suprathreshold activity, these cells enter a self-sustaining state of heightened excitability associated with an increased input resistance and a membrane depolarization. In turn, these changes increase the likelihood that ongoing synaptic activity will trigger orthodromic action potentials. However, because each orthodromic spike "renews" the inactivation of I(SD), intercalated cells can remain hyperexcitable for a long time and, via the central amygdaloid nucleus, exert a lasting influence on behavior.

Neuronal correlates of fear in the lateral amygdala: multiple extracellular recordings in conscious cats.

Much data implicates the amygdala in the expression and learning of fear. Yet, few studies have examined the neuronal correlates of fear in the amygdala. This study aimed to determine whether fear is correlated to particular activity patterns in the lateral amygdaloid (LA) nucleus. Cats, chronically implanted with multiple microelectrodes in the LA and a catheter in the femoral artery, learned that a series of tones interrupted by a period of silence (5 sec) preceded the administration of a footshock. During the silent period, their blood pressure increased, indicating that they anticipated the noxious stimulus. In parallel, the firing rate of LA neurons doubled, and the discharges of simultaneously recorded cells became more synchronized. Moreover, cross-correlation of focal LA waves revealed a significant increase in synchrony restricted to the theta band. In keeping with this, perievent histograms of neuronal discharges revealed rhythmic changes in the firing probability of LA neurons in relation to focal theta waves. Finally, the responsiveness of LA cells to the stimuli predicting the footshock (the tones) increased during the trials, whereas responses to unrelated stimuli (perirhinal shocks) remained stable. Thus, during the anticipation of noxious stimuli, a state here defined anthropomorphically as fear, the firing rate of LA neurons increases, and their discharges become more synchronized through a modulation at the theta frequency. The presence of theta oscillations in the LA might facilitate cooperative interactions between the amygdala and cortical areas involved in memory.

Infralimbic cortex activation increases c-Fos expression in intercalated neurons of the amygdala.

Recently, it was reported that stimulation of the infralimbic cortex produces a feedforward inhibition of central amygdala neurons. The interest of this observation comes from the fact that the central nucleus is the main output station of the amygdala for conditioned fear responses and evidence that the infralimbic cortex plays a critical role in the extinction of conditioned fear. However, the identity of the neurons mediating this infralimbic-evoked inhibition of the central nucleus remains unknown. Likely candidates are intercalated amygdala neurons. Indeed, these cells receive glutamatergic afferents from the infralimbic cortex, use GABA as a transmitter, and project to the central amygdala. Thus, the present study was undertaken to test whether, in adult rats, the infralimbic cortex can affect the activity of intercalated neurons. To this end, disinhibition of the infralimbic cortex was induced by local infusion of the non-competitive GABA-A receptor antagonist picrotoxin. Subsequently, neuronal activation was determined bilaterally within the amygdala using induction of the immediate early gene Fos. Infralimbic disinhibition produced a significant increase in the number of Fos-immunoreactive intercalated cells bilaterally whereas no change was detected in the central nucleus. In the basolateral amygdaloid complex, increases in the number of Fos-immunoreactive cells only reached significance in the contralateral lateral nucleus. These results suggest that glutamatergic inputs from the infralimbic cortex directly activate intercalated neurons. Thus, our findings raise the possibility that the infralimbic cortex inhibits conditioned fear via the excitation of intercalated cells and the consequent inhibition of central amygdala neurons.

Intra-amygdaloid projections of the lateral nucleus in the cat: PHA-L anterograde labeling combined with postembedding GABA and glutamate immunocytochemistry.

Research on the implication of the amygdala in classical fear conditioning suggests that the central amygdaloid nucleus is the output station of the amygdala for conditioned fear responses, while the lateral nucleus acts as the input nucleus, at least for auditory conditioned stimuli. However, the nature and locus of the plastic changes taking place between these two nuclei are unknown partly because the neurotransmitter(s) used by intra-amygdaloid projections of the lateral nucleus has not been identified. To address this issue in cats, anterograde tracing with Phaseolus vulgaris-leucoagglutinin (PHA-L) was combined with postembedding immunocytochemistry for gamma-aminobutyric acid (GABA) and glutamate. Two sectors can be recognized in the lateral nucleus of the cat: a shell located laterally along the external capsule, and a core. Iontophoretic injections of PHA-L in these two sectors revealed that they have nonoverlapping intra-amygdaloid targets with the exception of a common projection to the central lateral nucleus. The core projects mainly to itself and to the basomedial nucleus, whereas the shell contributes a massive projection to the basolateral nucleus. No projection of the lateral nucleus to the central medial nucleus was found. Electron microscopically, PHA-L-labeled axon terminals in the lateral, basomedial, basolateral, and central lateral nuclei as well as in the perirhinal and insular cortices formed asymmetric synapses (100%; n = 289) with dendritic spines (77-100%). Moreover, postembedding immunocytochemistry revealed that PHA-L-labeled axon terminals are immunoreactive for glutamate but not GABA. Since most amygdaloid projections to the brainstem originate in the central medial nucleus, these results suggest that intra-amygdaloid targets of the lateral nucleus are involved in the transmission of auditory conditioned stimuli to the central medial nucleus. Moreover, these findings imply that intra-amygdaloid projections of the lateral nucleus use glutamate but not GABA as a neurotransmitter.

GABAergic projection from the intercalated cell masses of the amygdala to the basal forebrain in cats.

The intercalated cell masses (ICMs) are dense clusters of small GABAergic cells interposed between the basolateral and centromedial nuclear groups of the amygdala. Until now, the ICMs have been largely ignored in anatomical studies of the amygdaloid complex. Thus, this study was undertaken to identify some of their targets by means of tract-tracing methods combined with immunohistochemical techniques. Wheat-germ agglutinin conjugated to horseradish peroxidase (WGA-HRP) was injected into numerous cortical areas and dorsal thalamic nuclei, in the anterior commissure and/or stria terminalis nuclei, and in the caudate nucleus, as well as into lateral and preoptic hypothalamic areas. Very few retrogradely labeled cells were seen in the ICMs following these injections. In contrast, massive retrograde labeling was found in the rostral groups of ICMs after WGA-HRP injections involving the substantia innominata and horizontal limb of the diagonal band. Furthermore, these retrogradely labeled intercalated cells were also GABA-immunoreactive. Results of iontophoretic injections of Phaseolus vulgaris-leucoagglutinin (PHA-L) in the rostral ICMs confirmed that they contribute a massive projection to the entire extent of the substantia innominata and horizontal limb of the diagonal band. Electron microscopic observations of ultrathin sections prepared for postembedding GABA or glutamate immunocytochemistry revealed that the ICM terminals labeled with PHA-L displayed GABA, but not glutamate immunoreactivity, and formed symmetric synapses with dendritic profiles. The present findings constitute the first direct demonstration of an amygdalofugal GABAergic projection to the basal forebrain. Considering that the basal forebrain contains a group of cholinergic and GABAergic neurons collectively projecting to the entire cortical mantle, this GABAergic projection of the ICMs could allow the amygdaloid complex to influence the activity of widespread cortical regions to which it is not directly connected, at least in the cat.

Thalamic collaterals of corticostriatal axons: their termination field and synaptic targets in cats.

Branched cortical projections to the thalamus and striatum were investigated in cats by injecting the retrograde-anterograde tracer biotinylated-dextran amine (BDA) into the caudate nucleus. These injections gave rise to plexuses of labeled fibers and varicosities in widespread thalamic territories. For instance, the lateroposterior nucleus and pulvinar (LP-PUL) mostly contained thick axons that contributed clusters of large-sized varicosities, each forming multiple asymmetric synapses, usually with vesicle-filled dendrites. In contrast, the intralaminar nuclei mostly contained thin axonal segments that emitted small en passant varicosities that formed single asymmetric synapses with spines. Because the caudate nucleus does not project to the thalamus, this labeling had to arise from a neuronal population with branching axons to both structures. Previous findings pointed to three possible sources: brainstem monoaminergic cells, intralaminar thalamic neurons, and corticostriatal cells. The first candidate could be ruled out because monoaminergic neurons contribute small-sized terminals that usually lack membrane specializations. The second possibility was discarded because retrograde tracer injections into the LP-PUL did not give rise to retrograde labeling in the intralaminar nuclear complex but to massive retrograde labeling in deep layers of cortical areas 5 and 7. Therefore, we concluded that the thalamic anterograde labeling originated from corticostriatal neurons, with axons branching to the thalamus. In keeping with this conclusion, Phaseolus vulgaris-leucoagglutinin (PHA-L) injections into cortical areas 5-7 labeled a group of thick corticothalamic fibers that ended in clusters of large boutons in the LP-PUL. These PHA-L-positive terminals were indistinguishable from those labeled after injections of BDA into the caudate nucleus, but they were easy to distinguish from the typical corticothalamic fibers. These findings indicate that the cerebral cortex could coordinate the activity of the striatum and the thalamus via a rich axonal network that collateralizes to both structures. The extent and synaptic organization of this branched projection impose a revision of the traditional scheme of thalamic connectivity.

Cat intraamygdaloid inhibitory network: ultrastructural organization of parvalbumin-immunoreactive elements.

Projection neurons of the basolateral (BL) amygdaloid complex are regulated by an intrinsic inhibitory network. To improve our understanding of this inhibitory circuit, we studied the synaptology of parvalbumin-immunopositive (PV+) elements as this calcium-binding protein is localized in a subpopulation of gamma-aminobutyric acid (GABA)-ergic interneurons. Two populations of PV+ cells were identified on the basis of soma shape (ovoid, type A vs. polygonal, type B). In the lateral and BL nuclei, the majority of boutons in contact with PV+ cells formed asymmetric synapses (types 1-3; 94%), whereas a minority (type 4, 6%) established symmetric synaptic contacts and resembled GABAergic terminals. In both nuclei, type B PV+ perikarya were more densely innervated than were type A neurons. However, the pattern of synaptic innervation of type B PV+ neurons differed in the two nuclei: in the lateral nucleus, they were almost exclusively innervated by a population of small, presumed excitatory terminals (type 1), whereas the four categories of terminals contributed more equally to their innervation in the BL nucleus. PV+ boutons belonged to a single category of terminals that was enriched with GABA and formed symmetric synapses mostly with the proximal part of PV neurons. The proportion of axosomatic synapses was significantly higher in the lateral nucleus than in the BL nucleus (33% vs. 18%). The reverse was true for the contacts with proximal dendrites (33% in the lateral nucleus vs. 46% in the BL nucleus). The remaining terminals formed synapses with distal dendrites (23-28%) and spines (8-12%). These results indicate that PV+ interneurons receive massive excitatory inputs and that PV+ terminals are strategically located to exert a powerful inhibitory control of amygdala neurons.

Basal forebrain cholinergic and noncholinergic projections to the thalamus and brainstem in cats and monkeys.

The projections of basal forebrain neurons to the thalamus and the brainstem were investigated in cats and primates by using retrograde transport techniques and choline acetyltransferase (ChAT) immunohistochemistry. In a first series of experiments, the lectin wheat germ-agglutinin conjugated with horseradish peroxidase (WGA-HRP) was injected into all major sensory, motor, intralaminar, and reticular (RE) thalamic nuclei of cats and into the mediodorsal (MD) and pulvinar-lateroposterior thalamic nuclei of macaque monkeys. In cats numerous neurons of the vertical and horizontal limbs of the diagonal band nucleus and the substantia innominata (SI), including its rostromedial portion termed the ventral pallidum (VP), were retrogradely labeled after WGA-HRP injections in the rostral pole of the RE complex, the MD, and anteroventral/anteromedial (AV/AM) thalamic nuclei. Fewer retrogradely labeled cells were observed in the same areas after injections in the ventromedial (VM) thalamic nucleus, and none or very few after other thalamic injections. After RE, MD, and AV/AM injections, 7-20% of all retrogradely labeled cells in the basal forebrain were also ChAT positive, while none of the retrogradely labeled neurons following VM injections displayed ChAT immunoreactivity. The basal forebrain projection to the MD nucleus was shown to arise principally from VP in both cats and macaque monkeys. In a second series of experiments performed in cats, injections of WGA-HRP in the brainstem peribrachial (PB) area comprising the pedunculopontine nucleus led to retrograde labeling of a moderate number of neurons in the lateral part of the VP, SI, and preoptic area (POA), only a few of which displayed ChAT immunoreactivity. In addition, a large number of retrogradely labeled cells were observed in the bed nuclei of the anterior commissure and stria terminalis after PB injections. In a third series of experiments, the use of the retrograde double-labeling method with fluorescent tracers in squirrel monkeys allowed us to identify a significant number of basal forebrain neurons sending axon collaterals to both the RE thalamic nucleus and PB brainstem area, while no double-labeled neurons were disclosed after injections confined to the ventral anterior/ventral lateral (VA/VL) thalamic nuclei and PB area or following injections in the cerebral cortex and PB area. Our findings reveal the existence of cholinergic and noncholinergic basal forebrain projections to the thalamus and the brainstem in both cats and macaque monkeys. We suggest that these projections may play a crucial role in the control of thalamic functions in mammals.

Differential innervation of parvalbumin-immunoreactive interneurons of the basolateral amygdaloid complex by cortical and intrinsic inputs.

In the basolateral (BL) amygdaloid complex, the excitability of projection cells is regulated by intrinsic inhibitory interneurons using gamma-aminobutyric acid (GABA) as a transmitter. A subset of these cells are labeled in a Golgi-like manner by Parvalbumin (PV) immunohistochemistry. Recently, we have shown that the overwhelming majority of axon terminals contacting these PV-immunoreactive neurons form asymmetric synapses. The present study was undertaken to identify the source(s) of these inputs. Since previous work had revealed that thalamic axons form very few synapses on BL interneurons (< 1%), we focused on cortical and intra-amygdaloid inputs. Iontophoretic injections of the anterograde tracers Phaseolus vulgaris-leucoagglutinin or biotinylated dextran amine were performed in various cortical fields in cats (perirhinal, entorhinal, pre/infralimbic cortices) and monkeys (orbitofrontal region) or in the BL amygdaloid nucleus in cats. These injections resulted in a large number of anterogradely labeled terminals forming asymmetric synapses in the BL complex. Following cortical injections, numerous anterogradely labeled terminals were found in the vicinity of PV-immunoreactive interneurons in the BL amygdala. However, only approximately 1% of these terminals formed synaptic contacts with PV-immunoreactive profiles. In contrast, as many as 11% of the terminals contributed by the intranuclear axon collaterals of BL projection cells established synapses with PV-immunoreactive elements. Since the axon terminals of PV-immunoreactive interneurons are enriched in GABA and they exclusively form symmetric synapses, these results suggest that PV-immunoreactive interneurons are predominantly involved in feedback inhibition in the BL amygdaloid complex.

Conscious and pre-conscious processes as seen from the standpoint of sleep-waking cycle neurophysiology.

The literature on state-dependent fluctuations in thalamocortical activities indicates that in electrophysiological terms, waking and paradoxical sleep are fundamentally identical states, with the provision that the handling of sensory information is altered in REM sleep. The central paradox of REM sleep, namely the apparent lack of cognitive responsiveness to sensory stimulation in spite of increased thalamocortical responsiveness to sensory stimuli, will lead us to hypothesize that the processing of sensory inputs in REM sleep is similar to that underlying preconscious processing of sensory inputs in the waking state. This will lead to a general discussion of the role of fast (approximately equal to 40 Hz) thalamocortical oscillations and temporal binding in sensory processing and conscious experience.

Distribution of GABA immunoreactivity in the amygdaloid complex of the cat.

This study describes the distribution of GABA immunoreactivity in the amygdaloid complex of cats. At the light microscopic level, immunopositive structures consisted of morphologically diverse somata and numerous small punctate elements. The latter accounted for most of the staining at low magnification and, at the electron microscopic level, were found to be axon terminals establishing symmetric synaptic contacts with a variety of postsynaptic profiles. Deep and superficial amygdaloid nuclei could be assigned to one of four groups according to (i) the intensity of immunolabeling they displayed, (ii) their density in reactive somata, and (iii) the size of the immunopositive somata they contained. Intercalated cell masses displayed the highest density of strongly immunoreactive cell bodies and presumed GABAergic terminals. However, electron microscope observations showed that intercalated somata were almost devoid of synaptic contacts. In contrast, central and medial nuclei were characterized by a low density of intensely immunoreactive somata and an elevated concentration for GABAergic terminals which contacted somatic and dendritic profiles. In addition, central and medial nuclei contained numerous neurons displaying low to moderate immunoreactivity. Superficial amygdaloid nuclei and nuclei of the basolateral complex displayed an intermediate density of immunoreactive somata and a low to moderate concentration of presumed terminals. Analysis of the distribution of soma areas within these nuclei revealed that the basolateral complex contains a distinct subpopulation of larger immunoreactive neurons. In light of recent electrophysiological findings, these results suggest that the intra-amygdaloid GABAergic system plays a major role in controlling the synaptic responsiveness and spontaneous activity of amygdaloid neurons.

The intercalated cell masses project to the central and medial nuclei of the amygdala in cats.

In order to study the efferent projections of the intercalated cell masses within the amygdaloid complex, iontophoretic injections of cholera toxin B subunit were performed in several amygdaloid nuclei of the centromedial and basolateral groups in cats. Analysis of the ensuing retrograde labeling revealed that the main intra-amygdaloid targets of the intercalated cell masses are the central and medial nuclei. Most intercalated neurons projecting to the medial nucleus were found in the larger, rostrally located intercalated cell masses. In contrast, the majority of intercalated cells projecting to the central medial and central lateral nuclei were found in the smaller, caudally located intercalated cell masses. In addition, evidence for weaker projections to the basolateral nucleus and other intercalated cell masses was obtained. In light of previous immunohistochemical results showing that GABAergic cells represent the main cell type in the intercalated cell masses, these results imply that the intercalated cell masses constitute an important source of GABAergic input to the centromedial complex. The significance of this finding lies in the fact that the intercalated cell masses are located at the interface between the basolateral nuclear group and the centromedial complex, the main route through which the amygdaloid complex can directly influence hypothalamic and brainstem centers involved in the elaboration of autonomic responses and species-specific emotional behaviors.

Bidirectional synaptic plasticity in intercalated amygdala neurons and the extinction of conditioned fear responses.

Classical fear conditioning is believed to result from potentiation of conditioned synaptic inputs in the basolateral amygdala. That is, the conditioned stimulus would excite more neurons in the central nucleus and, via their projections to the brainstem and hypothalamus, evoke fear responses. However, much data suggests that extinction of fear responses does not depend on the reversal of these changes but on a parallel NMDA-dependent learning that competes with the first one. Because they control impulse traffic from the basolateral amygdala to the central nucleus, GABAergic neurons of the intercalated cell masses are ideally located to implement this second learning. Consistent with this hypothesis, the present study shows that low- and high-frequency stimulation of basolateral afferents respectively induce long-term depression (LTD) and potentiation (LTP) of responses in intercalated cells. Moreover, induction of LTP and LTD is prevented by application of an NMDA antagonist. To determine how these activity-dependent changes are expressed, we tested whether LTD and LTP induction are associated with modifications in paired-pulse facilitation, an index of transmitter release probability. Only LTP induction was associated with a change in paired-pulse facilitation. Depotentiation of previously potentiated synapses did not revert the modification in paired pulse facilitation, suggesting that LTP is associated with presynaptic alterations, but that LTD and depotentiation depend on postsynaptic changes. Taken together, our results suggest that basolateral synapses onto intercalated neurons can express NMDA-dependent LTP and LTD, consistent with the possibility that intercalated neurons are a critical locus of plasticity for the extinction of conditioned fear responses. Ultimately, these plastic events may prevent conditioned amygdala responses from exciting neurons of the central nucleus, and thus from evoking conditioned fear responses.

Of dreaming and wakefulness.

Following a set of studies concerning the intrinsic electrophysiology of mammalian central neurons in relation to global brain function, we reach the following conclusions: (i) the main difference between wakefulness and paradoxical sleep lies in the weight given to sensory afferents in cognitive images; (ii) otherwise, wakefulness and paradoxical sleep are fundamentally equivalent brain states probably subserved by an intrinsic thalamo-cortical loop. From this assumption, we conclude that wakefulness is an intrinsic functional realm, modulated by sensory parameters. In support of this hypothesis, we review morphological studies of the thalamocortical system, which indicate that only a minor part of its connectivity is devoted to the transfer of direct sensory input. Rather, most of the connectivity is geared to the generation of internal functional modes, which may, in principle, operate in the presence or absence of sensory activation. These considerations lead us to challenge the traditional Jamesian view of brain function according to which consciousness is generated as an exclusive by-product of sensory input. Instead, we argue that consciousness is fundamentally a closed-loop property, in which the ability of cells to be intrinsically active plays a central role. We further discuss the importance of spatial and temporal mapping in the elaboration of cognitive and perceptual constructs.