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PubMed1996

Inhibition of the respiratory pattern-generating neurons by an identified whole-body withdrawal interneuron of Lymnaea stagnalis

Respiration and the whole-body withdrawal are two incompatible behaviors in the freshwater snail Lymnaea stagnalis. Whole-body withdrawal behavior is believed to be higher on the behavioral hierarchy than respiratory behavior. A central pattern generator (CPG) underlies respiratory behavior; whole-body withdrawal is mediated by a network of electrically coupled neurons. In this study, we provide evidence that the behavioral hierarchy between the whole-body withdrawal and the respiratory behaviors is established at the interneuronal level. We demonstrate that an identified whole-body withdrawal interneuron inhibits both muscular and neuronal components of the respiratory behavior in Lymnaea stagnalis. A pair of identified, electrically coupled interneurons, termed left and right pedal dorsal 11 (L/RPeD11), coordinates the whole-body withdrawal behavior in Lymnaea stagnalis. In the present study, RPeD11 inhibited spontaneously occurring respiratory CPG activity in isolated brain preparations. In addition, electrical stimulation of RPeD11 in a semi-intact preparation also inhibited respiratory CPG interneuron RPeD1. The synaptic connections between RPeD11 and the respiratory CPG neurons RPeD1 and visceral dorsal 4 (VD4) persisted in the presence of high-Ca2+/high-Mg2+ saline, suggesting the possibility that they may be monosynaptic. In a semi-intact preparation (lung­mantle, pneumostome and central nervous system), electrical stimulation of RPeD11 induced pneumostome and columellar muscle contractions while inhibiting the activity of RPeD1. Moreover, mechanical stimulation of the respiratory orifice, the pneumostome, excited RPeD11, while its effects on the respiratory CPG neuron (RPeD1) were inhibitory. To determine the monosynaptic nature of connections between RPeD11 and the respiratory CPG neurons in the intact nervous system, we constructed these synapses in culture. RPeD11 and individual respiratory interneurons were isolated from their respective ganglia and co-cultured under conditions that support neurite outgrowth. Following neuritic overlap, RPeD11 was found to establish inhibitory synapses with the respiratory interneurons, supporting the hypothesis that these synaptic connections are likely to be monosynaptic in the intact central nervous system.

Journal Article↗
PubMed1986

Integrative mechanisms controlling directional sensitivity of an identified sensory interneuron.

Several identified interneurons in the cricket cercal afferent system display directional sensitivity to wind stimuli: the spike frequency of these cells depends on the wind direction with respect to the animal's body. Factors determining the directional sensitivity of one of these identified interneurons (interneuron 10-3) were studied in detail. This cell has 3 dendritic branches that arborize in 3 distinct regions of the terminal abdominal ganglion. Using 2 independent methods, it was demonstrated that the dendrites have different receptive fields to wind stimuli. First, small patches of filiform hairs, whose afferents projected to individual dendrites, were isolated and selectively stimulated. In each case the response of the cell matched the receptive field of the afferents in the patch. Second, a laser beam directed through the stereo dissecting microscope was used to photoinactivate small portions of the cell in situ during intracellular recording. By isolating or ablating individual dendrites, the contributions of each of the 3 dendrites to the overall receptive field were assessed. Although the receptive field of the whole cell could be predicted by a summation of the receptive fields of all 3 dendrites, the precise directional sensitivity of the cell could not be predicted by a simple linear summation of the receptive fields of each dendrite. Two factors were found to account for this nonlinearity of summation. The first factor was polysynaptic inhibition from other interneurons within the terminal abdominal ganglion. Wind directions that activate inhibition in interneuron 10-3 were identified, and the specific classes of filiform afferents that activate the inhibitory pathway were determined. The net effect of the inhibition was to "sharpen" the directional sensitivity of 10-3 by selectively decreasing the cell's response to specific excitatory inputs. The second factor that contributed to directional sensitivity was the complex electroanatomy of the interneuron. The probable location of the spike-initiating zone (SIZ) was determined by using the laser photoinactivation technique. The relative efficacies of synaptic inputs onto the 3 different branches were then interpreted with respect to their different electrotonic distances from the SIZ. On the basis of the data obtained in this report, we present a qualitative model for the basis of directional sensitivity in this cell.

Animals↗
PubMed1998

Quantitative analyses of anatomical and electrotonic structures of crayfish nonspiking interneurons by three-dimensional morphometry.

The three-dimensional structure of premotor nonspiking interneurons in the terminal abdominal ganglion of crayfish have been studied quantitatively by using a confocal laser-scanning microscope. Their passive membrane properties have also been studied electrophysiologically to analyze their electrotonic structure. In either one of the two major morphological types, anterolateral (AL) and posterolateral (PL), that are characterized by different locations of cell bodies in the ganglion, the monopolar cell body is connected with a fine primary process to a thick main segment projecting numerous fine secondary processes. These two types of cells share a common dendritic field in the neuropil, showing similar anatomical characteristics of dendrites. Electrotonic analyses based on the present anatomical and physiological measurements have revealed that the steady-state voltage-attenuation factors for the secondary processes were not statistically different between the AL- and PL-type cells. Comparison between the premotor nonspiking interneurons and an identified sensory nonspiking interneuron, which was studied previously, has revealed that voltage attenuation over secondary processes in both the centripetal and the centrifugal directions was significantly greater in the sensory than in the premotor interneurons, although the anatomical length of each secondary process from its terminal to the main segment was not different between them. Differences in the electrotonic structure between sensory and premotor nonspiking interneurons indicate their different modes of synaptic integration in the control of postsynaptic nerve cells.

Animals↗
PubMed1999

Postnatal development of calretinin- and parvalbumin-positive interneurons in the rat neostriatum: an immunohistochemical study.

On the basis of cytochemical and morphologic differences, two classes of gamma-aminobutyric acidergic (GABAergic) interneurons expressing calcium-binding proteins have been identified in the striatum of adult animals: neurons expressing either parvalbumin (PV) or calretinin (CR). The function of these calcium-binding proteins is not clear, however, they are associated with distinct classes of inhibitory interneurons within the adult neostriatum. By using immunocytochemical techniques, we analyzed the postnatal maturation and the spatiotemporal distribution of PV- and CR-positive neurons in the rat neostriatum compared with a third class of interneurons characterized by the expression of the acetylcholine-synthesizing enzyme, choline acetyltransferase (ChAT). PV-positive cells appeared initially on postnatal day 9 in the lateral region of the striatum. During postnatal weeks 2 and 3, the numbers of PV-positive neurons increased, and this cell population spread progressively in a lateromedial direction. In contrast, CR-expressing neurons were present at birth. During the first few days after birth, the number of CR-immunoreactive cells increased, reaching a peak on postnatal day 5 before declining during the following 2 weeks. A mediolateral gradient was evident temporarily. ChAT-containing neurons were detectable at birth in the lateral striatum. During postnatal weeks 1 and 2, the neurons matured along a lateral-to-medial gradient. The results indicate that the maturation of striatal interneurons is regulated differentially during postnatal development, resulting in a distinct spatiotemporal genesis of phenotypes. The sequential expression of CR and PV suggests a stage-dependent development of subsets of inhibitory interneurons and, hence, the stage-dependent maturation of functionally distinct inhibitory circuits within the neostriatum.

Animals↗
PubMed1996

Premotor interneurons in generation of adaptive leg reflexes and voluntary movements in stick insects.

We investigated the role of local nonspiking interneurons involved in motor control of legs in the stick insects, Carausius morosus. In a preparation that allowed the animals to perform active leg movements such as adaptive tactile reflexes, proprioceptive reflexes, and walking, we gathered the following results. Almost all tested nonspiking interneurons that provide synaptic drive onto moto-neurons of the proximal leg muscles contribute to all of the motor programs underlying tactile reflexes and voluntary leg movements such as walking, searching, and rocking. Most of them are also involved in the generation of proprioceptive reflexes. All motor programs for coactivation, avoidance reflexes, resistance reflexes, and voluntary leg movements result from parallel pathways including nonspiking interneurons that support and others that oppose the motoneuronal activity. The contribution of a single interneuron to the different motor program is specific: it can be supporting for one motor program but opposing for the other. Even for the same motor program, for example, coactivation, the contribution of an individual interneuron can depend on the stimulus site from where the response is elicited. Our results support the idea that the different motor patterns for adaptive tactile reflexes, resistance reflexes, and voluntary leg movements emerge from a multifunctional neuronal circuit that is reorganized corresponding to the motor behavior performed. The actual motor pattern is then shaped by distributed information processing in parallel supporting and opposing pathways.

Acclimatization↗
PubMed2004

Neurogenesis and stereological morphometry of calretinin-immunoreactive GABAergic interneurons of the neostriatum.

We determined the neurogenesis characteristics of a distinct subclass of rat striatum gamma-aminobutyric acidergic (GABAergic) interneurons expressing the calcium-binding protein calretinin (CR). Timed-pregnant rats were given an intraperitoneal injection of 5-bromo-2'-deoxyuridine (BrdU), a marker of cell proliferation, on designated days between embryonic day 12 (E12) and E21. CR-immunoreactive (-IR) neurons and BrdU-positive nuclei were labeled in the adult neostriatum by double immunohistochemistry, and the proportion of double-labeled cells was quantified. CR-IR interneurons of the neostriatum show maximum birth rates (>10% double labeling) between E14 and E17, with a peak at E15. CR-IR interneurons occupying the lateral half of the neostriatum become postmitotic prior to medial neurons. In the precomissural neostriatum, the earliest-born neurons occupy the lateral quadrants and the latest-born neurons occupy the dorsomedial sector. No significant rostrocaudal neurogenesis gradient is observed. CR-IR neurons make up 0.5% of the striatal population and are localized in both the patch and the matrix compartments. CR-IR neurons of the patch compartment are born early (E13-15), with later-born neurons (E16-18) populating mainly the matrix compartment. CR-IR cells of the neostriatum are a distinct subclass of interneurons that are born at an intermediate time during striatal development and share common neurogenesis characteristics with other interneurons and projection neurons produced in the ventral telencephalon.

Age Factors↗
PubMed2001

A confocal study of spinal interneurons in living larval zebrafish.

We used confocal microscopy to examine the morphology of spinal interneurons in living larval zebrafish with the aim of providing a morphological foundation for generating functional hypotheses. Interneurons were retrogradely labeled by injections of fluorescent dextrans into the spinal cord, and the three-dimensional morphology of living cells was reconstructed from confocal optical sections through the transparent fish. At least eight types of interneurons are present in the spinal cord of larval zebrafish; four of these are described here for the first time. The newly discovered cell types include classes of commissural neurons with axons that ascend, descend, and bifurcate in the contralateral spinal cord. Our reexamination of previously described cell types revealed functionally relevant features of their morphology, such as undescribed commissural axons, as well as the relationships between the trajectories of the axons of interneurons and the descending Mauthner axons. In addition to describing neurons, we surveyed their morphology at multiple positions along the spinal cord and found longitudinal changes in their distribution and sizes. For example, some cell types increase in size from rostral to caudal, whereas others decrease. Our observations lead to predictions of the roles of some of these interneurons in motor circuits. These predictions can be tested with the combination of functional imaging, single-cell ablation, and genetic approaches that make zebrafish a powerful model system for studying neuronal circuits.

Animals↗
PubMed2004

Distinct expression of phosphorylated N-methyl-D-aspartate receptor NR1 subunits by projection neurons and interneurons in the striatum of normal and amphetamine-treated rats.

N-methyl-D-aspartate (NMDA) receptors are heteromeric assemblies of subunits (NR1 and NR2A-D), and are enriched in the striatum. Receptor phosphorylation has recently been demonstrated on the NR1 subunit at three serine residues, 897, 896, and 890, which appear to correspond to the level of receptor activity. In this study, expression of phospho-specific NR1 subunits at serine 897 (pNR1S897), serine 896 (pNR1S896), or serine 890 (pNR1S890) in neurochemically identified neurons of the adult rat striatum was detected by using double-immunofluorescent labeling or combined in situ hybridization and immunohistochemistry. In both the dorsal and ventral striatum, pNR1S897 was expressed at high levels in projection neurons containing >55% dynorphin (striatonigral) and >90% enkephalin (striatopallidal) and in interneurons that were 100% positive for choline, >90% positive for parvalbumin, and >45% positive for somatostatin (co-containing neuropeptide Y and neuronal nitric oxide synthase). Low levels of pNR1S896 were present in a small portion of projection neurons (<15% for both populations of projection neurons) and were almost lacking in the three types of interneurons. Interestingly, pNR1S890 was exclusively expressed in most parvalbumin-containing interneurons (70-80%). Acute administration of a psychostimulant, amphetamine, increased the number of dynorphin-containing projection neurons and parvalbumin interneurons showing detectable levels of pNR1S896 and pNR1S890, respectively. These results demonstrate the distinct expression of phospho-NR1 subunits in different populations of striatal projection neurons and interneurons at variable levels in normal rats; they also demonstrate that phosphorylation of NR1, at least on serine 896 and 890 sites, is sensitive to drug exposure.

Amphetamine↗
PubMed2006

Survival of interneurons and parallel fiber synapses in a cerebellar cortex deprived of Purkinje cells: studies in the double mutant mouse Grid2Lc/+;Bax(-/-).

The Lurcher mutation in the Grid2 gene causes the cell autonomous death of virtually all cerebellar Purkinje cells and the target-related death of 90% of the granule cells and 60-75% of the olivary neurons. Inactivation of Bax, a pro-apoptotic gene of the Bcl-2 family, in heterozygous Lurcher mutants (Grid2Lc/+) rescues approximately 60% of the granule cells, but does not rescue Purkinje or olivary neurons. Given the larger size of the cerebellar molecular layer in Grid2Lc/+;Bax(-/-) double mutants compared to Grid2Lc/+ mutants, we analyzed the survival of the stellate and basket interneurons as well as the synaptic connectivity of parallel fibers originating from the surviving granule cells in the absence of their Purkinje cell targets in the Grid2Lc/+;Bax(-/-) cerebellum. Quantification showed a significantly higher density of interneurons ( approximately 60%) in the molecular layer of the Grid2Lc/+;Bax(-/-) mice compared to Grid2Lc/+, suggesting that interneurons are subject to a BAX-dependent target-related death in the Lurcher mutants. Furthermore, electron microscopy showed the normal ultrastructural aspect of a number of parallel fibers in the molecular layer of the Grid2Lc/+; Bax(-/-) double mutant mice and preserved their numerous synaptic contacts on interneurons, suggesting that interneurons could play a trophic role for axon terminals of surviving granule cells. Finally, parallel fibers varicosities in the double mutant established "pseudo-synapses" on glia as well as displayed autophagic profiles, suggesting that the connections established by the parallel fibers in the absence of their Purkinje cell targets were subject to a high turnover involving autophagy.

Animals↗
PubMed1975

Deafferentation slows the growth of specific dendrites of identified giant interneurons.

The effect deafferentation has on the morphology of giant interneurons was studied in the abdominal nervous system of crickets (Acheta domesticus). The morphology of four uniquely identified giant interneurons was exxamined by iontophoresing cobalt chloride into the neurons of interest. A major source of afferents for these interneurons consists of mechanoreceptors located on paired abdominal sensory appendages -- the cerci. Partial deafferentation of the giant interneurons was obtained by pinching off the cercus at hatching and maintaing the specimen in this deprived condition until adulthood. The interneurons of three groups of animals were examined; control specimens which were not treated surgically, unilaterally treated specimens which had a single cercus removed and bilaterally treated specimens which had both cerci removed. Two types of morphological changes were detected. (1) Chronic removal of a cercus was correlated with a reduction in length of dendrites ipsilateral to the ablated cercus; however, the general form of the dendritic branching pattern remained constant and recognizable. Two dendrites of a single neuron could be influenced independently if they were innervated by separate cerci. Thus deprivation did not have a generalized effect on growth of a neuron, rather it specifically influenced the dendrites deprived of afferents. (2) It was also observed that the projection of cercal sensory fibers in specimens reared with a single cercus differed from normal in that scattered fibers cross the midline in regions of the ganglion where none usually exist. It is suggested that modifications in the response properties of these deprived neurons are based on these two changes in morphology.

Animals↗
PubMed1983

The morphology, ultrastructure, and distribution of synapses on an intersegmental interneuron of the locust.

The structure and synaptic relationships of an intersegmental interneurone in the mesothoracic ganglion of the locust were studied by intracellular labelling for light and electron microscopy. Before labelling, the interneurone was characterized physiologically. It is depolarized and produces spikes when either of the two forewings are elevated, and by sensory inputs from the tarsi of the middle legs. The gross morphology of the interneurone is quite distinctive. Its cell body is on the ventral surface of the ganglion near the midline and its primary neurite lies in the T-tract. Its axon performs a characteristic lateral loop around the lateral dorsal tract before entering an anterior connective and ascending at least as far as the prothoracic ganglion. Within the mesothoracic ganglion, the interneurone gives rise to a profusion of branches. Ultrastructural examination and reconstruction of three regions of the interneurone from serial sections shows that input and output synapses are present in approximately equal numbers, and often are close to each other. There is no evidence for separate input and output regions. Output synapses are characterized by the presence of a presynaptic bar and a population of 500-2,000 round, electron-lucent vesicles. Most of the estimated 10(5)-10(6) synapses occur on the smaller diameter processes or on spines. The spines are of uniform diameter along their lengths, and are pre- or postsynaptic or both, but the postsynaptic spines are of smaller diameter than the others.

Animals↗
PubMed1992

Morphology of intracellularly labeled interneurons in the dentate gyrus of the immature rat.

Although many aspects of the morphological development of interneurons in the dentate gyrus have been described, the full extent of their dendrites and local axon projections in immature rodents has not been examined. Here intracellular labeling was used to assess the branching patterns of interneurons in the dentate gyrus of rat pups between 7 and 9 days of age. Labeled neurons were located within or just below the granule cell layer, and most were classified as GABAergic basket neurons on the basis of their dendritic morphologies. All labeled interneurons exhibited immature characteristics. Spines were present on cell bodies and dendrites, and growth cones were visible on some dendrites and axons. In spite of these immature features, the dendrites and axon arbors of the labeled neurons were extensive. Many apical dendrites reached the top of the molecular layer, and a number of basal dendrites extended to the CA3 pyramidal cell layer of the hippocampus. Elaborate axon plexuses were present within the dentate gyrus itself, and axon collaterals of several neurons extended beyond the dentate gyrus to branch within regions CA3 and CA1 of the hippocampus. These results indicate that the dendrites and axon collaterals of dentate interneurons are extensive at a time when the principal neurons, the granule cells, are still proliferating. These data are consistent with the idea that GABAergic interneurons may influence granule cell development in the dentate gyrus, as well as pyramidal cell maturation in the hippocampus proper.

Animals↗
PubMed1994

Spinal cord lamina V and lamina VII interneuronal projections to sympathetic preganglionic neurons.

This light and electron microscopic study sought to localize spinal cord interneurons that contribute to the normal and abnormal physiological regulation of spinal sympathetic preganglionic function. Sympathetic preganglionic neurons in caudal C8 through T4 of rat spinal cord were retrogradely labeled with wheat germ agglutinin (WGA) and/or cholera beta subunit (CT beta) following injections into the superior cervical ganglion (SCG). With two exceptions, the observed locations of retrogradely WGA- and CT beta-labeled sympathetic preganglionic neurons were as expected from previous studies. The exceptions were restricted populations of cells in caudal C8 and rostral T1 spinal segments. These neurons were classified as ventrolateral (vlSPN) and ventromedial (vmSPN) sympathetic preganglionic neurons; their somata and dendrites encircled dorsolateral lamina IX motoneurons. Only WGA was transported transneuronally following the retrograde labeling of sympathetic preganglionic neurons. Transneuronally WGA-labeled spinal interneurons were located principally in the reticulated division of lamina V and dorsolateral lamina VII. A strict segmental organization was observed. All transneuronally labeled interneurons were ipsilateral to, and coextensive with, retrogradely WGA-labeled sympathetic preganglionic neurons. Electron microscopic observations suggested that retrograde transsynaptic passage of WGA occurred within the sympathetic preganglionic neuropil and showed further that similar classes of organelles were WGA immunoreactive in retrogradely labeled sympathetic preganglionic neurons and in transneuronally labeled lamina V and lamina VII neurons: 1) cisternae and vesicles at the trans face of the Golgi apparatus, 2) large endosomes/dense bodies, and 3) multivesicular bodies. The data are consistent with two hypotheses: 1) Somatic and visceral primary afferent inputs to thoracic spinal cord modify segmental sympathetic preganglionic function through activation of a disynaptic pathway involving lamina V and/or lamina VII interneurons, and 2) long-loop propriospinal pathways access sympathetic preganglionic neurons through symmetrical, segmental interneuronal circuitry.

Animals↗
PubMed1994

Responses of thoracic interneurons to tactile stimulation in cockroach, Periplaneta americana.

Recent findings indicate that cockroaches escape in response to tactile stimulation as well as they do in response to the classic wind puff stimulus. The thoracic interneurons that receive inputs from ventral giant interneurons also respond to tactile stimulation and, therefore, represent a potential site of convergence between wind and tactile stimulation, as well as other sensory modalities. In this article, we characterize the tactile response of these interneurons, which are referred to as type-A thoracic interneurons (TIAs). In response to tactile stimulation of the body cuticle, TIAs typically respond with a short latency biphasic depolarization which often passes threshold for action potentials. The biphasic response is not typical of responses to wind stimulation nor of tactile stimulation of the antennae. It is also not seen in tactile responses of thoracic interneurons that are not part of the TIA group. The responses of individual TIAs to stimulation of various body locations were mapped. The left-right directional properties of TIAs are consistent with their responses to wind puffs from various different directions. Cells that respond equally well to wind from the left and right side also respond equally well to tactile stimuli on the left and right side of the animal's body. In contrast, cells that are biased to wind on one side are also biased to tactile stimulation on the same side. In general, tactile responses directed at body cuticle are phasic rather than tonic, occurring both when the tactile stimulator is depressed and released. The response reflects stimulus strength and follows repeated stimulation quite well. However, the first phase of the biphasic response is more robust during high-frequency stimulation than the second phase. TIAs also respond to antennal stimulation. However, here the response characteristics are complicated by the fact that movement of either antenna evokes descending activity in both left and right thoracic connectives. The data suggest that the TIAs make up a multimodal site of sensory convergence that is capable of generating an oriented escape turn in response to any one of several sensory cues.

Animals↗
PubMed1995

Dual pathways for tactile sensory information to thoracic interneurons in the cockroach.

The escape system of the American cockroach is both fast and directional. In response to wind stimulation both of these characteristics are largely due to the properties of the ventral giant interneurons (vGIs), which conduct sensory information from the cerci on the rear of the animal to type A thoracic interneurons (TIAs) in the thoracic ganglia. The cockroach also escapes from tactile stimuli, and although vGIs are not involved in tactile-mediated escapes, the same thoracic interneurons process tactile sensory information. The response of TIAs to tactile information is typically biphasic. A rapid initial depolarization is followed by a longer latency depolarization that encodes most if not all of the directional information in the tactile stimulus. We report here that the biphasic response of TIAs to tactile stimulation is caused by two separate conducting pathways from the point of stimulation to the thoracic ganglia. Phase 1 is generated by mechanical conduction along the animal's body cuticle or other physical structures. It cannot be eliminated by complete lesion of the nerve cord, and it is not evoked in response to electrical stimulation of abdominal nerves that contain the axons of sensory receptors in abdominal segments. However, it can be eliminated by lesioning the abdominal nerve cord and nerve 7 of the metathoracic ganglion together, suggesting that the relevant sensory structures send axons in nerve 7 and abdominal nerves of anterior abdominal ganglia. Phase 2 of the TIA tactile response is generated by a typical neural pathway that includes mechanoreceptors in each abdominal segment, which project to interneurons with axons in either abdominal connective. Those interneurons with inputs from receptors that are ipsilateral to their axon have a greater influence on TIAs than those that receive inputs from the contralateral side. The phase 1 response has an important role in reducing initiation time for the escape response. Animals in which the phase 2 pathway has been eliminated by lesion of the abdominal nerve cord are still capable of generating a partial startle response with a typically short latency even when stimulated posterior to the lesion.

Animals↗
PubMed1995

Protein kinase C inhibitors enhance the 5-HT2A receptor-mediated excitatory effects of serotonin on interneurons in rat piriform cortex.

Previously it has been shown that excitatory effects of 5-hydroxytryptamine (5-HT) upon interneurons in the rat piriform cortex are mediated by 5-HT2A receptors. This receptor is linked to phosphoinositide turnover, and one consequence of stimulating this receptor is the activation of protein kinase C (PKC). In the present study, the effect of PKC inhibitors on the 5-HT excitation of piriform cortical interneurons was examined by extracellular recording in a rat brain slice preparation. Bath application of the selective PKC inhibitors, bisindolymalemide and chelerythrine, and the nonselective protein kinase inhibitor, H-7, all enhanced the excitatory effects of 5-HT. Two other nonselective protein kinase inhibitors, H-8 and HA 1004, which are 2.5-fold and 6.7-fold less potent than H-7 at inhibiting PKC, produced a slight or no enhancement, respectively, of the excitatory effect of 5-HT. Bisindolylmalemide, chelerytrine, and H-7 did not enhance the excitatory effects of norepinephrine or carbachol on the same interneurons. The PKC activator phorbol 12, 13-diacetate (PDA) decreased the excitatory effect of 5-HT; this decrease was rapidly reversed by H-7. As inhibitors of PKC selectively enhanced rather than blocked the excitation by 5-HT mediated by 5-HT2A receptors, we conclude that activation of PKC does not mediate the excitation by 5-HT of piriform cortical interneurons. Instead, we propose that PKC may have a negative feedback role in modulating the excitation by 5-HT of piriform cortical interneurons.

Animals↗
PubMed2002

Slit promotes branching and elongation of neurites of interneurons but not projection neurons from the developing telencephalon.

Proper neuronal migration and establishment of circuitry are key processes for laying down the functional network of cortical neurons. A variety of environmental guidance cues, attractive or repulsive, have been shown to guide cell migration and axon arborization. One of these, Slit, appears to possess contrarian properties; it can either inhibit axon outgrowth or promote branching and elongation. The object of the present study was to assess the effect of Slit on MGE and neocortical neurons in culture and in the developing ventricle. When cocultured with a Slit source, E13.5 MGE explants displayed inhibited neurite outgrowth while GABA neuron dispersion away from Slit was increased. Similar inhibition of neurite outgrowth was seen in dissociated cells from E13.5 MGE, these cells were identified to be interneurons based upon their GABA staining. In contrast, E13.5 interneurons, after culture for another 5 days, were responsive to Slit by neurite branching and elongation. Projection neurons, identified by lack of GABA staining, did not respond to Slit, either by branching or elongation. Furthermore, GABA interneurons but not pyramidal neurons, appeared to avoid neocortical areas close to an implanted source of Slit in the ventricular wall. These results lead us to suggest that interneurons but not projection neurons are responsive to the chemorepellant effect of Slit. However, more mature interneurons appear to respond to Slit by neurite arborization. These results demonstrate a selective response to Slit by GABAergic neurons during neocortical development.

Animals↗
PubMed1994

A persistent sodium current contributes to oscillatory activity in heart interneurons of the medicinal leech.

1. Normal activity in bilateral pairs of heart interneurons, from ganglia 3 or 4, in the medicinal leech (Hirudo medicinalis) is antiphasic due to their reciprocally inhibitory connections. However, Ca(++)-free Co(++)-containing salines lead to synchronous oscillations in these neurons. 2. Internal TEA+ allows expression of full plateaus during Co++ induced oscillations in heart interneurons; these plateaus are not blocked by Cs+. Similar plateaus are also observed with internal TEA+ alone, but under these conditions activity in heart interneurons from ganglia 3 or 4 is antiphasic. 3. Plateaus in heart interneurons induced by Co++ and internal TEA+ involve a conductance increase. 4. A voltage-dependent inward current, IP, showing little inactivation, was isolated using single-electrode voltage-clamp in heart interneurons. This current is carried at least in part by Na+; the current is reduced when external Na+ is reduced and is carried by Li++ when substituted for Na+. 5. Calcium channel blockers such as La3+ and Co++ block neither the TEA+ induced plateaus nor IP, suggesting that Na+ is not using Ca++ channels. Moreover, IP is enhanced by Ca(++)-free CO(++)-containing salines. Thus, IP is correlated with the TEA(+)- and Co(++)-induced plateau behavior.

Animals↗