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PubMed2001

Opposing actions of nitric oxide on synaptic inputs of identified interneurones in the central nervous system of the crayfish.

Little is known of the action of nitric oxide (NO) at the synaptic level on identified interneurones in local circuits that process mechanosensory signals. Here, we examine the action of NO in the terminal abdominal ganglion of the crayfish Pacifastacus leniusculus, where it has modulatory effects on the synaptic inputs of 17 identified ascending interneurones mediated by electrical stimulation of a sensory nerve. To analyse the role of NO in the processing of sensory signals, we bath-applied the NO donor SNAP, the NO scavenger PTIO, the nitric oxide synthase (NOS) inhibitor l-NAME, the NOS substrate l-arginine, a cyclic GMP (cGMP) analogue, 8-Br-cGMP, and the soluble guanylate cyclase (sGC) inhibitor ODQ. The effects of these chemicals on the synaptic inputs of the interneurones could be divided into two distinct classes. The NO donor SNAP enhanced the inputs to one class of interneurone (class 1) and depressed those to another (class 2). Neither the inactive isomer NAP nor degassed SNAP had any effect on the inputs to these same classes of interneurone. The NO scavenger PTIO caused the opposite effects to those of the NO donor SNAP, indicating that endogenous NO may have an action in local circuits. Preventing the synthesis of NO using l-NAME had the opposite effect to that of SNAP on each response class of interneurone. Increasing the synthesis of endogenous NO by applying l-arginine led to effects on both response classes of interneurone similar to those of SNAP. Taken together, these results suggested that NO was the active component in mediating the changes in amplitude of the excitatory postsynaptic potentials. Finally, the effects of 8-Br-cGMP were similar to those of the NO donor, indicating the possible involvement of a NO-sensitive guanylate cyclase. This was confirmed by preventing the synthesis of cGMP by sGC using ODQ, which caused the opposite effects to those of 8-Br-cGMP on the two response classes of interneurone. The results indicate that a NO--cGMP signal transduction pathway, in which NO regulates transmitter release from mechanosensory afferents onto intersegmental ascending interneurones, is probably present in the local circuits of the crayfish.

Animals↗
PubMed2002

Serotonergic modulation of nonspiking local interneurones in the terminal abdominal ganglion of the crayfish.

The modulatory effect of serotonin on local circuit neurones forming the uropod motor control system of the crayfish Procambarus clarkii Girard was analysed electrophysiologically. Bath application of 10 micro mol l(-1) serotonin caused a decrease in the tonic spike activity of the exopodite reductor motor neurone. The inhibitory effect of serotonin on the motor neurone was dose-dependent and its spike discharge was completely suppressed for long periods by 1 mmol l(-1) serotonin perfusion. Nonspiking local interneurones in the terminal abdominal ganglion showed either a membrane depolarization (N=6) or hyperpolarization (N=9) of 10-30 mV in amplitude when 100 micro mol l(-1) serotonin was perfused for 3-5 min. By contrast, spiking local interneurones and intersegmental ascending interneurones showed no observable excitatory responses to the perfusion of serotonin but instead some showed a small membrane hyperpolarization of 2-5 mV. These results indicate that the nonspiking interneurones could contribute substantially to the level of tonic excitation of the uropod motor neurones. Sensory stimulation elicited depolarizing or hyperpolarizing potentials in the nonspiking interneurones and excitatory postsynaptic potentials (EPSPs) and spikes in the spiking interneurones. The sensory responses of spiking interneurones increased during bath application of serotonin and were reduced after 20-30 min of washing with normal saline. In the nonspiking interneurones, the amplitude of both depolarizing and hyperpolarizing potentials increased without any direct correlation with the serotonin-mediated potential change. This effect of serotonin was long-lasting and continued to enhance the responses of the nonspiking interneurones after washing. This postserotonin enhancement persisted for over 1 h.

Abdomen↗
PubMed2002

Control of feedforward dendritic inhibition by NMDA receptor-dependent spike timing in hippocampal interneurons.

Two putative functional populations of feedforward interneurons with distinct spike-timing properties were identified in stratum radiatum of the CA1 rat hippocampus. Interneurons with fast (half width, <100 msec) EPSPs fired after short EPSP-spike latencies and with a high degree of temporal precision compared with cells with slow (half width, >100 msec) EPSPs. Spike timing in fast and slow interneurons occurred at different phases of the EPSPs of simultaneously activated pyramidal cells. In addition, firing of fast interneurons preceded action potentials in principal neurons, whereas action potentials in slow interneurons could either precede or follow firing in pyramidal cells. Temporal integration of separate inputs leading to synaptically evoked firing was more prominent in slow than fast interneurons. Functional diversity between the two putative populations was abolished by the NMDA receptor (NMDAR) antagonist d-(-)-2-amino-5-phosphonopentanoic acid (d-AP-5). The axon of both cell types was primarily restricted to striatum radiatum or to striatum lacunosum-moleculare in the case of slow cells, suggesting targeting of principal cell dendrites for the majority of the cells of this study. Indeed, firing of slow and fast interneurons generated similar unitary IPSCs (uIPSCs) in pyramidal neurons. uIPSCs were mediated by GABA(A) receptors and had in most cases small amplitudes and slow kinetics. Our results suggest that functionally heterogeneous interneurons encode the temporal properties of dendritic feedforward inhibition, and that NMDARs play an essential role in shaping the integrative properties of interneurons and in setting the timing of GABA release.

2-Amino-5-phosphonovalerate↗
PubMed2002

Modular organization of turtle spinal interneurons during normal and deletion fictive rostral scratching.

During normal rostral scratching in the spinal turtle, there is rhythmic alternation between hip-flexor and hip-extensor motor activity. During rostral scratching with hip-extensor deletions, there are successive bursts of hip-flexor motor activity and no activity in hip-extensor motor neurons. We characterized the ON- and OFF-phases of 72 descending propriospinal interneurons with distinct activity bursts during normal rostral scratching. We also studied the activity of these interneurons during deletion scratching. Hip-extensor interneurons were active when hip-flexor motor neurons were quiet in normal scratching and had zero overlap with hip-flexor motor activity. This population of hip-extensor interneurons, termed the hip-extensor module or hip-extensor unit-burst generator, was mainly quiet during deletion scratching. Our observation supports the concept that a module is a neuronal population that may be active or quiet in a coordinated manner during a spinal motor rhythm. During normal scratching, hip-flexor interneurons were active during hip-flexor motor activity, and spanning interneurons were active during both hip-flexor motor activity and quiescence. Hip-flexor and spanning interneurons with intermediate overlap with hip-flexor motor activity fired in bursts during deletion scratching. Hip-flexor and spanning interneurons with large overlap with hip-flexor motor activity fired continuously during deletion scratching. Key features of hip-flexor and spanning interneuron firing during normal scratching were preserved during deletion scratching. Thus these features do not require activity in the hip-extensor module in every cycle of a motor rhythm.

Action Potentials↗
PubMed1999

Identification of an interneuronal population that mediates recurrent inhibition of motoneurons in the developing chick spinal cord.

Studies on the development of synaptic specificity, embryonic activity, and neuronal specification in the spinal cord have all been limited by the absence of a functionally identified interneuron class (defined by its unique set of connections). Here, we identify an interneuron population in the embryonic chick spinal cord that appears to be the avian equivalent of the mammalian Renshaw cell (R-interneurons). These cells receive monosynaptic nicotinic, cholinergic input from motoneuron recurrent collaterals. They make predominately GABAergic connections back onto motoneurons and to other R-interneurons but project rarely to other spinal interneurons. The similarity between the connections of the developing R-interneuron, shortly after circuit formation, and the mature mammalian Renshaw cell raises the possibility that R-interneuronal connections are formed precisely from the onset. Using a newly developed optical approach, we identified the location of R-interneurons in a column, dorsomedial to the motor nucleus. Functional characterization of the R-interneuron population provides the basis for analyses that have so far only been possible for motoneurons.

Animals↗
PubMed2001

Diverse types of interneurons generate thalamus-evoked feedforward inhibition in the mouse barrel cortex.

Sensory information, relayed through the thalamus, arrives in the neocortex as excitatory input, but rapidly induces strong disynaptic inhibition that constrains the cortical flow of excitation both spatially and temporally. This feedforward inhibition is generated by intracortical interneurons whose precise identity and properties were not known. To characterize interneurons generating feedforward inhibition, neurons in layers IV and V of mouse somatosensory ("barrel") cortex in vitro were tested in the cell-attached configuration for thalamocortically induced firing and in the whole-cell mode for synaptic responses. Identification as inhibitory or excitatory neurons was based on intrinsic firing patterns and on morphology revealed by intracellular staining. Thalamocortical stimulation evoked action potentials in approximately 60% of inhibitory interneurons but in <5% of excitatory neurons. The inhibitory interneurons that fired received fivefold larger thalamocortical inputs compared with nonfiring inhibitory or excitatory neurons. Thalamocortically evoked spikes in inhibitory interneurons followed at short latency the onset of excitatory monosynaptic responses in the same cells and slightly preceded the onset of inhibitory responses in nearby neurons, indicating their involvement in disynaptic inhibition. Both nonadapting (fast-spiking) and adapting (regular-spiking) inhibitory interneurons fired on thalamocortical stimulation, as did interneurons expressing parvalbumin, calbindin, or neither calcium-binding protein. Morphological analysis revealed that some interneurons might generate feedforward inhibition within their own layer IV barrel, whereas others may convey inhibition to upper layers, within their own or in adjacent columns. We conclude that feedforward inhibition is generated by diverse classes of interneurons, possibly serving different roles in the processing of incoming sensory information.

Action Potentials↗
PubMed2001

Focal inhibitory interneuron loss and principal cell hyperexcitability in the rat hippocampus after microinjection of a neurotoxic conjugate of saporin and a peptidase-resistant analog of Substance P.

Episodes of prolonged seizures or head trauma produce chronic hippocampal network hyperexcitability hypothesized to result primarily from inhibitory interneuron loss or dysfunction. The possibly causal role of inhibitory neuron failure in the development of epileptiform pathophysiology remains unclear because global neurologic injuries produce such a multitude of effects. The recent finding that Substance P receptors (SPRs) are expressed exclusively in the rat hippocampus by inhibitory interneurons provided the rationale for attempting to ablate interneurons selectively by using neurotoxic conjugates of SPR ligands and the ribosome inactivating protein saporin that specifically target Substance P receptor-expressing cells. Whereas intrahippocampal microinjection of a conjugate of native SP and saporin produced significant nonspecific damage at concentrations needed to produce even limited selective loss of SPR-positive cells, a conjugate of saporin and the more potent and peptidase-resistant SP analog [Sar(9), Met(O(2))(11)] Substance P (SSP-saporin) caused negligible nonspecific damage at the injection site, and a virtually complete loss of SPR-like immunoreactivity (LI) up to 1 mm from the injection site. Within the SPR depletion zone, immunoreactivities for most GABA-, parvalbumin-, somatostatin-, and cholecystokinin-immunoreactive cells and fibers were eliminated. The few interneurons detectable within the affected zone were devoid of SPR-LI. The apparent loss of interneurons was selective in that calbindin- and glutamate receptor subunit 2 (GluR2) -positive principal cells survived within the affected zone, as did myelinated fibers and the extrinsic calretinin- and tyrosine hydroxylase--immunoreactive terminals of subcortical afferents. An apparent lack of reactive synaptic reorganization in response to interneuron loss was indicated by zinc transporter-3 (ZnT3)-- and beta-synuclein--LI, as well as by Timm staining, all of which revealed relatively normal patterns of excitatory terminal distribution. Control injections produced minor damage at the injection site, but no apparent specific loss of SPR-LI. One to 12 weeks after injection of SSP-saporin, extracellular electrophysiological field responses recorded in the CA1 pyramidal and dentate granule cell layers in response to afferent stimulation were blindly evaluated simultaneously in two sites 1-2 mm apart along the longitudinal hippocampal axis. SSP-saporin-treated rats exhibited relatively normal responses in some sites, whereas disinhibition and hyperexcitability indistinguishable from the pathophysiology produced by experimental status epilepticus were simultaneously recorded at adjacent sites. Anatomic analysis of the recording sites in each animal revealed that epileptiform pathophysiology was consistently observed only within areas of SPR ablation, whereas relatively normal evoked responses were recorded from immediately adjacent and relatively unaffected regions. These data establish the efficacy of [Sar(9), Met(O(2))(11)] Substance P-saporin for producing a selective and spatially extensive ablation of hippocampal inhibitory interneurons in vivo and a highly focal disinhibition that was restricted to the site of interneuron loss. These results also demonstrate that the "epileptic" pathophysiology produced by experimental status epilepticus or head trauma can be replicated by focal interneuron loss per se, without involving principal cell loss and other interpretive confounds inherent in the use of global neurologic injury models.

Animals↗
PubMed2002

Motoneuron-derived neurotrophin-3 is a survival factor for PAX2-expressing spinal interneurons.

Rat spinal cord interneurons undergo programmed cell death shortly after birth. We investigated here whether cell death of interneurons could be regulated by trophic factors produced by motoneurons, one of their main targets. To test this hypothesis, we studied the effect of the selective destruction of motoneurons on the survival of interneurons in organotypic cultures of embryonic rat spinal cords. Motoneurons were eliminated by an anti-p75(NTR)-specific immunotoxin (192 IgG-saporin). We then observed a decrease of 28% in the number of ventral spinal interneurons immunoreactive (IR) for the homeoprotein PAX2. This was correlated with an increase in the number of apoptotic nuclei in the same area. Because neurotrophin-3 (NT-3) is specifically produced by motoneurons and because interneurons express the NT-3 high-affinity receptor trkC, we examined the role of NT-3 in the survival of PAX2-IR interneurons. Addition of NT-3 to 192 IgG-saporin-treated explants rescued ventral PAX2-IR interneurons. Depletion of secreted NT-3 by anti-NT-3 antibodies induced 66% loss of ventral PAX2-IR interneurons. We conclude that motoneuron-derived NT-3 is a trophic factor for ventral PAX2-IR interneurons.

Animals↗
PubMed2003

Genetic disruption of cortical interneuron development causes region- and GABA cell type-specific deficits, epilepsy, and behavioral dysfunction.

The generation of properly functioning circuits during brain development requires precise timing of cell migration and differentiation. Disruptions in the developmental plan may lead to neurological and psychiatric disorders. Neocortical circuits rely on inhibitory GABAergic interneurons, the majority of which migrate from subcortical sources. We have shown that the pleiotropic molecule hepatocyte growth factor/scatter factor (HGF/SF) mediates interneuron migration. Mice with a targeted mutation of the gene encoding urokinase plasminogen activator receptor (uPAR), a key component in HGF/SF activation and function, have decreased levels of HGF/SF and a 50% reduction in neocortical GABAergic interneurons at embryonic and perinatal ages. Disruption of interneuron development leads to early lethality in most models. Thus, the long-term consequences of such perturbations are unknown. Mice of the uPAR-/- strain survive until adulthood, and behavior testing demonstrates that they have an increased anxiety state. The uPAR-/- strain also exhibits spontaneous seizure activity and higher susceptibility to pharmacologically induced convulsions. The neocortex of the adult uPAR-/- mouse exhibits a dramatic region- and subtype-specific decrease in GABA-immunoreactive interneurons. Anterior cingulate and parietal cortical areas contain 50% fewer GABAergic interneurons compared with wild-type littermates. However, interneuron numbers in piriform and visual cortical areas do not differ from those of normal mice. Characterization of interneuron subpopulations reveals a near complete loss of the parvalbumin subtype, with other subclasses remaining intact. These data demonstrate that a single gene mutation can selectively alter the development of cortical interneurons in a region- and cell subtype-specific manner, with deficits leading to long-lasting changes in circuit organization and behavior.

Animals↗
PubMed1992

Single local interneurons in the locust make central synapses with different properties of transmitter release on distinct postsynaptic neurons.

Quantal analysis has been applied to the inhibitory synapses made by single spiking local interneurons onto several nonspiking local interneurons (and motorneurons) in the locust CNS. Transmission at these synapses appears to be mediated by GABA. The apparent reversal potential of the IPSP and inhibitory postsynaptic current were -80 to -85 mV, a value similar to that of the potential evoked by pressure-applied GABA. This reversal potential was 25-30 mV more negative than the resting potential of the nonspiking interneurons in the experimental conditions. The statistical properties of release at these synapses were studied by recording simultaneously from pre- and postsynaptic interneurons with intracellular electrodes. The distribution of postsynaptic potential amplitudes could be described by a simple binomial model, implying uniformity of binomial p (probability of release at a single release site) for each synapse. The mean quantal amplitude was 290 +/- 110 microV, and the mean quantal content m of the IPSPs was 6.25 +/- 2.83. The mean values of binomial n (average size of the releasable pool) and p were 13.11 +/- 2.8 and 0.45 +/- 0.16, respectively. Numerical simulations of statistical experiments were performed to test whether the IPSP amplitude distribution histograms might be misleadingly indicative of quantal release. These simulations showed that such a hypothesis was very unlikely. When a spiking local interneuron was impaled, several of its target interneurons could sometimes be successively sampled. Quantal analysis was then performed with the different IPSPs evoked, in identical conditions, by a same presynaptic interneuron, and the quantal parameters were compared between the synapses. It was found that binomials n and p and their product m generally differed between the synapses made by a given spiking interneuron onto different target neurons. These results show that quantal contents can vary for the many synapses made centrally by one interneuron, and suggest that this variability may arise from differences in release probabilities between the sites associated with different synapses.

Animals↗
PubMed2004

[Specification of dorsal interneurons by Wnt signaling].

During development of the vertebrate central nervous system (CNS), highly proliferative cells in the ventricular zone of the neural tube serve themselves as progenitors of the various types of neurons, such as interneurons and motor neurons. In the dorsal half of the spinal cord, signals from the roof plate are required for the development of three classes of dorsal interneuron; D1, D2 and D3, listed from dorsal to ventral. This study demonstrates that mouse embryos lacking both Wnt-1 and Wnt-3a are indeed defective in determination of dorsal interneurons. Generation of D1 and D2 classes of dorsal interneurons is impaired; this loss of the dorsal interneurons is compensated by a dorsal expansion of D3 interneuron populations. Most importantly, expression of BMP family proteins, which have been reported to be able to induce dorsal interneurons, is not significantly affected in these mutant embryos. Moreover, the induction of D1 and D2 class interneurons by Wnt-3a protein in the isolated medial region of chick neural plate is demonstrated. Together, these observations clearly indicate that Wnt signaling has a critical role in the generation of D1 and D2 dorsal interneurons.

Animals↗
PubMed1993

Response dynamics and directional properties of nonspiking local interneurons in the cockroach cercal system.

The response properties and directional receptive fields of nonspiking local interneurons in the cercal system of the cockroach are described. Wind-evoked responses were recorded intracellularly, and then analyzed by means of the Wiener kernel method in which a Gaussian white noise signal was used as a stimulus. Cross-correlation between the response and the white noise signal produced first- (linear) and second-order (nonlinear) kernels that were used to define input-output characteristics of the interneurons. Three sets of interneurons were distinguished on the basis of kernel analysis. First, responses in interneurons 101, 107, 111, and 203 were characterized predominantly by a differentiating first-order kernel, which suggests a linear relationship to the stimulus. The amplitude and waveform of the kernel changed with the change in stimulus angle, indicating that these four cells are directionally sensitive. Second, responses in interneurons 102 and 103 were also directionally sensitive but highly nonlinear. The first-order kernel was biphasic, whereas the second-order kernel had an elongated depolarizing peak on the diagonal. The response dynamics were accounted for by a cascade of two filters, a linear band-pass filter and a static nonlinear filter, in which the nonlinearity is a signal compression (or a rectification). Third, responses in interneurons 104 and 201 consist largely of the second-order nonlinear component. The second-order kernel, which had an elongated depolarizing peak or a hyperpolarizing valley on the diagonal, did not show any directional preference. The second-order nonlinearity was dynamic, and could be modeled by a band-pass linear filter-static nonlinearity-low-pass linear filter cascade, where the static nonlinearity is a full-wave rectification. The band-pass filter would simply reflect the mechanical property of cercal hair sensilla, whereas the low-pass filter represents the transfer at synapses between the cercal afferents and the interneurons. The nonlinear response thus explains the difference in the directional sensitivity while the differentiating first-order kernel explains the velocity sensitivity of the interneurons. We show that 101 and 107 respond most preferentially to wind from the left versus right, whereas 102, 103, 111, and 203 respond to wind from the front versus rear. Thus, it is suggested that there are two subsystems responding maximally to wind displacement along two coordinate directions, one for the longitudinal direction and the other for the transverse direction. On the other hand, the full-wave-rectifier nonlinear interneurons are omnidirectional, and thus suggested to code simply the power of the wind displacement.

Abdomen↗
PubMed1997

Interneurons of the dentate-hilus border of the rat dentate gyrus: morphological and electrophysiological heterogeneity.

Interneurons located near the border of the dentate granule cell layer and the hilus were studied in hippocampal slices using whole-cell current clamp and biocytin staining. Because these interneurons exhibit both morphological and electrophysiological diversity, we asked whether passive electrotonic parameters or repetitive firing behavior correlated with axonal distribution. Each interneuron was distinguished by a preferred axonal distribution in the molecular layer or granule cell layer, and four groups could be discerned, the axons of which arborized in (1) the granule cell layer, (2) the inner molecular layer, (3) the outer molecular layer, and (4) diffusely in the molecular layer. In our sample, interneurons with axons arborizing diffusely in the molecular layer were most frequent, and those with axons restricted to the granule cell layer were least frequent. Resting potential, input resistance, time constant, electrotonic length, and spike frequency adaptation (SFA) were not significantly different among the four groups, and the variability in SFA between cells with similar axonal distributions was striking. Clear differences in action potential morphology and afterhyperpolarizations, however, emerged when nonadapting interneurons were compared with those exhibiting SFA. Interneurons exhibiting SFA had characteristically broader spikes, progressive slowing of action potential repolarization during repetitive firing, and slow afterhyperpolarizations that distinguished them from nonadapting interneurons. We propose that the variability in repetitive firing behavior and morphology exhibited by each of these interneurons makes each interneuron unique and may provide a high level of fine tuning of inhibitory control critical to information processing in the dentate.

Action Potentials↗
PubMed1996

Interneurones involved in stridulatory pattern generation in the grasshopper Chorthippus mollis (Charp.)

In tethered grasshoppers, Chorthippus mollis, stridulatory leg movements were elicited by d.c. brain stimulation. Stridulatory chirps comprise both slow up-and-down movements and rapid oscillations of the hindlegs. Intracellular recording, stimulation and staining of interneurones within the metathoracic ganglion complex were performed simultaneously with recordings of leg movement. Five interneurones were identified in the metathoracic ganglion complex. The branching patterns of these interneurones were typical of stridulatory interneurones. Three of these neurones had a structure very similar to stridulatory interneurones already characterized in the species Omocestus viridulus. During stridulation, the spike activity of all interneurones was phasically coupled to the chirp rhythm; two interneurones additionally exhibited coupling to the rapid leg oscillations. Intracellular stimulation of interneurones A1-AC-2 and A1-AI-1 prolonged the duration of the rapid leg oscillations and influenced the generation of the chirp rhythm. Interneurones T3-LI-2 and T3-LC-4 decreased the amplitude of the slow up-and-down movement. The data indicate that at least part of the metathoracic stridulatory network of C. mollis is organized in a structurally and functionally similar way to that of O. viridulus.

Journal Article↗
PubMed1988

Stratum lacunosum-moleculare interneurons of hippocampal CA1 region. II. Intrasomatic and intradendritic recordings of local circuit synaptic interactions.

Simultaneous intracellular recordings were obtained from stratum lacunosum-moleculare (L-M) interneurons and CA1 cells, and their local circuit synaptic interactions were examined. Synaptic interactions with pyramidal cells were evaluated in both intrasomatic and intradendritic pyramidal cell recordings. Stimulation of L-M interneurons evoked small-amplitude IPSPs in 21% of intrasomatic (9/42 cell pairs) and in 26% of intradendritic (11/43) pyramidal cell recordings. The IPSP mean peak amplitude was 0.91 mV for intrasomatic and 0.67 mV for intradendritic recordings. IPSPs had slow onset and decay (approximately 80-90 msec), decreased in amplitude with membrane hyperpolarization, and were not associated with any apparent change in input resistance. No physiologic evidence of synaptic connections was found from pyramidal cells to L-M interneurons. Inhibitory synaptic interactions were also seen between L-M interneurons and stratum pyramidale interneurons (2 of 4 cell pairs). The IPSPs recorded in pyramidale interneurons were similar to the IPSPs recorded in pyramidal cells. During simultaneous recordings, L-M interneurons were activated at a shorter latency, i.e., in a feedforward manner with respect to pyramidal cells. Thus, L-M interneurons may mediate feedforward inhibition of CA1 pyramidal cells. The L-M interneuron-evoked IPSPs in pyramidal cells share some characteristics of the late IPSP recorded in CA1 pyramidal cells and may therefore contribute to this component of the IPSP.

Animals↗
PubMed2001

Effects of serotonin depletion on local interneurons in the developing olfactory pathway of lobsters.

During embryonic life, the growth of the olfactory and accessory lobes of the lobster brain is retarded by serotonin depletion using 5,7-dihydroxytryptamine (5,7-DHT) (Benton et al., 1997). The local and projection interneurons that synapse with chemosensory cells in the olfactory lobes are potential targets of this depletion. This study documents proliferation and survival in the local interneuron cell clusters, and examines the differentiation of a prominent local interneuron, the serotonergic dorsal giant neuron (DGN), following serotonin depletion. An increase in dye coupling between the DGN and nearby cells is seen after serotonin depletion. However, morphometric analyses of individual DGNs in normal, sham-injected, and 5,7-DHT-treated embryos show that the general morphology and size of the DGNs are not significantly altered by serotonin depletion. Thus, the DGN axonal arbor occupies a greater proportion of the reduced olfactory lobes in the 5,7-DHT-treated embryos than in normal and sham-injected groups. The paired olfactory globular tract neutrophils (OGTNs), where olfactory interneurons synapse onto the DGNs, are 75% smaller in volume than the comparable region in either sham-injected or normal embryos. In vivo experiments using bromodeoxyuridine (BrdU) show that proliferation in the local interneuron soma clusters is reduced by 5,7-DHT treatment and that survival of newly proliferated local interneurons is also compromised. Our data suggest that alterations in the growth of the DGNs do not contribute to the dramatic reduction in size of the olfactory neutrophils following serotonin depletion, but that cell proliferation and survival among the local interneurons are regulated by serotonin during development. Reduced numbers of local interneurons are therefore one likely reason for the growth reduction observed after serotonin depletion.

5,7-Dihydroxytryptamine↗
PubMed2003

Loss of interneurons innervating pyramidal cell dendrites and axon initial segments in the CA1 region of the hippocampus following pilocarpine-induced seizures.

In the pilocarpine model of chronic limbic seizures, vulnerability of GABAergic interneurons to excitotoxic damage has been reported in the hippocampal CA1 region. However, little is known about the specific types of interneurons that degenerate in this region. In order to characterize these interneurons, we performed quantitative analyses of the different populations of GABAergic neurons labeled for their peptide or calcium-binding protein content. Our data demonstrate that the decrease in the number of GAD mRNA-containing neurons in the stratum oriens of CA1 in pilocarpine-treated rats involved two subpopulations of GABAergic interneurons: interneurons labeled for somatostatin only (O-LM and bistratified cells) and interneurons labeled for parvalbumin only (basket and axo-axonic cells). Stratum oriens interneurons labeled for somatostatin/calbindin or somatostatin/parvalbumin were preserved. The decrease in number of somatostatin- and parvalbumin-containing neurons was observed as early as 72 hours after the sustained seizures induced by pilocarpine injection. Many degenerating cell bodies in the stratum oriens and degenerating axon terminals in the stratum lacunosum-moleculare were observed at 1 and 2 weeks after injection. In addition, the synaptic coverage of the axon initial segment of CA1 pyramidal cells was significantly decreased in pilocarpine-treated animals. These results indicate that the loss of somatostatin-containing neurons corresponds preferentially to the degeneration of interneurons with an axon projecting to stratum lacunosum-moleculare (O-LM cells) and suggest that the death of these neurons is mainly responsible for the deficit of dendritic inhibition reported in this region. We demonstrate that the loss of parvalbumin-containing neurons corresponds to the death of axo-axonic cells, suggesting that perisomatic inhibition and mechanisms controlling action potential generation are also impaired in this model.

Animals↗
PubMed2003

Synaptic targets of commissural interneurons in the lumbar spinal cord of neonatal rats.

There is strong evidence that commissural interneurons, neurons with axons that extend to the contralateral side of the spinal cord, play an important role in the coordination of left/right alternation during locomotion. In this study we investigated the projections of commissural interneurons to motor neurons and other commissural interneurons on the other side of the spinal cord in neonatal rats. To establish whether there are direct contacts between axons of commissural interneurons and motor neurons, we carried out two series of experiments. In the first experiment we injected biotinylated dextran amine (BDA) into the lateral motor column to retrogradely label commissural interneurons that may have direct projections to motor neurons. Stained neurons were recovered in the ventromedial areas of the contralateral gray matter in substantial numbers. In the second experiment BDA was injected into the ventromedial gray matter on one side of the lumbar spinal cord, whereas motor neurons were simultaneously labeled on the opposite side by applying biocytin onto the ventral roots. BDA injections into the ventromedial gray matter labeled a strong axon bundle that arose from the site of injection, crossed the midline in the ventral commissure, and extensively arborized in the contralateral ventral gray matter. Many of these axons made close appositions with dendrites and somata of motor neurons and also with commissural interneurons retrogradely labeled with BDA. The results suggest that commissural interneurons may establish monosynaptic contacts with motor neurons on the opposite side of the spinal cord. Our findings also indicate that direct reciprocal connections between commissural interneurons on the two sides of the spinal cord may also exist.

Afferent Pathways↗