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PubMed1976

[Studies on the structure of thalamo-cortical projection neurons and interneurons in the Corpus geniculatum laterale pars dorsalis of albino rats after different histological treatments].

1. In the dorsal part of the lateral geniculate body of the albino rat (Wistar strain) the identification of relay neurons and interneurons has been carried out and the findings in Golgi and Nissl preparations were compared with those in the semithinn sections and in the electron microscopic preparations. 2. The effect of different histolical treatments (perfused by Karnovsky's fluid and embedded in micropal; immersed or perfused by Bouin's fluid, formalin or glutaraldehyde and embedded in celloidine or paraffin) in the structure of the relay neurons and the interneurons in combination with the Nissl method is described. 3. Following immersion the cytoplasm of the relay neurons is relatively broad and has a rough structure in the extreme case (glutaraldehyde) it is vacuolized due to hypoxie or anoxie. In these preparations we could not extimate dark neurons. 4. Following perfusion the cytoplasm of the relay neurons has a fine structure and is more narrow than following fixation by immersion. Furthermore the cell nucleus is enlarged. 5. The identification of interneurons is possible to a high degree of security only after fixation with one of the both aldehydes according to the well-known criteria. When Bouin's fluid is applied the chromatin particle at the nucleolus as one of the important discriminators is not represented. 6. In the single groups the relay neurons were characterized by quantitative parameters. As statistical method of calculation the paired comparison is used. The findings about the semithinn sections are regarded as the standard values because this material is expected to give the most reliable equivalent picture. 7. In 9 out of 12 cases the diameter of the cell soma is smaller and in 3 cases larger than the standard value. 3 values (perfused by Bouin's fluid and embedded in paraffin; immersed by formalin and embedded in celloidine; immersed by glutaraldehyde and embedded in celloidine) do not present remarkable deviation from the standard value. 8. Amongst the diameters of the nuclei only one value is larger than the standard value. 7 values do not significantly differ from the standard value, among them the values of meterial perfused by Bouin's fluid and embedded in paraffin and also immersed by formalin and embedded in celloidine. 9. All the quotientes from soma diameter and nucleus diameter are significantly different to the standard value. The difference between the values of immersion by Bouin's fluid and the standard value is the smallest one. 10. Although the values of soma diameter and nucleus diameter of the mateiral following perfusion by Bouin's fluid and embedding in paraffin as well as following immersion by formalin and embedding incelloidine are not significantly different to the standard value, we prefer immersion by formalin and embedding in celloidine because it is the most suitable method solving the special problems concerning the identification of relay neurons and interneurons in Nissl preparations and the metrical comparisons with findings in Golgi investigations.

Animals↗
PubMed1997

Arachidonic acid inhibits transient potassium currents and broadens action potentials during electrographic seizures in hippocampal pyramidal and inhibitory interneurons.

The transient outward potassium current was studied in outside-out macropatches excised from the soma of CA1 pyramidal neurons and stratum (st.) oriens-alveus inhibitory interneurons in rat hippocampal slices. Arachidonic acid dose dependently decreased the charge transfer associated with the transient current, concomitant with an increase in the rate of current inactivation. Arachidonic acid (AA) did not affect the voltage dependence of steady state inactivation but did prolong the period required for complete recovery from inactivation. The effects of AA were mimicked by the nonmetabolizable analog of AA, 5,8,11,14-eicosatetraynoic acid, suggesting that metabolic products of AA were not responsible for the observed blocking action. In addition, AA blocked st. oriens-alveus-lacunosum-moleculare interneuron transient currents but not currents recorded from basket cell interneurons. In current clamp experiments, AA was without effect on the action potential waveform of CA1 pyramidal neurons under control recording conditions. In voltage-clamp experiments, the use of a test pulse paradigm, designed to mimic the action potential voltage trajectory, revealed that the transient current normally associated with a single spike deactivates too rapidly for AA to have an effect. Transient currents activated by longer duration "action potential" waveforms, however, were attenuated by AA. Consistent with this finding was the observation that AA broadened interictal spikes recorded in the elevated [K+]o model of epilepsy. These data suggest that AA liberated from hippocampal neurons may act to block the transient current selectively in both CA1 pyramidal neurons and inhibitory interneurons and to broaden action potentials selectively under pathological conditions.

5,8,11,14-Eicosatetraynoic Acid↗
PubMed1997

Localization of delta opioid receptor immunoreactivity in interneurons and pyramidal cells in the rat hippocampus.

To study possible cellular targets and subcellular sites of action of opioid ligands in the rat hippocampus, we examined the distribution of the delta opioid receptor (DOR) by immunocytochemistry. By light microscopy, numerous interneurons, or non-principal cells, were intensely labeled for DOR, whereas the CA1 and CA3 pyramidal cells were lightly labeled. DOR-immunoreactive interneurons were found throughout the hippocampus but were particularly concentrated in stratum oriens of the CA1 region. Double labeling immunofluorescence revealed that DOR-immunoreactivity was found in a subpopulation of gamma-aminobutyric acid (GABA)-containing interneurons, which included most somatostatin-immunoreactive cells. Electron microscopic analysis of sections singly labeled for DOR revealed that DOR-immunoreactive profiles were abundant and widespread throughout all hippocampal lamina and had a similar distribution in CA1 and CA3. DOR-immunoreactivity was sometimes found in dendrites, which corresponded in morphology to those of interneurons. In addition, DOR-labeling was found in the shafts and spines of many dendrites, which exhibited the morphology of pyramidal cell dendrites. Within dendrites, dense DOR-immunoreactivity was associated with the plasmalemmal surface at or near the postsynaptic density, usually of asymmetric synapses. In addition, DOR labeling was present in a heterogeneous population of axon terminals, as well as in astrocytic profiles. At mossy fiber synapses, DOR labeling was occasionally found at both pre-and post-synaptic sites. These studies demonstrate that DOR is present at multiple sites on diverse cell types where it may function to regulate neuronal activity in the hippocampus.

Animals↗
PubMed1997

[Interneuronal interrelationships in autonomic ganglia (the ultrastructural aspects)].

Ultrastructural and immunocytochemical results of studies on interneuronal relationship in vegetative ganglia were represented. Age-related changes of synapses (appearance of synaptic type vesicles and smooth endoplasmic reticulum cisterns in postsynaptic terminals), peculiarities of interneuronal links in neurons containing neuropeptide Y and NO-synthase and structural manifestations of non-synaptic interneuronal links were shown. Modern views on interneuronal communications were discussed.

Aging↗
PubMed1997

Cholinergic and GABAergic interneurons in the striatum.

In the striatum, interneurons have not been as well characterized physiologically as the spiny projection cells. We found that the neostriatal interneurons can be divided at least into three classes by physiological, chemical and morphological criteria. The first was FS cells (fast-spiking cells) which fired very short-duration action potentials at constant spike frequency during depolarizing pulses, were immunoreactive for parvalbumin (calcium-binding protein), and had axons with very dense collateralization within or near their dendritic fields. Another class was identified as those which fired low-threshold spikes (LTS cells) from hyperpolarized potentials, were positive for somatostatin and nitric oxide synthase (NOS), and had the largest axonal fields. The other class of interneurons had longer-lasting afterhyperpolarizations (LA cells), were positive for choline acetyltransferase, and were mostly large aspiny cells. Glutamic acid decarboxylase (GAD67) or GABA immunoreactivity was detected at the somata or terminals of parvalbumin FS cells and somatostatin/NOS LTS cells, but not of cholinergic LA cells. Substance P, probably released from the collaterals of cells projecting to the substantia nigra, excited LA cells and LTS cells, but not FS cells. These results suggest that the striatum has at least one type of cholinergic and two types of GABAergic interneurons which are different in physiological, chemical and pharmacological characteristics.

Acetylcholine↗
PubMed1997

Functions for interneuronal nets in the hippocampus.

Recent advances in the physiology of hippocampal interneurons are summarized in this article. These findings suggest that through their interconnectivity inhibitory interneurons can maintain large-scale oscillations at various frequency ranges (theta, gamma, and 200-Hz bands). We suggest that networks of inhibitory interneurons within the forebrain impose coordinated oscillatory "contexts" for the "content" carried by networks of principal cells. These oscillating inhibitory networks may provide the precise temporal structure necessary for ensembles of neurons to perform specific functions, such as memory trace formation and retrieval. In addition, synaptic inhibition is shown to reduce the somadendritic backpropagation of sodium spikes and to prevent the occurrence of calcium spikes in dendrites. These observations indicate that interneurons are in an excellent position to control neuronal plasticity and allow synaptic transmission either with or without long-term modification of synaptic strength.

Hippocampus↗
PubMed1997

Do GABAA and GABAB inhibitory postsynaptic responses originate from distinct interneurons in the hippocampus?

GABAergic inhibition of hippocampal pyramidal cells is mediated by two distinct subtypes of postsynaptic receptors, GABAA and GABAB. Electrical stimulation of inhibitory cells or fibres in the CA1 subfield of the hippocampus yields a biphasic inhibitory postsynaptic potential (IPSP) in pyramidal cells, consisting of an early GABAA- and a late GABAB-mediated component. CA1 interneurons are a heterogeneous population of cells, which differ on the basis of their morphology, physiological properties, target selectivity onto principal cells, and network connectivity. Inhibitory synaptic circuitry appears to be specialized, since feedback inhibition may invoke only postsynaptic GABAA receptors, whereas feedforward inhibition may invoke both postsynaptic GABAA and GABAB receptors. In this review, we examine the evidence for and against the notion that distinct interneurons may be responsible for GABAA- and GABAB-mediated inhibition. Overall, the evidence suggests that (i) certain interneurons may generate solely GABAA inhibition, but the available data do not distinguish whether other interneurons mediate (ii) solely GABAB inhibition or (iii) a combination of both GABAA and GABAB.

Feedback↗
PubMed1997

Expression patterns of developmental control genes in normal and Engrailed-1 mutant mouse spinal cord reveal early diversity in developing interneurons.

The vertebrate spinal cord has long served as a useful system for studying the pattern of cell differentiation along the dorsoventral (d/v) axis. In this paper, we have defined the expression of several classes of genes expressed in restricted d/v domains in the intermediate region (IR) of the mouse spinal cord, in which most interneurons are generated. From this analysis, we have found that spinal cord interneurons and their precursors express unique combinations of transcription factors and Notch ligands at the onset of their differentiation. The domains of expression of a number of different classes of genes share similar boundaries, indicating that there could be a basic subdivision of the ventral IR into four distinct regions. This differential gene expression suggests that spinal cord interneurons acquire unique identities early in their development and that Notch signaling mechanisms may participate in the determination of cell fate along the d/v axis. Gene expression studies in Engrailed-1 (En-1) mutants showed that En-1-expressing and other closely positioned classes of neurons do not require the homeodomain protein En-1 for their early pattern of differentiation. Rather, it is suggested that En-1 may function to distinguish a subset of interneurons during the later maturation of the spinal cord.

Animals↗
PubMed1998

Opioid inhibition of hippocampal interneurons via modulation of potassium and hyperpolarization-activated cation (Ih) currents.

The actions of mu- and delta-opioid agonists (DAMGO and DPDPE, respectively) on GABAergic interneurons in stratum oriens of area CA1 of the hippocampus were examined by using whole-cell voltage-clamp recordings in brain slices. Both agonists consistently generated outward currents of similar magnitude (15-20 pA) in the majority of cells. However, under control conditions, current-voltage (I/V) relationships revealed that only a small number of these cells (3 of 77) demonstrated clear increases in membrane conductance, associated with the activation of the potassium current known as Girk. These interneurons also exhibited a slowly activating, inwardly rectifying current known as Ih on hyperpolarizing step commands. Ih was blocked by the extracellular application of cesium (3-9 mM) or ZD 7288 (10-100 microM) but was insensitive to barium (1-2 mM). In an effort to determine whether the holding current changes were attributable to the modulation of Girk and/or Ih, we used known blockers of these ion channels (barium or cesium and ZD 7288, respectively). Extracellular application of cesium (3-9 mM) or ZD 7288 (25-100 microM) blocked Ih and significantly reduced the opioid-induced outward currents by 58%. Under these conditions the opioid agonists activated a potassium current with characteristics similar to Girk. Similarly, during barium (1-2 mM) application the opioid-induced outward currents were reduced by 46%, and a clear reduction in Ih and the whole-cell conductance was revealed. These data suggest that the opioids can modulate both Ih and Girk in the same population of stratum oriens interneurons and that the modulation of these ion channels can contribute to the inhibition of interneuron activity in the hippocampus.

Analgesics↗
PubMed1998

Selective innervation of neostriatal interneurons by a subclass of neuron in the globus pallidus of the rat.

A subpopulation of neurons in the globus pallidus projects to the neostriatum, which is the major recipient of afferent information to the basal ganglia. Given the moderate nature of this projection, we hypothesized that the pallidostriatal projection might exert indirect but powerful control over principal neuron activity by targeting interneurons, which comprise only a small percentage of neostriatal neurons. This was tested by the juxtacellular labeling and recording of pallidal neurons in combination with immunolabeling of postsynaptic neurons. In addition to innervating the subthalamic nucleus and output nuclei, 6 of 23 labeled pallidal neurons projected to the neostriatum. Both the firing characteristics and the extent of the axonal arborization in the neostriatum were variable. However, light and electron microscopic analysis of five pallidostriatal neurons revealed that each neuron selectively innervated neostriatal interneurons. A large proportion of the boutons of an individual axon (19-66%) made contact with parvalbumin-immunoreactive interneurons. An individual parvalbumin-immunoreactive neuron (n = 27) was apposed on average by 6.7 boutons (SD = 6.1) from a single pallidal axon (n = 2). Individual pallidostriatal boutons typically possessed more than one symmetrical synaptic specialization. In addition, 3-32% of boutons of axons from four of five pallidal neurons contacted nitric oxide synthase-immunoreactive neurons. Descending collaterals of pallidostriatal neurons were also found to make synaptic contact with dopaminergic and GABAergic neurons of the substantia nigra. These data imply that during periods of cortical activation, individual pallidal neurons may influence the activity of GABAergic interneurons of the neostriatum (which are involved in feed-forward inhibition and synchronization of principle neuron activity) while simultaneously patterning neuronal activity in basal ganglia downstream of the neostriatum.

Action Potentials↗
PubMed2001

Substance P and nitric oxide signaling in cerebral cortex: anatomical evidence for reciprocal signaling between two classes of interneurons.

Parvalbumin-containing fast-spiking interneurons in the cerebral cortex exhibit widespread electrical coupling, as do somatostatin-containing low-threshold spiking interneurons. Besides the classical neurotransmitter gamma-aminobutyric acid, these cortical interneurons may also release various neuropeptides including substance P (SP), as well as the freely diffusible messenger nitric oxide (NO). To investigate whether these two networks of interneurons might interact via these nonclassical messengers, we performed immunocytochemistry for SP and NO signaling pathways in rat somatic sensory cortex. SP was found in a subset of parvalbumin-positive cells concentrated in layers IV and V, whereas its receptor, NK1, was found in a subset of somatostatin-containing neurons (and also, at much lower levels, in a disjoint subset of parvalbumin-containing neurons). Only 4% of SP-containing axon terminals were apposed to NK1-positive dendrites, suggesting that in the cerebral cortex, SP may act predominantly as a paracrine neuromediator. Nitric oxide synthase-I (NOS-I), the synthetic enzyme for NO, was found almost exclusively in NK1-positive neurons; 95% of intensely somatostatin/NK1-positive neurons were also positive for NOS-I, and 94% of NOS-positive neurons were also positive for NK1. Immunoreactivity for soluble guanylyl cyclase (the NO receptor) was at high levels in the apical dendrites of layer V pyramidal neurons and in parvalbumin/SP-positive neurons. These data point to a novel reciprocal chemical interaction between two inhibitory networks in the rat neocortex.

Animals↗
PubMed1989

Synaptic connections between corticogeniculate axons and interneurons in the dorsal lateral geniculate nucleus of the cat.

Anatomical evidence is provided for direct synaptic connections by axons from visual cortex with interneurons in lamina A of the cat's dorsal lateral geniculate nucleus. Corticogeniculate axon terminals were labeled selectively with 3H-proline and identified by means of electron microscopic autoradiography. Interneurons in the lateral geniculate nucleus were stained with antibodies that had been raised against gamma aminobutyric acid (GABA). We found that corticogeniculate terminals synapsed with dendrites stained positively for GABA about three times as often as with unstained dendrites. Of the corticogeniculate terminals that contacted GABA-positive dendrites, 97% made synaptic connections with dendritic shafts. Only 3% synapsed with F profiles, the vesicle-filled dendritic appendages characteristic of lateral geniculate interneurons. These results suggest that the corticogeniculate pathway in the cat is directed primarily at interneurons and is organized synaptically to influence the integrated output of these cells, rather than the local interactions in which their dendritic specializations participate.

Animals↗
PubMed2000

Directional sensitivity of an identified wind-sensitive interneuron during the postembryonic development of the locust.

Simultaneous extracellular recordings from both locust abdominal connectives show a differential activation of both bilateral homologues of an identified long projection interneuron (A4I1) in response to wind stimuli from different directions. Despite the previously shown extensive structural dynamics of sensory afferents and synaptic rearrangement of the direct afferent-to-interneuron connections during postembryonic development, a directional sensitivity is already present in first instar nymphs. Only quantitative changes in the strength of the directional response can be detected. Intracellular stainings of the A4I1 interneuron in first instar nymphs and adults show that general morphological features do not change during postembryonic development, in contrast to the presynaptic sensory afferents. This also holds for general morphological features of pleuroaxillary flight motoneurons. The output connections of A4I1 to these motoneurons and an unidentified intersegmental interneuron are already present in flightless nymphs.

Journal Article↗
PubMed2002

Fast synaptic inhibition promotes synchronized gamma oscillations in hippocampal interneuron networks.

Networks of GABAergic interneurons are of critical importance for the generation of gamma frequency oscillations in the brain. To examine the underlying synaptic mechanisms, we made paired recordings from "basket cells" (BCs) in different subfields of hippocampal slices, using transgenic mice that express enhanced green fluorescent protein (EGFP) under the control of the parvalbumin promoter. Unitary inhibitory postsynaptic currents (IPSCs) showed large amplitude and fast time course with mean amplitude-weighted decay time constants of 2.5, 1.2, and 1.8 ms in the dentate gyrus, and the cornu ammonis area 3 (CA3) and 1 (CA1), respectively (33-34 degrees C). The decay of unitary IPSCs at BC-BC synapses was significantly faster than that at BC-principal cell synapses, indicating target cell-specific differences in IPSC kinetics. In addition, electrical coupling was found in a subset of BC-BC pairs. To examine whether an interneuron network with fast inhibitory synapses can act as a gamma frequency oscillator, we developed an interneuron network model based on experimentally determined properties. In comparison to previous interneuron network models, our model was able to generate oscillatory activity with higher coherence over a broad range of frequencies (20-110 Hz). In this model, high coherence and flexibility in frequency control emerge from the combination of synaptic properties, network structure, and electrical coupling.

Animals↗
PubMed1992

GABAergic Interneurons are the Major Postsynaptic Targets of Median Raphe Afferents in the Rat Dentate Gyrus.

The termination pattern of median raphe axons was studied in the rat dentate gyrus using Phaseolus vulgaris leucoagglutinin as an anterograde tracer, in combination with postembedding immunostaining for gamma-amino-butyric acid (GABA), and pre-embedding immunostaining for calbindin D28k, parvalbumin and GABA. Postembedding immunogold staining for GABA revealed that the majority (73.7%) of anterogradely labelled median raphe boutons make synaptic contacts with GABA-immunoreactive postsynaptic targets, mainly with dendritic shafts and perikarya. Pre-embedding immunocytochemical double staining for the anterograde tracer and GABA confirmed the electron microscopic results and showed that varicose median raphe axons establish multiple contacts with fusiform interneurons in the hilus and different types of basket cells in the granule cell layer. Some of the innervated cells were shown to contain calbindin D28k, whereas GABAergic interneurons containing another calcium-binding protein, parvalbumin, were never seen to receive multiple contacts from axons of raphe origin. Our results suggest that serotonergic median raphe fibres influence the firing of dentate granule cells via local inhibitory interneurons. The mechanism of using these interneurons with extensive local connections as monosynaptic targets may explain the great efficacy of this pathway in the control of hippocampal electrical activity.

Journal Article↗
PubMed1997

Evidence that spinal interneurons undergo programmed cell death postnatally in the rat.

Programmed cell death has been demonstrated in several specific neuronal populations as a mechanism for modulating the population size following differentiation, but its applicability to all neuronal types is unclear. Evidence for programmed cell death in some populations such as the numerous spinal interneurons has been lacking. We have studied the incidence of apoptosis in the rat spinal cord with three different methods and found a previously undocumented wave of apoptosis occurring in spinal grey matter shortly after birth. The apoptotic morphology was confirmed ultrastructurally. Dying cells were identified as neurons by immunocytochemical labelling for neuronal markers and had an anatomical distribution which indicated that most of the apoptotic cells were interneurons not motoneurons. This wave of apoptosis has the characteristics of a discrete developmental process and occurs later than that of either ventral horn motoneurons or dorsal root ganglion cells, to which most spinal interneurons are connected. These findings indicate that interneurons do undergo programmed cell death, and we suggest that this occurs in response to the earlier reduction in size of their main synaptic targets.

Aging↗
PubMed1994

ACTIVITY OF GIANT INTERNEURONES AND OTHER WIND-SENSITIVE ELEMENTS OF THE TERMINAL GANGLION IN THE WALKING CRICKET

Using intracellular recording techniques in stationary walking crickets (Gryllus bimaculatus), we have investigated the relationship between locomotion and the activity of interneurones ascending from the terminal ganglion. Nine different types of giant interneurones (GI) were characterized during walking and standing. One third of them reduced their activity, while the others enhanced their spike rate, during walking. These physiological properties were strictly correlated with morphological characteristics such as axon position in the longitudinal tracts of the terminal ganglion. In general, ventral GIs reduced and dorsal GIs increased their spike frequency during walking. In some of them, there was a weak but significant correlation between the spike rate and translational speed, but no correlation with rotational speed. In all GIs except 10-3a, the changes in activity occurred at the start of walking. In GI 10-3a, an increase in membrane potential and spike rate was observed before the start of locomotion. Therefore, an intrinsic mechanism within the central nervous system operating on GI 10-3a is suggested. Additionally, the activities of filiform hair receptors and of previously undescribed small ascending interneurones (SAI) have been studied during walking. About 80 % of the receptors slightly increased their spike rate during walking, while one SAI became more active during walking and another one was hardly affected. The physiological properties of ascending interneurones are discussed with respect to their modulation and particular function during walking.

Journal Article↗
PubMed2000

Development, neurochemical properties, and axonal projections of a population of last-order premotor interneurons in the white matter of the chick lumbosacral spinal cord.

There is general agreement that last-order premotor interneurons-a set of neurons that integrate activities generated by the spinal motor apparatus, sensory information and volleys arising from higher motor centres, and transmit the integrated signals to motoneurons through monosynaptic contacts-play crucial roles in the initiation and maintenance of spinal motor activities. Here, we demonstrate the development, neurochemical properties, and axonal projections of a unique group of last-order premotor interneurons within the ventrolateral aspect of the lateral funiculus of the chick lumbosacral spinal cord. Neurons expressing immunoreactivity for neuron-specific enolase were first detected in the ventrolateral white matter at embryonic day 9 (E9). The numbers of immunoreactive neurons were significantly increased at E10-E12, while most of them were gradually concentrated in small segmentally arranged nuclei (referred to as major nuclei of Hofmann) protruding from the white matter in a necklace like fashion dorsal to the ventral roots. The major nuclei of Hofmann became more prominent at E12-E16, but substantial numbers of cells were still located within the ventrolateral white matter (referred to as minor nucleus of Hofmann). The distribution of immunoreactive neurons achieved by E16 was maintained during later developmental stages and was also characteristic of adult animals. After injection of Phaseolus vulgaris-leucoagglutinin unilaterally into the minor nucleus of Hofmann, labeled fibres were detected in the ventrolateral white matter ipsilateral to the injection site. Ascending and descending fibres were revealed throughout the entire rostro-caudal length of the lumbosacral spinal cord. Axon terminals were predominantly found within the lateral motor column and the ventral regions of lamina VII ipsilateral to the injection site. Several axon varicosities made close appositions with somata and dendrites of motoneurons, which were identified as synaptic contacts in a consecutive electron microscopic study. With the postembedding immunogold method, 21 of 97 labeled terminals investigated were immunoreactive for glycine and 2 of them showed immunoreactivity for gamma-aminobutyric acid (GABA). The axon trajectories of neurons within the minor nucleus of Hofmann suggest that some of these cells might represent a population of last-order premotor interneurons. J. Exp. Zool. 286:157-172, 2000.

Animals↗