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Synaptic inputs of class III and class V interneurons in the cat pulvinar nucleus: differential integration of RS and RL inputs.

We previously reported that two types of visual thalamic interneurons (class III and class V) could be distinguished by the presence or absence of the enzyme brain nitric oxide synthase (bNOS; Bickford et al., 1999). In the present study, we found that further immunocytochemical characterization can be used to isolate class V and class III neurons: class V neurons express GABA and bNOS but not the calcium binding protein calbindin, whereas class III neurons express GABA and calbindin, but not bNOS. By pairing staining for bNOS or calbindin with staining for GABA, we were able to distinguish class III and class V neurons, thereby allowing us to characterize the synaptic inputs to these two interneuron populations within the cat pulvinar nucleus. We found class III and class V interneurons participate in distinctly different circuitries. Class III cells receive the majority of their input from large terminals with round vesicles (RL profiles, that presumably originate from cortical layer V) while class V cells receive all of their input from small terminals with round vesicles (RS profiles, that presumably originate from cortical layer VI). These two types of interneurons also target different regions of the dendritic arbors of thalamocortical cells; class III cells contact large caliber (presumably proximal) thalamocortical dendrites within glomeruli, while class V cells contact the small caliber (presumably distal) thalamocortical cell dendrites within the extraglomerular neuropil. Thus, a dual inhibitory system exists within the visual thalamus that integrates different types of corticothalamic input and targets distinct regions of thalamocortical cell dendritic arbors.

Animals↗

Electrophysiological and theoretical analysis of depolarization-dependent outward currents in the dendritic membrane of an identified nonspiking interneuron in crayfish.

Depolarization-dependent outward currents were analyzed using the single-electrode voltage clamp technique in the dendritic membrane of an identified nonspiking interneuron (LDS interneuron) in situ in the terminal abdominal ganglion of crayfish. When the membrane was depolarized by more than 20 mV from the resting potential (65.0 +/- 5.7 mV), a transient outward current was observed to be followed by a sustained outward current. Pharmacological experiments revealed that these outward currents were composed of 3 distinct components. A sustained component (I(s)) was activated slowly (half rise time > 5 msec) and blocked by 20 mM TEA. A transient component (I(t1)) that was activated and inactivated very rapidly (peak time < 2.5 msec, half decay time < 1.2 msec) was also blocked by 20 mM TEA. Another transient component (I(t2)) was blocked by 100 microM 4AP, activated rapidly (peak time < 10.0 msec) and inactivated slowly (half decay time > 131.8 msec). Two-step pulse experiments have revealed that both sustained and transient components are not inactivated at the resting potential: the half-maximal inactivation was attained at -21.0 mV in I(t1), and -38.0 mV in I(t2). I(s) showed no noticeable inactivation. When the membrane was initially held at the resting potential level and clamped to varying potential levels, the half-maximal activation was attained at -36.0 mV in I(s), -31.0 mV in I(t1) and -40.0 mV in I(t2). The activation and inactivation time constants were both voltage dependent. A mathematical model of the LDS interneuron was constructed based on the present electrophysiological records to simulate the dynamic interaction of outward currents during membrane depolarization. The results suggest that those membrane conductances found in this study underlie the outward rectification of the interneuron membrane as well as depolarization-dependent shaping of the excitatory synaptic potential observed in current-clamp experiments.

4-Aminopyridine↗

Analysis of the dynamics of interneuronal functional connections during conditioned reflex activity.

The dynamics of interneuronal functional connections were studied in the prefrontal cortex of dogs performing a task consisting of unforeseen remodeling of movement conditioned reflexes. An original method was used to find and classify the temporal patterns of linked spikes coming from several simultaneously recorded neurons. This procedure showed that in 33 pairs of neurons (87% of the total number of pairs showing interneuronal functional connections), parts of the conditioned reflex program were associated with behaviorally significant changes in the functional relationship between the neurons. In different behavioral situations, linked activation of a given pair of cells was restricted to different stages of the performance of the conditioned reflex task. In 17 pairs of neurons, the periods of "switching of interneuronal functional connections," i.e., the intervals in which linked spikes were absent during all stages of task performance, were seen only in a particular behavioral context different from the situation in which these same cells generated linked discharges. The specific characteristics of different methods of analyzing the dynamics of interneuronal functional connections in conditions of a dynamic behavioral context and multifactorial determination of conditioned reflex activity are discussed.

Animals↗

Afferent-specific innervation of two distinct AMPA receptor subtypes on single hippocampal interneurons.

Using the polyamine toxin philanthotoxin, which selectively blocks calcium-permeable AMPA receptors, we show that synaptic transmission onto single hippocampal interneurons occurs by afferent-specific activation of philanthotoxin-sensitive and -insensitive AMPA receptors. Calcium-permeable AMPA receptors are found exclusively at synapses from mossy fibers. In contrast, synaptic responses evoked by stimulation of CA3 pyramidal neurons are mediated by calcium-impermeable AMPA receptors. Both pathways converge onto single interneurons and can be discriminated with Group II mGluR agonists. Thus, single interneurons target AMPA receptors of different subunit composition to specific postsynaptic sites, providing a mechanism to increase the synapse-specific computational properties of hippocampal interneurons.

Afferent Pathways↗

Calcium-permeable AMPA receptors mediate long-term potentiation in interneurons in the amygdala.

Fear conditioning is a paradigm that has been used as a model for emotional learning in animals. The cellular correlate of fear conditioning is thought to be associative N-methyl-D-aspartate (NMDA) receptor-dependent synaptic plasticity within the amygdala. Here we show that glutamatergic synaptic transmission to inhibitory interneurons in the basolateral amygdala is mediated solely by alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors. In contrast to AMPA receptors at inputs to pyramidal neurons, these receptors have an inwardly rectifying current-voltage relationship, indicative of a high permeability to calcium. Tetanic stimulation of inputs to interneurons caused an immediate and sustained increase in the efficacy of these synapses. This potentiation required a rise in postsynaptic calcium, but was independent of NMDA receptor activation. The potentiation of excitatory inputs to interneurons was reflected as an increase in the amplitude of the GABA(A)-mediated inhibitory synaptic current in pyramidal neurons. These results demonstrate that excitatory synapses onto interneurons within a fear conditioning circuit show NMDA-receptor independent long-term potentiation. This plasticity might underlie the increased synchronization of activity between neurons in the basolateral amygdala after fear conditioning.

Amygdala↗

Interneurons unbound.

Local-circuit, gamma-aminobutyric acid-releasing inhibitory interneurons of the hippocampus and cortex have traditionally been considered as the regulators of principal neuron activity--the yin to the excitatory yang. Recent evidence indicates that, in addition to that role, their network connectivity and the properties of their intrinsic voltage-gated currents are finely tuned to permit inhibitory interneurons to generate and control the rhythmic output of large populations of both principal cells and other populations of inhibitory interneurons. This review brings together recently described properties and emerging principles of interneuron function that indicate a much more complex role for these cells than just providers of inhibition.

Animals↗

Synchronization of neuronal activity in hippocampus by individual GABAergic interneurons.

SYNCHRONIZATION of neuronal activity is fundamental in the operation of cortical networks. With respect to an ongoing synchronized oscillation, the precise timing of action potentials is an attractive candidate mechanism for information coding. Networks of inhibitory interneurons have been proposed to have a role in entraining cortical, synchronized 40-Hz activity. Here we demonstrate that individual GABAergic interneurons can effectively phase spontaneous firing and subthreshold oscillations in hippocampal pyramidal cells at 0 frequencies (4-7 Hz). The efficiency of this entrainment is due to interaction of GABAA-receptor-mediated hyperpolarizing synaptic events with intrinsic oscillatory mechanisms tuned to this frequency range in pyramidal cells. Moreover, this GABAergic mechanism is sufficient to synchronize the firing of pyramidal cells. Thus, owing to the divergence of each GABAergic interneuron, more than a thousand pyramidal cells may share a common temporal reference established by an individual interneuron.

Action Potentials↗

Hebbian LTP in feed-forward inhibitory interneurons and the temporal fidelity of input discrimination.

Cortical information processing requires a delicate balance of excitatory and inhibitory signaling. How is this balance preserved during hippocampal memory encoding, which involves NMDA receptor-dependent long term potentiation (LTP)? This form of LTP occurs at synapses between pyramidal neurons but has not been detected in feed-forward inhibitory interneurons. We show that paired pre- and postsynaptic activity evokes pathway-specific LTP in half of rat stratum radiatum interneurons if cytoplasmic integrity is preserved. LTP occurs in aspiny feed-forward interneurons and propagates to pyramidal neurons as an enhancement of disynaptic inhibition. We also show that when LTP is restricted to synapses on pyramidal neurons, the temporal fidelity of synaptic integration and action potential generation in pyramidal cells is compromised. However, when LTP also occurs at synapses on feed-forward interneurons, temporal fidelity is preserved. We propose that Hebbian LTP at synapses driving disynaptic inhibition is necessary to maintain information processing without degradation during memory encoding.

Animals↗

RGS4-dependent attenuation of M4 autoreceptor function in striatal cholinergic interneurons following dopamine depletion.

Parkinson disease is a neurodegenerative disorder whose symptoms are caused by the loss of dopaminergic neurons innervating the striatum. As striatal dopamine levels fall, striatal acetylcholine release rises, exacerbating motor symptoms. This adaptation is commonly attributed to the loss of interneuronal regulation by inhibitory D(2) dopamine receptors. Our results point to a completely different, new mechanism. After striatal dopamine depletion, D(2) dopamine receptor modulation of calcium (Ca(2+)) channels controlling vesicular acetylcholine release in interneurons was unchanged, but M(4) muscarinic autoreceptor coupling to these same channels was markedly attenuated. This adaptation was attributable to the upregulation of RGS4-an autoreceptor-associated, GTPase-accelerating protein. This specific signaling adaptation extended to a broader loss of autoreceptor control of interneuron spiking. These observations suggest that RGS4-dependent attenuation of interneuronal autoreceptor signaling is a major factor in the elevation of striatal acetylcholine release in Parkinson disease.

Acetylcholine↗

The origin and specification of cortical interneurons.

GABA-containing interneurons are crucial to both the development and function of the cerebral cortex. Unlike cortical projection neurons, which have a relatively conserved set of characteristics, interneurons include multiple phenotypes that vary on morphological, physiological and neurochemical axes. This diversity, and the relatively late, context-dependent maturation of defining features, has challenged efforts to uncover the transcriptional control of cortical interneuron development. Here, we discuss recent data that are beginning to illuminate the origins and specification of distinct subgroups of cortical interneurons.

Animals↗

Serotoninergic and noradrenergic axonal contacts associated with premotor interneurons in spinal pathways from group II muscle afferents.

We investigated the possibility that monoaminergic axons make contacts with spinal interneurons which project to motor nuclei and are monosynaptically activated by group II muscle afferents. Interneurons in midlumbar spinal segments of adult cats were characterized electrophysiologically and intracellularly labelled with tetramethylrhodamine dextran. Serotoninergic and noradrenergic axons were identified with immunofluorescence in sections containing labelled cells. Contacts between monoaminergic axons and interneurons were investigated with three-colour confocal laser scanning microscopy and analysed with a computer reconstruction program. Cell bodies and dendritic trees of five cells were reconstructed and putative contacts were plotted. The average number of contacts formed by serotoninergic axons was 140 and the average number of noradrenergic contacts was 38. The majority (95%) of contacts were formed with dendrites; these were distributed over the entire dendritic tree, even on the most distal branches. These findings provide a morphological basis for the modulatory actions of monoamines on premotor spinal interneurons in pathways from group II muscle afferents.

Animals↗

Subcellular localization of m2 muscarinic receptors in GABAergic interneurons of the olfactory bulb.

We analysed the ultrastructural distribution of the m2 muscarinic receptor (m2R) in the rat olfactory bulb (OB) using immunohistochemical techniques and light and electron microscopy. m2R was differentially distributed within the cellular compartments of gamma-aminobutyric acid (GABA)ergic bulbar interneurons. It is located in the gemmules of granule cells and in the synaptic loci of the interneurons of the external plexiform layer, suggesting that m2R activation could modulate the release of GABA from these interneurons onto principal cells by a presynaptic mechanism. By contrast, the receptor appears in the somata and dendritic trunks of second-order short-axon interneurons located in the inframitral layers, suggesting that postsynaptic muscarinic activation in these cells could elicit the inhibition of granule cells, leading to a disinhibition of principal cells. We also detail the anatomical substrate for a new putative muscarinic modulation that has not been previously described, and that could influence the reception of sensory information within the olfactory glomeruli. m2R appears in a subset of GABAergic/dopaminergic juxtaglomerular cells innervated by olfactory axons but is absent in juxtaglomerular cells that do not receive sensory inputs. This finding suggests that m2R activation could modify, through dopaminergic local circuits, the strength of olfactory nerve inputs onto principal cells. Activation of the muscarinic receptor may modulate the olfactory information encoding within olfactory glomeruli and may facilitate the bulbar transmission to superior centres influencing the GABA release by presynaptic and postsynaptic mechanisms. Taken together, our data provide the neuroanatomical basis for a complex action of m2R at different levels in the mammalian OB.

Acetylcholine↗

Interneurons are the local targets of hippocampal inhibitory cells which project to the medial septum.

A subset of GABAergic neurons projecting to the medial septum has long been described in the hippocampus. However, the lack of information about their local connectivity pattern or their correspondence with any of the well-established hippocampal interneuron types has hampered the understanding of their functional role. Retrograde tracing combined with immunostaining for neurochemical markers in the adult rat hippocampus showed that nearly all hippocampo-septal (HS) neurons express somatostatin (>95%) and, in the hilus and CA3 stratum lucidum, many contain calretinin (>45%). In contrast, in stratum oriens of the CA1 and CA3 subfields, the majority of HS neurons contain somatostatin (>86%) and calbindin (>73%), but not calretinin. Because somatostatin-positive hippocampal interneurons have been most extensively characterized in the stratum oriens of CA1, we focused our further analysis on HS cells found in this region. In 18-20-day-old rats, intracellularly filled CA1-HS cells had extensive local axon collaterals crossing subfield boundaries and innervating the CA3 region and the dentate gyrus. Electron microscopic analysis provided evidence that the axon terminals of CA1-HS cells form symmetrical synapses selectively on GABAergic interneurons, both locally and in the CA3 region. In addition, double retrograde labelling experiments revealed that many CA1-HS neurons of the dorsal hippocampus also have collateral projections to the ventral hippocampus. Thus, CA1-HS cells innervate inhibitory interneurons locally and in remote hippocampal regions, in addition to targeting mostly GABAergic neurons in the medial septum. This dual projection with striking target selectivity for GABAergic neurons may be ideally suited to synchronize neuronal activity along the septo-hippocampal axis.

Animals↗

High level of mGluR7 in the presynaptic active zones of select populations of GABAergic terminals innervating interneurons in the rat hippocampus.

The release of neurotransmitters is modulated by presynaptic metabotropic glutamate receptors (mGluRs), which show a highly selective expression and subcellular location in glutamatergic terminals in the hippocampus. Using immunocytochemistry, we investigated whether one of the receptors, mGluR7, whose level of expression is governed by the postsynaptic target, was present in GABAergic terminals and whether such terminals targeted particular cells. A total of 165 interneuron dendritic profiles receiving 466 synapses (82% mGluR7a-positive) were analysed. The presynaptic active zones of most GAD-(77%) or GABA-positive (94%) synaptic boutons on interneurons innervated by mGluR7a-enriched glutamatergic terminals (mGluR7a-decorated) were immunopositive for mGluR7a. GABAergic terminals on pyramidal cells and most other interneurons in str. oriens were mGluR7a-immunonegative. The mGluR7a-decorated cells were mostly somatostatin- and mGluR1alpha-immunopositive neurons in str. oriens and the alveus. Their GABAergic input mainly originated from VIP-positive terminals, 90% of which expressed high levels of mGluR7a in the presynaptic active zone. Parvalbumin-positive synaptic terminals were rare on mGluR7a-decorated cells, but on these neurons 73% of them were mGluR7a-immunopositive. Some type II synapses innervating interneurons were immunopositive for mGluR7b, as were some type I synapses. Because not all target cells of VIP-positive neurons are known it has not been possible to determine whether mGluR7 is expressed in a target-cell-specific manner in the terminals of single GABAergic cells. The activation of mGluR7 may decrease GABA release to mGluR7-decorated cells at times of high pyramidal cell activity, which elevates extracellular glutamate levels. Alternatively, the presynaptic receptor may be activated by as yet unidentified endogenous ligands released by the GABAergic terminals or the postsynaptic dendrites.

Animals↗

Glycine receptors regulate interneuron differentiation during spinal network development.

Glycinergic and GABAergic excitatory chloride-mediated signaling is often the first form of activity to emerge in the nascent nervous system and has been proposed to be essential for several aspects of nervous system development. However, few studies have examined the effects of disrupting glycinergic transmission. Here we perturbed glycinergic transmission in vivo from the onset of development in zebrafish and examined its impact on the formation of the locomotor circuitry. Targeted knockdown of the embryonic glycine receptor alpha2-subunit disrupted rhythm-generating networks and reduced the frequency of spontaneous glycinergic and glutamatergic events. Immunohistochemistry revealed a reduction in the number of spinal interneurons without affecting sensory and motor neurons. This effect was accompanied by a concomitant increase in the number of mitotic cells, suggesting that glycine receptors regulate interneuron differentiation during early development. Despite the loss of many interneurons, a subthreshold rhythm-generating circuit was still capable of forming. These data provide evidence that glycine receptors, in addition to their role in neurotransmission, regulate interneuron differentiation during development of this central neural network.

Animals↗

Serotonergic control of the hippocampus via local inhibitory interneurons.

Information flow and processing in hippocampal neuronal networks is determined by a wide range of inhibitory mechanisms [e.g., feedforward or feedback, gamma-aminobutyrate (GABA) A or B receptor-mediated, perisomatic shunting, or distal dendritic inhibition], each subserving specialized functions. These forms of local inhibition are mediated by morphologically and neurochemically well-defined, mostly GABA-containing, interneurons, which control large populations of principal cells through their extensive axonal arborizations. These neurons can serve as ideal targets for subcortical pathways, such as those originating in the septum or raphe, which exercise a global control over hippocampal activity. This intriguing possibility prompted us to study whether the profound effect of the serotonergic raphe-hippocampal pathway is mediated by inhibitory interneurons or whether a direct diffuse action on the principal cells is dominant. We demonstrate that axons of this pathway form multiple synaptic contacts with hippocampal GABAergic interneurons. Interestingly, the serotonergic afferents selectively innervate the somata and dendritic trees of GABAergic neurons that contain the 28-kDa calcium-binding protein calbindin D28K, but never those that contain another calcium-binding protein, parvalbumin. These results show that the mechanism by which the serotonergic pathway may exert a powerful influence on hippocampal function involves the modulation of local inhibitory circuits. Furthermore, the selectivity in the choice of target GABAergic interneurons suggests a strong functional specialization among inhibitory circuits, as well as among the subcortical input pathways originating in the septum and raphe.

Afferent Pathways↗

Activity patterns of interneurons in the caudal ganglion of the crayfish.

Responses of ascending interneurons from the caudal ganglion of crayfish have been recorded from single units isolated by dissection from the ventral nerve cord; in addition, post-synaptic activity within the ganglionic neuropile has been studied with intracellular micropipettes. The following classes of interneurons have been found: (1) Large fibers which responded to tactile stimuli with single spikes or phasic bursts. These units usually showed broad receptive fields; and spontaneous activity, when present, showed transitory depressions following responses to natural stimuli. (2) A group of fibers, including many small ones, which responded to proprioceptive stimuli with tonic discharges of varying adaptation rate. (3) Interneurons which showed responses both to tactile stimuli and to activation of the sixth ganglion photoreceptor; and (4) units with constant frequency discharges which were unmodifiable by any of the above afferent inputs. Intracellular recording of post-synaptic activity has shown (1) that widely graded excitatory post-synaptic potentials occur; (2) that multiple firing from single synaptic potentials is usual; (3) that the post-synaptic responses to phasic natural stimuli and to electrical stimulation of ganglionic roots are similar. The existence of widely graded post-synaptic potentials and of extensive receptive fields suggests a high degree of convergence from primary afferents to interneurons. The activation of such post-synaptic units involves integrative synaptic transfer, without 1:1 correspondence between pre- and post-fiber activity.

Animals↗

Sensitivity of sympathetically correlated spinal interneurons, renal sympathetic nerve activity, and arterial pressure to somatic and visceral stimuli after chronic spinal injury.

In the chronic stage of spinal cord injury in humans, both innocuous and noxious somatic and visceral stimuli can elicit severe autonomic dysreflexia characterized by potentially dangerous, sympathetically mediated, increases in arterial pressure. We hypothesized that a similar sympathetic hyperexcitability would be manifested in spinal sympathetic networks of chronically spinally transected rats. To test this hypothesis, we compared the responses of sympathetically correlated spinal interneurons and arterial pressure to both innocuous and noxious stimuli in acutely and chronically spinally transected rats. Experiments were conducted in anesthetized female rats, either within hours of T(3) spinal transection (rats with acute spinal transection) or one month after T(3) spinal transection (rats with chronic spinal transection). Sympathetically correlated spinal interneurons were identified by cross correlating their ongoing activity with simultaneously recorded renal sympathetic nerve activity. Cutaneous stimuli (either light brushing or noxious pinch) were delivered to a wide area of the ipsilateral side of the rat. Colorectal distension was used as a noxious visceral stimulus. The activity of sympathetically correlated interneurons was increased by stimulation of more of the body surface and decreased by stimulation of less of the body surface in rats with chronic spinal transection than in rats with acute spinal transection. Colorectal distension elicited greater increases in arterial pressure in chronically than acutely spinally-transected rats without exciting significantly more interneurons in those rats. These results suggest that spinal circuits undergo significant plastic changes in the chronic stage of spinal cord injury, and they provide a mechanism for the observation, in some human patients, that many stimuli, both noxious and non-noxious, applied caudal to the site of spinal injury increase sympathetic activity and arterial pressure.

Action Potentials↗