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1.
Neurochem Res ; 46(7): 1659-1673, 2021 Jul.
Article in English | MEDLINE | ID: mdl-33770320

ABSTRACT

Parvalbumin-immunoreactive (Parv+) interneurons is an important component of striatal GABAergic microcircuits, which receive excitatory inputs from the cortex and thalamus, and then target striatal projection neurons. The present study aimed to examine ultrastructural synaptic connection features of Parv+ neruons with cortical and thalamic input, and striatal projection neurons by using immuno-electron microscopy (immuno-EM) and immunofluorescence techniques. Our results showed that both Parv+ somas and dendrites received numerous asymmetric synaptic inputs, and Parv+ terminals formed symmetric synapses with Parv- somas, dendrites and spine bases. Most interestingly, spine bases targeted by Parv+ terminals simultaneously received excitatory inputs at their heads. Electrical stimulation of the motor cortex (M1) induced higher proportion of striatal Parv+ neurons express c-Jun than stimulation of the parafascicular nucleus (PFN), and indicated that cortical- and thalamic-inputs differentially modulate Parv+ neurons. Consistent with that, both Parv + soma and dendrites received more VGlut1+ than VGlut2+ terminals. However, the proportion of VGlut1+ terminal targeting onto Parv+ proximal and distal dendrites was not different, but VGlut2+ terminals tended to target Parv+ somas and proximal dendrites than distal dendrites. These functional and morphological results suggested excitatory cortical and thalamic glutamatergic inputs differently modulate Parv+ interneurons, which provided inhibition inputs onto striatal projection neurons. To maintain the balance between the cortex and thalamus onto Parv+ interneurons may be an important therapeutic target for neurological disorders.


Subject(s)
Cerebral Cortex/ultrastructure , Dendrites/ultrastructure , Interneurons/ultrastructure , Intralaminar Thalamic Nuclei/ultrastructure , Parvalbumins/metabolism , Synapses/ultrastructure , Animals , Cerebral Cortex/metabolism , Dendrites/metabolism , Interneurons/metabolism , Intralaminar Thalamic Nuclei/metabolism , Male , Rats, Sprague-Dawley , Synapses/metabolism , Vesicular Glutamate Transport Protein 1/metabolism , Vesicular Glutamate Transport Protein 2/metabolism
2.
Brain Struct Funct ; 224(9): 3321-3338, 2019 Dec.
Article in English | MEDLINE | ID: mdl-31679085

ABSTRACT

In both Parkinson's disease (PD) patients and MPTP-treated non-human primates, there is a profound neuronal degeneration of the intralaminar centromedian/parafascicular (CM/Pf) thalamic complex. Although this thalamic pathology has long been established in PD (and other neurodegenerative disorders), the impact of CM/Pf cell loss on the integrity of the thalamo-striatal glutamatergic system and its regulatory functions upon striatal neurons remain unknown. In the striatum, cholinergic interneurons (ChIs) are important constituents of the striatal microcircuitry and represent one of the main targets of CM/Pf-striatal projections. Using light and electron microscopy approaches, we have analyzed the potential impact of CM/Pf neuronal loss on the anatomy of the synaptic connections between thalamic terminals (vGluT2-positive) and ChIs neurons in the striatum of parkinsonian monkeys treated chronically with MPTP. The following conclusions can be drawn from our observations: (1) as reported in PD patients, and in our previous monkey study, CM/Pf neurons undergo profound degeneration in monkeys chronically treated with low doses of MPTP. (2) In the caudate (head and body) nucleus of parkinsonian monkeys, there is an increased density of ChIs. (3) Despite the robust loss of CM/Pf neurons, no significant change was found in the density of thalamostriatal (vGluT2-positive) terminals, and in the prevalence of vGluT2-positive terminals in contact with ChIs in parkinsonian monkeys. These findings provide new information about the state of thalamic innervation of the striatum in parkinsonian monkeys with CM/Pf degeneration, and bring up an additional level of intricacy to the consequences of thalamic pathology upon the functional microcircuitry of the thalamostriatal system in parkinsonism. Future studies are needed to assess the importance of CM/Pf neuronal loss, and its potential consequences on the neuroplastic changes induced in the synaptic organization of the thalamostriatal system, in the development of early cognitive impairments in PD.


Subject(s)
Caudate Nucleus/pathology , Cholinergic Neurons/pathology , Glutamic Acid , Intralaminar Thalamic Nuclei/pathology , Neurons/pathology , Parkinsonian Disorders/pathology , Putamen/pathology , Animals , Caudate Nucleus/ultrastructure , Cholinergic Neurons/ultrastructure , Female , Interneurons/pathology , Interneurons/ultrastructure , Intralaminar Thalamic Nuclei/ultrastructure , Macaca mulatta , Male , Neural Pathways/pathology , Neural Pathways/ultrastructure , Neurons/ultrastructure , Putamen/ultrastructure , Synapses/pathology , Synapses/ultrastructure , Vesicular Glutamate Transport Protein 2/metabolism
3.
Brain Struct Funct ; 224(8): 2787-2804, 2019 Nov.
Article in English | MEDLINE | ID: mdl-31422483

ABSTRACT

Preclinical evidence indicates that mGluR5 is a potential therapeutic target for Parkinson's disease and L-DOPA-induced dyskinesia. However, the mechanisms through which these therapeutic benefits are mediated remain poorly understood. Although the regulatory role of mGluR5 on glutamatergic transmission has been examined in various basal ganglia nuclei, very little is known about the localization and function of mGluR5 in the ventral motor and intralaminar thalamic nuclei, the main targets of basal ganglia output in mammals. Thus, we used immuno-electron microscopy to map the cellular and subcellular localization of group I mGluRs (mGluR1a and mGluR5) in the ventral motor and caudal intralaminar thalamic nuclei in rhesus monkeys. Furthermore, using double immuno-electron microscopy, we examined the subsynaptic localization of mGluR5 in relation to cortical and sub-cortical glutamatergic afferents. Four major conclusions can be drawn from these data. First, mGluR1a and mGluR5 are expressed postsynaptically on the plasma membrane of dendrites of projection neurons and GABAergic interneurons in the basal ganglia- and cerebellar-receiving regions of the ventral motor thalamus and in CM. Second, the plasma membrane-bound mGluR5 immunoreactivity is preferentially expressed perisynaptically at the edges of cortical and sub-cortical glutamatergic afferents. Third, the mGluR5 immunoreactivity is more strongly expressed in the lateral than the medial tiers of CM, suggesting a preferential association with thalamocortical over thalamostriatal neurons in the primate CM. Overall, mGluR5 is located to subserve powerful modulatory role of cortical and subcortical glutamatergic transmission in the primate ventral motor thalamus and CM.


Subject(s)
Cerebral Cortex/ultrastructure , Neurons/ultrastructure , Presynaptic Terminals/ultrastructure , Receptor, Metabotropic Glutamate 5/analysis , Receptors, Metabotropic Glutamate/analysis , Thalamus/ultrastructure , Afferent Pathways/ultrastructure , Animals , Dendrites/ultrastructure , Female , Intralaminar Thalamic Nuclei/ultrastructure , Macaca mulatta , Male
4.
Brain Struct Funct ; 222(2): 735-748, 2017 03.
Article in English | MEDLINE | ID: mdl-27255751

ABSTRACT

T-type calcium channels (Cav3) are key mediators of thalamic bursting activity, but also regulate single cells excitability, dendritic integration, synaptic strength and transmitter release. These functions are strongly influenced by the subcellular and subsynaptic localization of Cav3 channels along the somatodendritic domain of thalamic cells. In Parkinson's disease, T-type calcium channels dysfunction in the basal ganglia-receiving thalamic nuclei likely contributes to pathological thalamic bursting activity. In this study, we analyzed the cellular, subcellular, and subsynaptic localization of the Cav3.1 channel in the ventral anterior (VA) and centromedian/parafascicular (CM/Pf) thalamic nuclei, the main thalamic targets of basal ganglia output, in normal and parkinsonian monkeys. All thalamic nuclei displayed strong Cav3.1 neuropil immunoreactivity, although the intensity of immunolabeling in CM/Pf was significantly lower than in VA. Ultrastructurally, 70-80 % of the Cav3.1-immunoreactive structures were dendritic shafts. Using immunogold labeling, Cav3.1 was commonly found perisynaptic to asymmetric and symmetric axo-dendritic synapses, suggesting a role of Cav3.1 in regulating excitatory and inhibitory neurotransmission. Significant labeling was also found at non-synaptic sites along the plasma membrane of thalamic neurons. There was no difference in the overall pattern and intensity of immunostaining between normal and parkinsonian monkeys, suggesting that the increased rebound bursting in the parkinsonian state is not driven by changes in Cav3.1 expression. Thus, T-type calcium channels are located to subserve neuronal bursting, but also regulate glutamatergic and non-glutamatergic transmission along the whole somatodendritic domain of basal ganglia-receiving neurons of the primate thalamus.


Subject(s)
Calcium Channels, T-Type/metabolism , Neurons/metabolism , Parkinson Disease/metabolism , Synapses/metabolism , Thalamus/metabolism , Animals , Disease Models, Animal , Female , Intralaminar Thalamic Nuclei/metabolism , Intralaminar Thalamic Nuclei/ultrastructure , Macaca mulatta , Neurons/ultrastructure , Parkinsonian Disorders/metabolism , Synapses/ultrastructure , Thalamus/ultrastructure , Ventral Thalamic Nuclei/metabolism , Ventral Thalamic Nuclei/ultrastructure
5.
J Neurosci Res ; 88(2): 448-60, 2010 Feb 01.
Article in English | MEDLINE | ID: mdl-19774668

ABSTRACT

Low-voltage-activated (LVA) Ca(2+) channels play a critical role in the generation of burst firing in the thalamus. Recently, three LVA Ca(2+) channel isoforms (Ca(v)3.1, Ca(v)3.2, Ca(v)3.3) have been identified in the reticular thalamic nucleus (RE). Previous electrophysiological and modelling studies have suggested that kinetically different T-type channels might be expressed in a compartmentalized manner in RE cells. However, their precise subcellular distribution has not been fully elucidated. Using light and electron microscopic (EM) immunocytochemistry, we investigated the subcellular expression pattern of Ca(v)3.1 and Ca(v)3.3 channel subunits in RE neurons of the cat. Fluorescent and peroxidase labelling demonstrated the presence of Ca(v)3.1 channel predominantly on the somata and proximal dendrites and Ca(v)3.3 channels on cell bodies. Quantitative immunogold localization disclosed that Ca(v)3.1 and Ca(v)3.3 isoforms showed 5.8- and 8.7-fold higher density, respectively, in the cytoplasm compared with somatic plasma membrane. Density of Ca(v)3.1 isoform in the somatic plasma membrane was 2.21-fold higher compared with Ca(v)3.3 subunit. In the dendritic plasma membrane, Ca(v)3.1 channel isoform was expressed throughout the entire dendritic tree. In contrast, Ca(v)3.3 isoform was absent from large-caliber, presumably proximal dendritic segments. Quantitative comparison showed that the relative density of immunogold particles compared with dendritic surface was 8.9- and 14.8-fold higher for Ca(v)3.1 and Ca(v)3.3, respectively, in small-diameter dendrites than in large proximal dendritic segments or somata. Our results demonstrate a higher density of low-threshold Ca(2+) channels in distal dendrites and provide further evidence of the role of RE neuron dendrites in the generation of prolonged, low-threshold spike bursts.


Subject(s)
Calcium Channels, T-Type/metabolism , Intralaminar Thalamic Nuclei/metabolism , Neurons/metabolism , Animals , Blotting, Western , Cats , Cell Membrane/metabolism , Cell Membrane/ultrastructure , Cytoplasm/metabolism , Cytoplasm/ultrastructure , Dendrites/metabolism , Dendrites/ultrastructure , Fluorescence , Immunoenzyme Techniques , Immunohistochemistry , Intralaminar Thalamic Nuclei/ultrastructure , Microscopy, Electron , Neurons/ultrastructure , Parvalbumins/metabolism , Protein Isoforms/metabolism , Subcellular Fractions/metabolism , Subcellular Fractions/ultrastructure , Thalamic Nuclei/metabolism , Thalamic Nuclei/ultrastructure
6.
J Chem Neuroanat ; 37(1): 33-45, 2009 Jan.
Article in English | MEDLINE | ID: mdl-18773952

ABSTRACT

Cholinergic interneurons are the only known source of acetylcholine in the rat nucleus accumbens (nAcb); yet there is little anatomical data about their mode of innervation and the origin of their excitatory drive. We characterized the cholinergic and thalamic innervations of nAcb with choline acetyltransferase (ChAT) immunocytochemistry and anterograde transport of Phaseolus vulgaris-leucoagglutinin (PHA-L) from the midline/intralaminar/paraventricular thalamic nuclei. The use of a monoclonal ChAT antiserum against whole rat ChAT protein allowed for an optimal visualization of the small dendritic branches and fine varicose axons of cholinergic interneurons. PHA-L-labeled thalamic afferents were heterogeneously distributed throughout the core and shell regions of nAcb, overlapping regionally with cholinergic somata and dendrites. At the ultrastructural level, several hundred single-section profiles of PHA-L and ChAT-labeled axon terminals were analyzed for morphology, synaptic frequency, and the nature of their synaptic targets. The cholinergic profiles were small and apposed to various neuronal elements, but rarely exhibited a synaptic membrane specialization (5% in single ultrathin sections). Stereological extrapolation indicated that less than 15% of these cholinergic varicosities were synaptic. The PHA-L-labeled profiles were comparatively large and often synaptic (37% in single ultrathin sections), making asymmetrical contacts primarily with dendritic spines (>90%). Stereological extrapolation indicated that all PHA-L-labeled terminals were synaptic. In double-labeled material, some PHA-L-labeled terminals were directly apposed to ChAT-labeled somata or dendrites, but synapses were never seen between the two types of elements. These observations demonstrate that the cholinergic innervation of rat nAcb is largely asynaptic. They confirm that the afferents from midline/intralaminar/paraventricular thalamic nuclei to rat nAcb synapse mostly on dendritic spines, presumably of medium spiny neurons, and suggest that the excitatory drive of nAcb cholinergic interneurons from thalamus is indirect, either via substance P release from recurrent collaterals of medium spiny neurons and/or by extrasynaptic diffusion of glutamate.


Subject(s)
Choline O-Acetyltransferase/physiology , Nucleus Accumbens/physiology , Thalamus/physiology , Afferent Pathways/physiology , Animals , Antibodies, Monoclonal , Female , Immunohistochemistry , Interneurons/physiology , Intralaminar Thalamic Nuclei/physiology , Intralaminar Thalamic Nuclei/ultrastructure , Male , Midline Thalamic Nuclei/physiology , Midline Thalamic Nuclei/ultrastructure , Nucleus Accumbens/ultrastructure , Phaseolus , Phytohemagglutinins , Rats , Rats, Wistar , Synapses/physiology
7.
J Neurosci ; 28(46): 11848-61, 2008 Nov 12.
Article in English | MEDLINE | ID: mdl-19005050

ABSTRACT

Diverse sources of GABAergic inhibition are a major feature of cortical networks, but distinct inhibitory input systems have not been systematically characterized in the thalamus. Here, we contrasted the properties of two independent GABAergic pathways in the posterior thalamic nucleus of rat, one input from the reticular thalamic nucleus (nRT), and one "extrareticular" input from the anterior pretectal nucleus (APT). The vast majority of nRT-thalamic terminals formed single synapses per postsynaptic target and innervated thin distal dendrites of relay cells. In contrast, single APT-thalamic terminals formed synaptic contacts exclusively via multiple, closely spaced synapses on thick relay cell dendrites. Quantal analysis demonstrated that the two inputs displayed comparable quantal amplitudes, release probabilities, and multiple release sites. The morphological and physiological data together indicated multiple, single-site contacts for nRT and multisite contacts for APT axons. The contrasting synaptic arrangements of the two pathways were paralleled by different short-term plasticities. The multisite APT-thalamic pathway showed larger charge transfer during 50-100 Hz stimulation compared with the nRT pathway and a greater persistent inhibition accruing during stimulation trains. Our results demonstrate that the two inhibitory systems are morpho-functionally distinct and suggest and that multisite GABAergic terminals are tailored for maintained synaptic inhibition even at high presynaptic firing rates. These data explain the efficacy of extrareticular inhibition in timing relay cell activity in sensory and motor thalamic nuclei. Finally, based on the classic nomenclature and the difference between reticular and extrareticular terminals, we define a novel, multisite GABAergic terminal type (F3) in the thalamus.


Subject(s)
Intralaminar Thalamic Nuclei/metabolism , Posterior Thalamic Nuclei/metabolism , Presynaptic Terminals/metabolism , Synapses/metabolism , gamma-Aminobutyric Acid/metabolism , Afferent Pathways/metabolism , Afferent Pathways/ultrastructure , Animals , Dendrites/metabolism , Dendrites/ultrastructure , Electric Stimulation , Inhibitory Postsynaptic Potentials/physiology , Intralaminar Thalamic Nuclei/ultrastructure , Male , Microscopy, Immunoelectron , Neural Inhibition/physiology , Posterior Thalamic Nuclei/ultrastructure , Presynaptic Terminals/ultrastructure , Rats , Rats, Wistar , Superior Colliculi/metabolism , Superior Colliculi/ultrastructure , Synaptic Transmission/physiology
8.
J Comp Neurol ; 511(5): 678-91, 2008 Dec 10.
Article in English | MEDLINE | ID: mdl-18924144

ABSTRACT

The acetylcholine (ACh) innervation of thalamus arises mainly from the brainstem pedunculopontine and laterodorsal tegmental nuclei. By using immunocytochemistry with a monoclonal antibody against whole rat choline acetyltransferase (ChAT), we quantified the distribution and characterized the ultrastructural features of these nerve terminals (axon varicosities) in the dorsolateral geniculate (DLG), parafascicular (PF), and reticular thalamic (Rt) nuclei of adult rat. The regional density of ACh innervation was the highest in PF (2.1 x 10(6) varicosities/mm(3)), followed by Rt (1.7 x 10(6)) and DLG (1.3 x 10(6)). In single thin sections, ChAT-immunostained varicosity profiles appeared comparable in shape and content in the three nuclei, but significantly larger in PF than in DLG and Rt. The number of these profiles displaying a synaptic junction was also much higher in PF than in DLG and Rt, indicating that all ChAT-immunostained varicosities in PF were synaptic, but only 39% in DLG and 33% in Rt. The hypothesis that glutamate corelease might account for the maintenance of the entirely synaptic ACh innervation in PF was refuted by the lack of colocalization of ChAT and vesicular glutamate transporter 2 (VGLUT2) in PF axon varicosities after dual immunolabeling. These data suggest that diffuse as well as synaptic transmission convey modulatory effects of the ACh input from brainstem to DLG and Rt during waking. In contrast, the entirely synaptic ACh input to PF should allow for a direct relaying of the information from brainstem, affecting basal ganglia function as well as perceptual awareness, including attention and pain perception.


Subject(s)
Acetylcholine/metabolism , Cholinergic Fibers/metabolism , Pedunculopontine Tegmental Nucleus/metabolism , Presynaptic Terminals/metabolism , Thalamus/metabolism , Animals , Arousal/physiology , Choline O-Acetyltransferase/metabolism , Cholinergic Fibers/ultrastructure , Geniculate Bodies/metabolism , Geniculate Bodies/ultrastructure , Glutamic Acid/metabolism , Intralaminar Thalamic Nuclei/metabolism , Intralaminar Thalamic Nuclei/ultrastructure , Male , Microscopy, Immunoelectron , Neural Pathways/metabolism , Neural Pathways/ultrastructure , Pedunculopontine Tegmental Nucleus/ultrastructure , Presynaptic Terminals/ultrastructure , Rats , Rats, Sprague-Dawley , Synaptic Transmission/physiology , Thalamus/ultrastructure , Vesicular Glutamate Transport Protein 2/metabolism , Wakefulness/physiology
9.
J Comp Neurol ; 504(5): 583-98, 2007 Oct 10.
Article in English | MEDLINE | ID: mdl-17701985

ABSTRACT

The lateral reticular nucleus (LRN) resides in the rostral medulla and caudal pons, is implicated in cardiovascular regulation and cranial nerve reflexes, and gives rise to mossy fibers in the cerebellum. Retrograde tracing data revealed that medium-sized multipolar cells from the magnocellular part of the LRN project to the cochlear nucleus (CN). We sought to characterize the LRN projection to the CN using BDA injections. Anterogradely labeled terminals in the ipsilateral CN appeared as boutons and mossy fibers, and were examined with light and electron microscopy. The terminal field in the CN was restricted to the granule cell domain (GCD), specifically in the superficial layer along the anteroventral CN and in the granule cell lamina. Electron microscopy showed that the smallest LRN boutons formed 1-3 synapses, and as boutons increased in size, they formed correspondingly more synapses. The largest boutons were indistinguishable from the smallest mossy fibers, and the largest mossy fiber exhibited 15 synapses. Synapses were asymmetric with round vesicles and formed against thin dendritic profiles characterized by plentiful microtubules and the presence of fine filopodial extensions that penetrated the ending. These structural features of the postsynaptic target are characteristic of the terminal dendritic claw of granule cells. LRN projections are consistent with known organizational principles of non-auditory inputs to the GCD.


Subject(s)
Cochlear Nucleus/physiology , Intralaminar Thalamic Nuclei/physiology , Amidines/metabolism , Animals , Biotin/analogs & derivatives , Biotin/metabolism , Cochlear Nucleus/ultrastructure , Dextrans/metabolism , Intralaminar Thalamic Nuclei/ultrastructure , Microscopy, Electron, Transmission/trends , Nerve Fibers/ultrastructure , Neural Pathways/physiology , Presynaptic Terminals/ultrastructure , Rats
10.
Neurosci Res ; 56(1): 73-9, 2006 Sep.
Article in English | MEDLINE | ID: mdl-16814420

ABSTRACT

A disynaptic projection from the spinal cord to the striatum was observed in the rat light and electron microscopically. An anterograde tracer, wheat germ agglutinin conjugated to horseradish peroxidase was injected into the ventral gray matter of the upper cervical spinal cord, and a retrograde tracer, biotinylated dextran amine was injected into the striatum of a rat. Then the parafascicular nucleus was examined. Some anterogradely labeled axon terminals originating in the spinal cord were observed to synapse with retrogradely labeled dendrites of parafascicular nucleus neurons which sent axons to the striatum. We concluded that information from the spinal cord was transmitted to the striatum, being relayed by parafascicular nucleus neurons.


Subject(s)
Afferent Pathways/ultrastructure , Corpus Striatum/cytology , Intralaminar Thalamic Nuclei/ultrastructure , Spinal Cord/cytology , Animals , Male , Neurons/ultrastructure , Rats , Rats, Wistar , Staining and Labeling
11.
Morfologiia ; 128(6): 9-17, 2005.
Article in Russian | MEDLINE | ID: mdl-16755783

ABSTRACT

This review presents an analysis of current data on the structural organization of the reticular thalamic nucleus (RTN). In contains a detailed discussion on topography, cytoarchitecture, neuronal organization and neurogenesis in this nucleus. The materials on RTN neurochemistry are systematized specifying the topographic representation of neurotransmitter systems and neuropeptides which are synthesized in RTN neurons. Complex ultrastructural organization of RTN is characterized on the basis of modern immunocytochemical methods, which allow to detect the localization of glutamatergic and GABAergic receptors on synaptic elements. Data on afferent and efferent connections of RTN demonstrate its influence on various brain regions and the specificity of RTN interrelations with some cortical formations.


Subject(s)
Intralaminar Thalamic Nuclei/ultrastructure , Animals , Humans , Immunohistochemistry , Intralaminar Thalamic Nuclei/chemistry , Neurons/chemistry , Neurons/ultrastructure , Receptors, Amino Acid/analysis
12.
Eur J Neurosci ; 20(4): 965-75, 2004 Aug.
Article in English | MEDLINE | ID: mdl-15305865

ABSTRACT

In the thalamus of the rat the reversal potential of GABA-induced anion currents is more negative in relay cells than in neurones of the reticular nucleus (nRt) due to different chloride extrusion mechanisms operating in these cells. The distribution of KCl cotransporter type 2 (KCC2), the major neuronal chloride transporter that may underlie this effect, is unknown in the thalamus. In this study the precise regional and ultrastructural localization of KCC2 was examined in the thalamus using immunocytochemical methods. The neuropil of all relay nuclei was found to display intense KCC2 immunostaining to varying degrees. In sharp contrast, the majority of the nRt was negative for KCC2. In the anterior and dorsal part of the nRt, however, KCC2 immunostaining was similar to relay nuclei and parvalbumin and calretinin were found to colocalize with KCC2. At the ultrastructural level, KCC2 immunoreactivity was mainly located in the extrasynaptic membranes of thick and thin dendrites and the somata of relay cells but was also found in close association with asymmetrical synapses formed by cortical afferents. Quantitative evaluation of KCC2 distribution at the electron microscopic level demonstrated that the density of KCC2 did not correlate with dendritic diameter or synaptic coverage but is 1.7 times higher on perisynaptic membrane surfaces than on extrasynaptic membranes. Our data demonstrate that the regional distribution of KCC2 is compatible with the difference in GABA-A reversal potential between relay and reticular nuclei. At the ultrastructural level, abundant extrasynaptic KCC2 expression will probably play a role in the regulation of extrasynaptic GABA-A receptor-mediated inhibition.


Subject(s)
Symporters/metabolism , Thalamic Nuclei/chemistry , Thalamic Nuclei/metabolism , Animals , Dendrites/metabolism , Dendrites/physiology , Dendrites/ultrastructure , Intralaminar Thalamic Nuclei/chemistry , Intralaminar Thalamic Nuclei/metabolism , Intralaminar Thalamic Nuclei/ultrastructure , Male , Neural Inhibition/physiology , Rats , Rats, Wistar , Subcellular Fractions/metabolism , Subcellular Fractions/physiology , Subcellular Fractions/ultrastructure , Synaptic Membranes/metabolism , Synaptic Membranes/physiology , Synaptic Membranes/ultrastructure , Thalamic Nuclei/ultrastructure , K Cl- Cotransporters
13.
Brain Res ; 957(2): 231-41, 2002 Dec 13.
Article in English | MEDLINE | ID: mdl-12445965

ABSTRACT

The ventrolateral part of the parafascicular thalamic nucleus (PF), which is considered to take part in the control mechanism of orofacial motor functions, receives projection fibers not only from the dorsolateral part of the substantia nigra pars reticulata (SNr) but also from the ventral part of the reticular thalamic nucleus (RT) [Tsumori et al., Brain Res. 858 (2000) 429]. In order to better understand the influence of these fibers upon the PF projection neurons, the morphology, synaptology and chemical nature of them were examined in the present study. After ipsilateral injections of Phaseolus vulgaris-leucoagglutinin (PHA-L) into the dorsolateral part of the SNr and biotinylated dextran amine (BDA) into the ventral part of the RT, overlapping distributions of PHA-L-labeled SNr fibers and BDA-labeled RT fibers were seen in the ventrolateral part of the PF. At the electron microscopic level, the SNr terminals made synapses predominantly with the medium to small dendrites and far less frequently with the somata and large dendrites, whereas approximately half of the RT terminals made synapses with the somata and large dendrites and the rest did with the medium to small dendrites of PF neurons. Some of single dendritic as well as single somatic profiles received convergent synaptic inputs from both sets of terminals. These terminals were packed with pleomorphic synaptic vesicles and formed symmetrical synapses. After combined injections of PHA-L into the dorsolateral part of the SNr, BDA into the ventral part of the RT and wheat germ agglutinin-horseradish peroxidase (WGA-HRP) into the ventrolateral part of the striatum or into the rostroventral part of the lateral agranular cortex, WGA-HRP-labeled neurons were embedded in the plexus of PHA-L- and BDA-labeled axon terminals within the ventrolateral part of the PF, where the PHA-L- and/or BDA-labeled terminals were in synaptic contact with single somatic and dendritic profiles of the WGA-HRP-labeled neurons. Furthermore, the SNr and RT axon terminals were revealed to be immunoreactive for gamma-aminobutyric acid (GABA), by using the anterograde BDA tracing technique combined with immunohistochemistry for GABA. The present data suggest that GABAergic SNr and RT fibers may exert different inhibitory influences on the PF neurons for regulating the thalamic outflow from the PF to the cerebral cortex and/or striatum in the control of orofacial movements.


Subject(s)
Intralaminar Thalamic Nuclei/ultrastructure , Neural Inhibition/physiology , Neural Pathways/ultrastructure , Presynaptic Terminals/ultrastructure , Substantia Nigra/ultrastructure , gamma-Aminobutyric Acid/metabolism , Animals , Corpus Striatum/metabolism , Corpus Striatum/ultrastructure , Dendrites/metabolism , Dendrites/ultrastructure , Immunohistochemistry , Intralaminar Thalamic Nuclei/metabolism , Male , Microscopy, Electron , Motor Cortex/metabolism , Motor Cortex/ultrastructure , Neural Pathways/metabolism , Presynaptic Terminals/metabolism , Rats , Rats, Wistar , Substantia Nigra/metabolism , Synaptic Transmission/physiology
14.
Prog Brain Res ; 136: 333-57, 2002.
Article in English | MEDLINE | ID: mdl-12143393

ABSTRACT

Cajal's many contributions to understanding the thalamus have been hidden by his body of work on the cerebral cortex. He delineated many thalamic nuclei in rodents, defined afferent fibers, thalamocortical relay neurons and interneurons, was first to demonstrate thalamocortical fibers and their terminations in the cortex, and recognized the feed-back provided by corticothalamic fibers. This presentation outlines modern methods for identifying classes of thalamic neurons, their chemical characteristics, synaptology and differential connections, and describes the intrinsic circuitry of the thalamus, showing how interactions between GABAergic cells of the reticular nucleus and glutamatergic relay cells underlie rhythmic activities of neurons in the thalamo-cortico-thalamic network, activities associated with changes in the conscious state, and which are generated and maintained by the corticothalamic projection. Corticothalamic fibers interact with reticular nucleus cells and relay cells through NMDA, AMPA and metabotropic receptors while interactions between reticular nucleus cells and relay cells are mediated by GABAA and GABAB receptors. Differing strengths of synaptic input to the two cell types, from which oscillatory behavior commences, depend upon differential expression at individual synapses of specific AMPA receptor subunits which modulate excitatory postsynaptic conductances. Two classes of relay cells can be distinguished by differential staining for calbindin and parvalbumin. The first forms a matrix in the thalamus, unconstrained by nuclear borders; the second is concentrated in certain nuclei in which it forms the topographically organized core. In projecting diffusely to the cortex, calbindin cells provide a substrate for binding together activities of multiple cortical areas that receive focused input from single thalamic nuclei. This, and the presence of specific and diffuse corticothalamic projections may serve to promote coherent activity of large populations of cortical and thalamic neurons in perception, attention and conscious awareness.


Subject(s)
Neural Pathways/ultrastructure , Neurons/ultrastructure , Thalamus/ultrastructure , Animals , Biological Clocks/physiology , Cerebral Cortex/metabolism , Cerebral Cortex/ultrastructure , Humans , Interneurons/metabolism , Interneurons/ultrastructure , Intralaminar Thalamic Nuclei/metabolism , Intralaminar Thalamic Nuclei/ultrastructure , Neural Inhibition/physiology , Neural Pathways/metabolism , Neurons/metabolism , Presynaptic Terminals/metabolism , Presynaptic Terminals/ultrastructure , Thalamus/metabolism
15.
J Comp Neurol ; 447(3): 286-99, 2002 Jun 03.
Article in English | MEDLINE | ID: mdl-11984822

ABSTRACT

In primates, thalamostriatal projections from the centromedian (CM) and parafascicular (Pf) nuclei are strong and organized according to a strict pattern of functional connectivity with various regions of the striatal complex. In turn, the CM/Pf complex receives a substantial innervation from the internal globus pallidus (GPi). In this study, we demonstrate that the substantia nigra pars reticulata (SNr) also provides a massive input to Pf in monkeys. These pallidothalamic and nigrothalamic projections provide routes whereby information can flow in functional loops between the basal ganglia and the intralaminar nuclear group. To understand better the anatomical organization and the degree of functional specificity of these loops, we combined retrograde and anterograde labeling methods from functionally defined regions of the striatum and GPi/SNr to determine the relationships between thalamostriatal neurons and basal ganglia afferents. Together with previous studies, our data suggest the existence of tightly connected functional circuits between the basal ganglia and the CM/Pf in primates: 1) A "sensorimotor" circuit links together the medial two-thirds of CM, the postcommissural putamen, and the ventrolateral part of the caudal GPi; 2) a "limbic" circuit involves the rostral one-third of Pf, the ventral striatum, and the rostromedial pole of GPi; and 3) an "associative"circuit exists between the caudal two-thirds of Pf, the caudate nucleus, and the SNr. An additional "associative" circuit that involves the caudate-receiving territory of GPi (dorsal one-third), the dorsolateral Pf (Pfdl), and the precommissural putamen was also disclosed. In conclusion, findings of this study provide additional evidence for the high degree of functional specificity of the thalamostriatal system through which CM/Pf may provide attention-specific sensory information important for conditional responses to the primate striatum.


Subject(s)
Biotin/analogs & derivatives , Globus Pallidus/ultrastructure , Intralaminar Thalamic Nuclei/ultrastructure , Neostriatum/ultrastructure , Neural Pathways/ultrastructure , Presynaptic Terminals/ultrastructure , Saimiri/anatomy & histology , Substantia Nigra/ultrastructure , Animals , Caudate Nucleus/physiology , Caudate Nucleus/ultrastructure , Cholera Toxin/metabolism , Dextrans , Globus Pallidus/physiology , Intralaminar Thalamic Nuclei/physiology , Male , Microscopy, Electron , Neostriatum/physiology , Nerve Net/physiology , Nerve Net/ultrastructure , Neural Pathways/physiology , Presynaptic Terminals/physiology , Putamen/physiology , Putamen/ultrastructure , Saimiri/physiology , Substantia Nigra/physiology , Wheat Germ Agglutinin-Horseradish Peroxidase Conjugate
16.
Neurosci Lett ; 302(2-3): 105-8, 2001 Apr 20.
Article in English | MEDLINE | ID: mdl-11290398

ABSTRACT

We examined light and electron microscopically intrastriatal targets of projection fibers from the central lateral thalamic nucleus (CL), which is a major relay of cerebello-striatal projections. The study was done in the rat by combining the anterograde tract-tracing with immunohistochemistry for parvalbumin (PV); an anterograde tracer (biotin dextran amine: BDA) was injected into the CL. In the striatum, 91% of BDA-labeled axon terminals made asymmetrical synapses on PV immunonegative dendritic spines (assumed to be those of striatal projection neurons); only 0.5% of BDA-labeled axon terminals made synapses on PV immunopositive dendritic shafts. The remaining BDA-labeled axon terminals were in synaptic contact with PV immunonegative dendritic shafts. The results suggest that the cerebello-striatal projections through the CL predominantly access to striatal projection neurons, with only minor access to PV immunopositive (assumed to be GABAergic) interneurons in the striatum.


Subject(s)
Biotin/analogs & derivatives , Intralaminar Thalamic Nuclei/ultrastructure , Neostriatum/ultrastructure , Neural Pathways/ultrastructure , Presynaptic Terminals/ultrastructure , Animals , Biotin/pharmacokinetics , Dendrites/metabolism , Dendrites/ultrastructure , Dextrans/pharmacokinetics , Fluorescent Dyes/pharmacokinetics , Immunohistochemistry , Intralaminar Thalamic Nuclei/metabolism , Male , Microscopy, Electron , Neostriatum/metabolism , Neural Pathways/metabolism , Parvalbumins/metabolism , Presynaptic Terminals/metabolism , Rats , Rats, Wistar
17.
Neuroscience ; 101(4): 969-82, 2000.
Article in English | MEDLINE | ID: mdl-11113346

ABSTRACT

Although the mechanisms by which the cerebral cortex controls its ascending input are still poorly understood, it is known that cortical control at the thalamic level is via direct glutamatergic projections to relay nuclei and to the reticular nucleus. Here we confirm previous light microscopic reports of a high expression of the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor subunit, GluR4, in reticular and ventral posterior thalamic nuclei of the rat, and moderate staining using an antibody recognizing both GluR2 and GluR3. In contrast only low levels of staining for GluR2, and barely detectable levels of GluR1 immunoreactivity were observed. After injections of biotinylated dextran, electron microscopy revealed that anterogradely-labeled cortical synapses in both thalamic nuclei were small with fewer mitochondria and more densely-packed vesicles than terminals likely to arise from intrinsic and ascending pathways. We performed post-embedding immunogold to provide quantitative data on the density of AMPA receptor subunits at morphologically-defined groups of synapses. We found that corticothalamic synapses in the reticular thalamic nucleus contain twice as much GluR2/3, and at least three times more GluR4 protein than do intrathalamic synapses. In the ventral posterior nucleus, corticothalamic synapses contain similar amounts of GluR2/3, but four times more GluR4 than do those from ascending afferents. Corticothalamic synapses in reticular nucleus contain slightly more GluR2/3, and three times more GluR4, than those in ventral posterior nucleus. We conclude that enrichment of GluR4 at morphologically-defined cortical synapses is a feature common to both thalamic nuclei, and those in the reticular nucleus express higher levels of AMPA receptors. The rapid kinetics of GluR4-rich AMPA receptors we suggest indicate that cortical descending control may be more temporally precise than previously recognized.


Subject(s)
Intralaminar Thalamic Nuclei/metabolism , Receptors, AMPA/metabolism , Synapses/metabolism , Ventral Thalamic Nuclei/metabolism , Animals , Immunohistochemistry , Intralaminar Thalamic Nuclei/ultrastructure , Male , Microscopy, Electron , Protein Isoforms/metabolism , Rats , Rats, Sprague-Dawley , Tissue Distribution , Ventral Thalamic Nuclei/ultrastructure
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