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Anatomy of the ocellar interneurons of acridid grasshoppers. I. The large interneurons.

The anatomy of the large ocellar interneurons in the brain of five species of acridid grasshoppers of two different subfamilies (Schistocerca vaga, S. gregaria, Gastrimargus africanus, Trimerotropis pallidipennis, and Arphia conspersa) was revealed by cobalt-filling of the three ocellar nerves and subsequent reconstructions from silver-intensified (Timm's method) serial sections. Conflicts in the literature are reviewed (Tables 1, 2) and differences in the number of cells, anatomical descriptions of these cells, and nomenclature are resolved by demonstration of an identical number of large ocellar identical number of large ocellar interneurons in all five species examined (Fig. 1). There are 17 large 1st-order ocellar interneurons (Figs. 2, 3). Each of the three ocellar nerves contains the axons of seven large interneurons; four of these interneurons have axons in two ocellar nerves. The anatomy of three pairs of 2nd-order ocellar interneurons (with branches in the ocellar tracts within the brain and axons in the circum-esophageal connectives) is reconsidered in light of recent conflicts in the literature. Previous accounts by Williams (1975) of interneurons O2, O3, and PI(2):5 are corroborated and new details added (Fig. 7) by the use of a cobalt method that appears to stain these 2nd-order interneurons transsynaptically (Fig. 6).

Animals↗

Spinal interneurones; how can studies in animals contribute to the understanding of spinal interneuronal systems in man?

The first part of this review deals with arguments that the essential properties and organization of spinal interneuronal systems in the cat and in man are similar. The second part is concerned with the possibility that some interneuronal systems may be responsible for motor disturbances caused by spinal cord injuries and that these interneurones may be defined. This possibility is discussed with respect to the hyperexcitability of alpha-motoneurones and the exaggeration of stretch reflexes in spastic patients. To this end, what is known about cat spinal interneurones and about the neuronal basis and pharmacological treatment of spasticity, is put together. Interneurones in di- and trisynaptic reflex pathways from group II muscle afferents are singled out, since they are depressed by the alpha(2) noradrenaline receptor agonists clonidine and tizanidine, which is a critical feature of interneurones expected to contribute to exaggerated stretch reflexes which are reduced by alpha(2) noradrenaline receptor agonists. Recent observations that reflex excitation of extensor motoneurones from group II afferents is enhanced in spastic patients and that the pathologically strong reflex actions of group II afferents are reduced by clonidine and tizanidine support this proposal. On the other hand, a lack of effect of clonidine and tizanidine upon other types of excitatory or inhibitory interneurones argues against any major contribution of such interneurones to the abnormally strong responses of alpha-motoneurones to muscle stretch.

Animals↗

Different populations of vasoactive intestinal polypeptide-immunoreactive interneurons are specialized to control pyramidal cells or interneurons in the hippocampus.

The postsynaptic targets of three vasoactive intestinal polypeptide-containing GABAergic interneuron types were examined in the rat hippocampus. Two of them showed remarkable target selectivity for other GABAergic neurons, while the third contacted the somata and proximal dendrites of pyramidal cells. Vasoactive intestinal polypeptide-positive interneurons innervating the stratum oriens/alveus border in the CA1 region were shown to establish multiple contacts with horizontal GABAergic interneurons immunoreactive for type 1 metabotropic glutamate receptor. Similarly, identified axons of vasoactive intestinal polypeptide-positive interneurons projecting to stratum radiatum were found to establish symmetrical synapses largely on GABAergic dendrites. The majority of these postsynaptic GABAergic neurons were shown to contain calbindin or vasoactive intestinal polypeptide. In contrast to the first two vasoactive intestinal polypeptide-containing cell populations, vasoactive intestinal polypeptide-positive interneurons arborizing in stratum pyramidale formed baskets around pyramidal cells. These results revealed a new element in cortical microcircuits, interneurons which are specialized to innervate other GABAergic interneurons. The role of this new component may be the synchronization of dendritic inhibition, or an input-specific disinhibition of pyramidal cells in various dendritic domains. In contrast, vasoactive intestinal polypeptide-containing basket cells are likely to be involved in perisomatic inhibition of pyramidal neurons, and represents a new basket cell type different from that containing parvalbumin.

Animals↗

Spiking local interneurones in the mesothoracic ganglion of the locust: homologies with metathoracic interneurones.

Two bilaterally symmetrical groups of spiking local interneurones have been characterized in the mesothoracic ganglion of the locust. The cell bodies of one group, the "midline group," lie at the ventral midline. Their primary neurites run in the ventral loop of ventral commissure II to form extensive branches in the neuropile of one-half of the ganglion. A dorso-ventral process in the perpendicular tract links two distinct fields of branches, one ventral and consisting of numerous fine branches of a uniform texture that arise from stout secondary neurites, and the other more dorsal consisting of fewer branches of a varicose appearance. Cell bodies of the second, the "anterior-lateral" group, lie close to the lateral edge of an anterior connective. Their primary neurites run in a more anterior ventral commissure and their neuropilar branches are divided into two fields by a process in a more anterior dorso-ventral tract. Within the two groups, each interneurone has its own distinctive shape that is an elaboration on these basic plans. Each interneurone also has its own characteristic physiology, being excited by a particular array of mechanoreceptors on the middle leg on the same side of the body as its neuropilar branches. The receptive fields of the interneurones, defined in this way, can be extensive and cover a particular surface of all parts of the leg, or restricted to one surface of, for example, the tarsus. These interneurones therefore bear a striking resemblance to two groups of spiking local interneurones in the adjacent segmental ganglion of the metathorax.

Action Potentials↗

Wind-activated thoracic interneurons of the cockroach: II. Patterns of connection from ventral giant interneurons.

A number of thoracic interneurons (TIs) have been found to receive inputs from ventral giant interneurons (vGIs). Each of these cells responds to wind with short latency depolarizations. The previous paper described response properties of several TIs to wind stimuli, including those excited by vGIs. The data showed a correlation between the shape of the TI's wind fields and its morphology. The presence of ventral branches located near the midline of the ganglion predicts a strong response to wind on that side. These ventral median (VM) branches are in the proper location to permit overlap with processes from vGIs. Here we describe the patterns of connections between individual vGIs and 13 of the thoracic interneurons located in the meso- and metathoracic ganglia. A correlation was found between the presence of VM branches and excitation by vGIs. TIs were only excited by vGIs on the side(s) on which VM branches exist. However, presence of a VM branch does not imply that all vGIs on that side make connections with the TI. Summation was found between various vGIs that excited each individual thoracic interneuron. In unilateral thoracic interneurons, no sign of inhibition was found from vGIs on the sides opposite that which contained excitatory vGI axons. Neither was there any evidence of inhibition from dorsal giant interneurons. In addition preliminary evidence indicated that left-right homologues do not inhibit one another. Thus, the data suggest that directional wind fields are primarily the result of selective connection from specific vGIs.

Action Potentials↗

Localization of ventral giant interneuron connections to the ventral median branch of thoracic interneurons in the cockroach.

A detailed morphological study was performed to localize the probable sites of connections between two identified populations of interneurons (ventral giant interneurons and type-A thoracic interneurons) in the cockroach. Type-A thoracic interneurons (TIAS) appear to play an important role in orienting the cockroach during wind-mediated escape. However, their large number, approximately 100 neurons, precludes analyzing each cell's role electrophysiologically. The TIAS are characterized by a prominent branch located on one or both sides of the ventral median (VM) region of the thoracic ganglion in which their soma resides. The presence of this ventral median branch can be used to predict connectivity with left or right ventral giant interneurons (vGIs) (Ritzmann and Pollack, 1988) and is correlated with the TIA's directional response to wind (Westin, Ritzmann, and Goddard, 1988), suggesting that this is the locus of synaptic connection. Two approaches were employed to address this hypothesis. Morphological overlap of differentially labelled cells (ethidium bromide, Lucifer Yellow) was examined at the light microscopic level to locate areas of possible synaptic contact. Experiments were also performed in which one-half of the vGI input to the TIAs was surgically removed early in postembryonic development. Although no changes in the overall branching pattern were observed, the VM branches on the operated side were significantly shorter than were those on the unoperated side. Thoracic interneurons that do not receive inputs from vGIs were unaffected by this surgery. The data reported here thereby confirm previous observations by localizing the vGI inputs specifically to the VM branch, and provide a morphological cue for predicting connectivity and function.

Animals↗

Enkephalin-containing interneurons are specialized to innervate other interneurons in the hippocampal CA1 region of the rat and guinea-pig.

Enkephalins are known to have a profound effect on hippocampal inhibition, but the possible endogenous source of these neuropeptides, and their relationship to inhibitory interneurons is still to be identified. In the present study we analysed the morphological characteristics of met-enkephalin-immunoreactive cells in the CA1 region of the rat and guinea-pig hippocampus, their coexistence with other neuronal markers and their target selectivity at the light and electron microscopic levels. Several interneurons in all subfields of the hippocampus were found to be immunoreactive for met-enkephalin. In the guinea-pig, fibres arising from immunoreactive interneurons were seen to form a plexus in the stratum oriens/alveus border zone, and basket-like arrays of boutons on both enkephalin-immunoreactive and immunonegative cell bodies in all strata. Immunoreactive boutons always established symmetric synaptic contacts on somata and dendritic shafts. Enkephalin-immunoreactive cells co-localized GABA, vasoactive intestinal polypeptide and calretinin. Postembedding immunogold staining for GABA showed that all the analysed enkephalin-immunoreactive boutons contacted GABAergic postsynaptic structures. In double-immunostained sections, enkephalin-positive axons were seen to innervate calbindin D28k-, somatostatin-, calretinin- and vasoactive intestinal polypeptideimmunoreactive cells with multiple contacts. Based on these characteristics, enkephalin-containing cells in the hippocampus are classified as interneurons specialized to innervate other interneurons, and represent a subset of vasoactive intestinal polypeptide- and calretinin-containing cells. The striking match of ligand and receptor distribution in the case of enkephalin-mediated interneuronal communication suggests that this neuropeptide may play an important role in the synchronization and timing of inhibition involved in rhythmic network activities of the hippocampus.

Animals↗

Hippocampal pyramidal cell-interneuron spike transmission is frequency dependent and responsible for place modulation of interneuron discharge.

The interplay between principal cells and interneurons plays an important role in timing the activity of individual cells. We investigated the influence of single hippocampal CA1 pyramidal cells on putative interneurons. The activity of CA1 pyramidal cells was controlled intracellularly by current injection, and the activity of neighboring interneurons was recorded extracellularly in the urethane-anesthetized rat. Spike transmission probability between monosynaptically connected pyramidal cell-interneuron pairs was frequency dependent and highest between 5 and 25 Hz. In the awake animal, interneurons were found that had place-modulated firing rates, with place maps similar to their presynaptic pyramidal neuron. Thus, single pyramidal neurons can effectively determine the firing patterns of their interneuron targets.

Action Potentials↗

Interneuron Diversity series: Molecular and genetic tools to study GABAergic interneuron diversity and function.

Structural and functional diversity of GABAergic interneurons has become increasingly central in our understanding of the elemental steps of information processing in the brain. The use of different molecular, electrophysiological and anatomical techniques has provided a wealth of new information regarding GABAergic interneurons over the past decade but it has also led to confusion regarding the number of subtypes of GABAergic interneurons. Combinatorial approaches that also consider multiple parameters seem now to offer renewed hope for finally clarifying the structural diversity of GABAergic interneurons. New molecular techniques have become a powerful tool for exposing the functional diversity of GABAergic neurons at the cellular, microcircuit and systems levels. This article reviews literature regarding molecular tools that have been used, or that appear promising for future attempts, to classify GABAergic interneurons. Some important limitations will also be indicated.

Animals↗

Cholinergic septal afferent terminals preferentially contact neuropeptide Y-containing interneurons compared to parvalbumin-containing interneurons in the rat dentate gyrus.

Septal cholinergic neurons may affect hippocampal memory encoding and retrieval by differentially targeting parvalbumin (PARV)-containing basket cells and neuropeptide Y (NPY) interneurons. Thus, the cellular associations of cholinergic efferents, identified by the low-affinity, p75 neurotrophin receptor (p75(NTR)), with interneurons containing either PARV or NPY in the hilus of the rat dentate gyrus were examined in single sections using dual labeling immunoelectron microscopy. Most profiles immunoreactive (IR) for PARV and NPY were perikaryal and dendritic and found within the infragranular and central hilar regions, respectively, whereas most profiles with p75(NTR)-labeling were unmyelinated axons and axon terminals. Although PARV-labeled profiles were more numerous, p75(NTR)-labeled axons and terminals contacted few PARV-IR profiles compared to NPY-labeled profiles (2% of 561 for PARV vs 12% of 433 for NPY). Moreover, structures targeted by p75(NTR)-IR axon terminals varied depending on the presence of PARV or NPY immunoreactivity. p75(NTR)-IR terminals primarily contacted PARV-IR dendrites (87%) compared to somata (13%); however, they contacted more NPY-IR somata (57%) than dendrites (43%). p75(NTR)-labeled terminals formed exclusively symmetric (inhibitory-type) synapses with PARV-IR somata and dendrites; however, they formed mostly symmetric but also asymmetric (excitatory-type) synapses with NPY-IR somata and dendrites. These results suggest that septal cholinergic efferents in the dentate gyrus: (1) preferentially innervate NPY-containing interneurons compared to PARV-containing basket cells; and (2) may provide a more powerful (i.e., somatic contacts), yet functionally diverse (i.e., asymmetric and symmetric synapses), modulation of NPY-containing interneurons. Moreover, they provide evidence that neurochemical subsets of hippocampal interneurons can be distinguished by afferent input.

Afferent Pathways↗

Synaptic connections of dorsal horn group II spinal interneurons: synapses formed with the interneurons and by their axon collaterals.

Five dorsal horn interneurons with monosynaptic input from group II primary afferent fibres were physiologically characterized and intracellularly labelled with horseradish peroxidase. The cells were prepared for combined light and electron microscopy, and synaptic arrangements formed by axon collaterals of interneurons and synapses formed with their dendrites and somata were examined with the electron microscope. Immunogold reactions for gamma-aminobutyric acid, glycine and glutamate were performed to determine if these synapses were excitatory or inhibitory. Axon collaterals in lamina VI formed synapses with somata and dendrites of other neurons, and collaterals of one cell also formed axoaxonic synapses. It was concluded that one cell from the sample was inhibitory, whereas the remainder were probably excitatory. Dendrites and cell bodies of interneurons were contacted by several types of synaptic bouton. The first type of bouton displayed immunoreactivity for glutamate, the second type contained both gamma-aminobutyric acid and glycine, the third type contained glycine alone, and the fourth type contained gamma-aminobutyric acid alone. Some large glutamatergic boutons were postsynaptic to other boutons. Presynaptic boutons at these axoaxonic synapses always contained gamma-aminobutyric acid but a minority also contained glycine. The results of this study demonstrate the heterogeneity of dorsal horn group II interneurons and provide evidence that they include inhibitory and probably also excitatory neurons. Boutons originating from several chemically different classes of neuron are responsible for postsynaptic inhibition of these interneurons, and the presence of axoaxonic synapses indicates that their excitatory input is also controlled presynaptically.

Animals↗

Local commissural interneurons integrate information from intersegmental coordinating interneurons.

The information that coordinates movements of swimmerets on different segments of the crayfish abdomen is conducted by interneurons that originate in each abdominal ganglion. These interneurons project axons to neighboring ganglia and beyond. To discover the anatomy of these axons in their target ganglia, we used Neurobiotin and dextran-Texas Red microelectrodes to fill them near their targets. Coordinating axons coursed through these target ganglia close to the midline and extended only a few short branches that did not approach the lateral neuropils. Two of the three types of coordinating axons made direct synaptic connections with a class of local commissural interneurons that relayed the information to targets in the swimmeret pattern-generating circuits. These commissural interneurons, named here ComInt 1 neurons, followed a particular route to cross the midline and reach their targets. ComInt 1 neurons were nonspiking; they received EPSPs from the coordinating axons near the midline and transmitted this information to their targets in the lateral neuropils using graded transmission. The output of each ComInt 1 was restricted to a single local circuit and had opposite effects on the power-stroke and return-stroke motor neurons driven by that circuit. ComInt 1 neurons were direct postsynaptic targets of both descending and ascending coordinating axons that originated in other anterior and posterior ganglia. Because of phase differences in the impulses in these different coordinating axons, their signals arrived simultaneously at each ComInt 1. We discuss these findings in the context of alternative models of the intersegmental coordinating circuit.

Animals↗

Anatomy of the ocellar interneurons of acridid grasshoppers. II. The small interneurons.

The anatomy of the small ocellar interneurons in the brain of the acridid grasshopper Schistocerca vaga was revealed by cobalt-filling the three ocellar nerves and subsequent reconstructions from silver-intensified (Timm's method) serial sections. In total, 61 small ocellar interneurons were repeatedly identified with arborizations in many areas of the brain and optic lobe, including in particular the posterior neuropil, ocellar tracts, protocerebral bridge, lobula, ventral bridge and tritocerebral crotch, calyces, and antenno-glomerular tracts. Each ocellar nerve contains the axons of small cells that arborize in the other two ocellar tracts; these tracts are sites of ocellar integration. Direct interactions between the ocelli and compound eyes are suggested by the projections of small ocellar interneurons into the proximal lobula. Small cell arborizations from all three ocelli are distributed actoss much of the protocerebral bridge, implying a role for the bridge as an ocellar neuropil within the brain. Four of the small interneurons could be seen in whole-mount preparations and are demonstrated to be identical in five species of acridid grasshoppers of two different subfamilies: Schistocera vaga, S. gregaria, Gastrimargus africanus, Trimerotropis pallidipennis, and Arphia conspersa.

Animals↗

Inhibition of crossed caudal interneurons by lateral interneurons in lamprey spinal chord during fictive locomotion.

Templates of the membrane potential profiles from lateral (LI) interneurons and motoneurons during glutamate- and N-methyl-D-aspartate (NMDA)-induced fictive locomotion showed pronounced plateau phases. In contrast, crossed caudal (CC) interneurons had a less obvious and steeper plateau region that was followed by a clear notch coinciding with the end of the lateral interneuron plateau phase. These results indicate a significant inhibitory input from LI to CC interneurons.

Animals↗

Interneuron Diversity series: Interneuronal neuropeptides--endogenous regulators of neuronal excitability.

Interneurons are often classified according to neuropeptide content. However, it is becoming increasingly clear that neuropeptides are more than convenient neurochemical markers and can act as important modulators of neuronal activity. Recent advances in understanding neuropeptide release and physiological actions suggest that the interneuronal system of neuropeptides is crucial for maintaining appropriate brain function under normal and pathophysiological conditions. In particular, interneuronal neuropeptides appear to play roles in cognition and as endogenous anti-epileptic agents. This article describes current understanding of the conditions under which neuropeptides are released from interneurons, their specific effects on neuronal excitability and synaptic transmission, and the consequences of their loss of function.

Animals↗

PAX2 is expressed in multiple spinal cord interneurons, including a population of EN1+ interneurons that require PAX6 for their development.

Members of the PAX family of transcription factors are candidates for controlling cell identity in the spinal cord. We have morphologically analyzed cells that express one of these transcription factors, PAX2, demonstrating multiple interneuron cell types express PAX2. Two ventral populations of PAX2-expressing interneurons in the spinal cord are marked by coexpression of the transcription factors, EN1 and EVX1. Interestingly, the expression domains of PAX2, EN1 and EVX1 in postmitotic neurons correlate closely with those of Pax6 and Pax7 in the ventricular zone, implicating these patterning genes in the regulation of PAX2, EN1 and EVX1. We show that one of these patterning genes, Pax6, is required for the correct specification of ventral PAX2+ interneurons that coexpress EN1. These results demonstrate that the early activity of patterning genes in the ventricular zone determines interneuron identity in the spinal cord.

Animals↗

Dual motor output interneurons in the abdominal ganglia of the crayfish Procambarus clarkii: synaptic activation of motor outputs in both the swimmeret and abdominal positioning systems by single interneurons.

Many behavior patterns of the crayfish involve the positioning of the abdomen by the tonic motor system. Movements and positionings of the swimmerets are coordinated with these abdominal movements. Evidence from extracellular analyses suggested that single interneurons of the abdominal nerve cord could produce motor outputs in both the swimmeret and the abdominal positioning systems. Our intracellular investigation has revealed that many single cells can evoke outputs in both motor systems. Interneurons which produced fictive extension or flexion of the abdomen or inhibition of abdominal movement were also able to modulate a variety of swimmeret behavior including cyclic beating and excitation or inhibition of episodic outputs. Although interneurons were discovered that evoked each of the possible classes of dual-output combinations, those that evoked combinations frequently observed in the freely behaving animal were more common than those that evoked infrequently observed combinations. Evidence also indicated that abdominal positioning inhibitors are present in greater numbers than previously suspected and that many are closely associated with the swimmeret circuitry. Interneurons with the ability to start and stop swimmeret cyclic outputs with current injections of opposite polarity are proposed to be higher-order cells, and some are shown to have the properties of trigger neurons. It is proposed that most dual-output cells are presynaptic to single-output cells and that groups of related dual-output cells may function together as command elements.

Animals↗

Characterization of premotor interneurones by their input patterns--application of principal component analysis to cat cervical interneurones.

Principal component analysis of input patterns of cat C6-C8 interneurones (300 cells) revealed that identified premotor interneurones (11 cells) activated from skin afferents and projecting to T1 motoneurones possessed a special input pattern, characterized by restricted distribution on the plane of the first (Prin 1) versus second (Prin 2) principal component (high positive values of both components). These premotor neurones were located mostly in laminae V-VI. Among other laminae V-VI cells descending in the lateral funiculus to T1 similar to such premotor neurones, there were cells distributed similarly on the Prin 1-2 plane. Further, a majority of interneurones antidromically activated from the T1 motor nucleus at low thresholds also showed a distribution on the plane similar to the premotor neurones. We suggest that premotor neurones of this input pattern constitute a major group among laminae V-VI premotor neurones projecting to T1.

Animals↗