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1.
Nature ; 537(7620): 427-431, 2016 09 15.
Article in English | MEDLINE | ID: mdl-27556946

ABSTRACT

Mice deficient in the DNA excision-repair gene Ercc1 (Ercc1∆/-) show numerous accelerated ageing features that limit their lifespan to 4-6 months. They also exhibit a 'survival response', which suppresses growth and enhances cellular maintenance. Such a response resembles the anti-ageing response induced by dietary restriction (also known as caloric restriction). Here we report that a dietary restriction of 30% tripled the median and maximal remaining lifespans of these progeroid mice, strongly retarding numerous aspects of accelerated ageing. Mice undergoing dietary restriction retained 50% more neurons and maintained full motor function far beyond the lifespan of mice fed ad libitum. Other DNA-repair-deficient, progeroid Xpg-/- (also known as Ercc5-/-) mice, a model of Cockayne syndrome, responded similarly. The dietary restriction response in Ercc1∆/- mice closely resembled the effects of dietary restriction in wild-type animals. Notably, liver tissue from Ercc1∆/- mice fed ad libitum showed preferential extinction of the expression of long genes, a phenomenon we also observed in several tissues ageing normally. This is consistent with the accumulation of stochastic, transcription-blocking lesions that affect long genes more than short ones. Dietary restriction largely prevented this declining transcriptional output and reduced the number of γH2AX DNA damage foci, indicating that dietary restriction preserves genome function by alleviating DNA damage. Our findings establish the Ercc1∆/- mouse as a powerful model organism for health-sustaining interventions, reveal potential for reducing endogenous DNA damage, facilitate a better understanding of the molecular mechanism of dietary restriction and suggest a role for counterintuitive dietary-restriction-like therapy for human progeroid genome instability syndromes and possibly neurodegeneration in general.


Subject(s)
Aging/genetics , Caloric Restriction , DNA Repair/genetics , Diet, Reducing , Genomic Instability , Animals , Brain/physiology , DNA Damage , DNA-Binding Proteins/deficiency , DNA-Binding Proteins/genetics , Endonucleases/deficiency , Endonucleases/genetics , Female , Male , Mice , Neurodegenerative Diseases/genetics , Neurodegenerative Diseases/prevention & control , Nuclear Proteins/deficiency , Nuclear Proteins/genetics , Transcription Factors/deficiency , Transcription Factors/genetics , Transcriptome
3.
Mol Psychiatry ; 21(3): 364-75, 2016 03.
Article in English | MEDLINE | ID: mdl-25802982

ABSTRACT

Memories are encoded within sparsely distributed neuronal ensembles. However, the defining cellular properties of neurons within a memory trace remain incompletely understood. Using a fluorescence-based Arc reporter, we were able to visually identify the distinct subset of lateral amygdala (LA) neurons activated during auditory fear conditioning. We found that Arc-expressing neurons have enhanced intrinsic excitability and are preferentially recruited into newly encoded memory traces. Furthermore, synaptic potentiation of thalamic inputs to the LA during fear conditioning is learning-specific, postsynaptically mediated and highly localized to Arc-expressing neurons. Taken together, our findings validate the immediate-early gene Arc as a molecular marker for the LA neuronal ensemble recruited during fear learning. Moreover, these results establish a model of fear memory formation in which intrinsic excitability determines neuronal selection, whereas learning-related encoding is governed by synaptic plasticity.


Subject(s)
Basolateral Nuclear Complex/metabolism , Conditioning, Classical/physiology , Cytoskeletal Proteins/metabolism , Fear/physiology , Memory/physiology , Nerve Tissue Proteins/metabolism , Acoustic Stimulation/adverse effects , Action Potentials/drug effects , Action Potentials/genetics , Animals , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Basolateral Nuclear Complex/cytology , Central Nervous System Stimulants/pharmacology , Choline O-Acetyltransferase/metabolism , Cytoskeletal Proteins/genetics , Glutamate Decarboxylase/metabolism , In Vitro Techniques , Luminescent Proteins/genetics , Luminescent Proteins/metabolism , Mice , Mice, Inbred C57BL , Nerve Tissue Proteins/genetics , Neurons/physiology , Patch-Clamp Techniques , Phosphopyruvate Hydratase/metabolism , Picrotoxin/pharmacology , Proto-Oncogene Proteins c-fos/metabolism
4.
Neuroscience ; 116(3): 685-94, 2003.
Article in English | MEDLINE | ID: mdl-12573711

ABSTRACT

Amyotrophic lateral sclerosis is a lethal neurodegenerative disorder involving motoneuron loss in the cortex, brainstem and spinal cord, resulting in progressive paralysis. Aberrant neurotrophin signalling via the low affinity neurotrophin receptor p75 has been suggested to be involved in the motoneuron death by the activation of apoptotic pathways. In order to investigate the involvement of neurotrophin receptor p75 in the amyotrophic lateral sclerosis related motoneuron degeneration process, we have studied the expression of this receptor in the spinal cord of transgenic mice carrying a mutated human Cu, Zn superoxide dismutase gene. Mutations in the superoxide dismutase gene are one of the genetic causes for familiar amyotrophic lateras sclerosis and human superoxide dismutase-1 transgenic mice develop symptoms and pathology similar to those in human amyotrophic lateras sclerosis. Our study shows that in these mice, spinal motoneurons, which normally do not contain the neurotrophin receptor p75 receptor, express this receptor during the progress of the disease. Expression of the neurotrophin receptor p75 receptor coincides with the expression of activating transcription factor 3, a member of the activating transcription factor/cyclic AMP family of stress transcription factors. Only a minority of these spinal motoneurons actually showed co-expression of neurotrophin receptor p75 with caspase-3 activity, suggesting that expression of the neurotrophin receptor p75 receptor is not directly related to the execution phase of the apoptosis process.


Subject(s)
Amyotrophic Lateral Sclerosis/metabolism , Disease Models, Animal , Motor Neurons/metabolism , Receptors, Nerve Growth Factor/biosynthesis , Amyotrophic Lateral Sclerosis/genetics , Animals , Female , Gene Expression Regulation/physiology , Male , Mice , Mice, Transgenic , Receptor, Nerve Growth Factor , Receptors, Nerve Growth Factor/deficiency , Receptors, Nerve Growth Factor/genetics , Superoxide Dismutase/deficiency , Superoxide Dismutase/genetics , Superoxide Dismutase-1
5.
Acta Neuropathol ; 102(4): 293-305, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11603803

ABSTRACT

Cytosolic Cu/Zn superoxide dismutase (SOD1) is a ubiquitous small cytosolic metalloenzyme, which catalyses the conversion of superoxide anion to hydrogen peroxide. Mutations in the SOD1 gene cause a familial form of amyotrophic lateral sclerosis (fALS). The mechanism by which mutant SOD1s cause the degeneration of motor neurons is not understood. Transgenic mice expressing multiple copies of fALS-mutant SOD1s develop an ALS-like motor neuron disease. Vacuolar degeneration of mitochondria has been identified as the main pathological feature associated with motor neuron death and paralysis in several lines of fALS-SOD1 mice. Using confocal and electron microscopy we show that mutant SOD1 is present at a high concentration in vacuolated mitochondria, where it colocalises with cytochrome c. Mutant SOD1 is also present in mildly swollen mitochondria prior to the appearance of vacuoles, suggesting that the leakage or translocation of mutant human SOD1 in mitochondria may be the primary event triggering their further degeneration. Vacuolated mitochondria containing SOD1 also occur in transgenic mice expressing a high concentration of wildtype human SOD1. In sum, our data suggest that both fALS-mutant and wild-type SOD1 may cross the mitochondrial outer membrane, and by doing so induce the degeneration of these mitochondria.


Subject(s)
Amyotrophic Lateral Sclerosis/metabolism , Amyotrophic Lateral Sclerosis/pathology , Mitochondria/enzymology , Superoxide Dismutase/genetics , Superoxide Dismutase/metabolism , Animals , Cytochrome c Group/analysis , Cytochrome c Group/metabolism , Female , Male , Mice , Mice, Inbred Strains , Mice, Transgenic , Microscopy, Confocal , Microscopy, Immunoelectron , Mitochondria/pathology , Mitochondria/ultrastructure , Mitochondrial Swelling , Motor Neurons/metabolism , Motor Neurons/pathology , Oxidative Stress/physiology , Superoxide Dismutase/analysis , Vacuoles/metabolism
6.
Neurobiol Dis ; 7(6 Pt B): 623-43, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11114261

ABSTRACT

Cytosolic Cu/Zn superoxide dismutase (SOD1) is a ubiquitous small cytosolic metalloenzyme that catalyzes the conversion of superoxide anion to hydrogen peroxide (H(2)O(2)). Mutations in the SOD1 gene cause a familial form of amyotrophic lateral sclerosis (fALS). The mechanism by which mutant SOD1s causes ALS is not understood. Transgenic mice expressing multiple copies of fALS-mutant SOD1s develop an ALS-like motoneuron disease resembling ALS. Here we report that transgenic mice expressing a high concentration of wild-type human SOD1 (hSOD1(WT)) develop an array of neurodegenerative changes consisting of (1) swelling and vacuolization of mitochondria, predominantly in axons in the spinal cord, brain stem, and subiculum; (2) axonal degeneration in a number of long fiber tracts, predominantly the spinocerebellar tracts; and (3) at 2 years of age, a moderate loss of spinal motoneurons. Parallel to the development of neurodegenerative changes, hSOD1(WT) mice also develop mild motor abnormalities. Interestingly, mitochondrial vacuolization was associated with accumulation of hSOD1 immunoreactivity, suggesting that the development of mitochondrial pathology is associated with disturbed SOD1 turnover. In this study we also crossed hSOD1(WT) mice with a line of fALS-mutant SOD1 mice (hSOD1(G93A)) to generate "double" transgenic mice that express high levels of both wild-type and G93A mutant hSOD1. The "double" transgenic mice show accelerated motoneuron death, earlier onset of paresis, and earlier death as compared with hSOD1(G93A) littermates. Thus in vivo expression of high levels of wild-type hSOD1 is not only harmful to neurons in itself, but also increases or facilitates the deleterious action of a fALS-mutant SOD1. Our data indicate that it is important for motoneurons to control the SOD1 concentration throughout their processes, and that events that lead to improper synthesis, transport, or breakdown of SOD1 causing its accumulation are potentially dangerous.


Subject(s)
Amyotrophic Lateral Sclerosis/pathology , Axons/pathology , Mitochondria/pathology , Motor Neurons/pathology , Superoxide Dismutase/biosynthesis , Aging/metabolism , Amyotrophic Lateral Sclerosis/enzymology , Amyotrophic Lateral Sclerosis/genetics , Animals , Axons/ultrastructure , Brain Stem/enzymology , Brain Stem/pathology , Cell Death , Crosses, Genetic , Disease Models, Animal , Disease Progression , Electron Transport Complex IV/metabolism , Gene Dosage , Hippocampus/enzymology , Hippocampus/pathology , Humans , JNK Mitogen-Activated Protein Kinases , Mice , Mice, Inbred Strains , Mice, Transgenic , Mitochondria/enzymology , Mitochondria/ultrastructure , Mitogen-Activated Protein Kinases/metabolism , Motor Neurons/enzymology , Motor Neurons/ultrastructure , Neurofilament Proteins/metabolism , Spinal Cord/enzymology , Spinal Cord/pathology , Spinocerebellar Tracts/enzymology , Spinocerebellar Tracts/pathology , Superoxide Dismutase/genetics , Superoxide Dismutase-1
7.
Brain Res ; 868(2): 165-75, 2000 Jun 23.
Article in English | MEDLINE | ID: mdl-10854569

ABSTRACT

Apolipoprotein E (apoE) is a major apolipoprotein in the central nervous system (CNS) that may play a role in various CNS disorders. ApoE is primarily localised in astrocytes, but neuronal apoE mRNA expression has been demonstrated in normal and diseased human brain, as well as in ischaemic rat brain. To obtain further insight into the role of apoE in neuronal degeneration in the CNS and conditions of neuronal apoE localisation, we have investigated in mice the distribution of apoE following neuronal injury induced by kainic acid (n=35, 25 or 35 mg kainic acid/kg BW). Consecutive series of brain sections were immunostained for apoE and markers for astroglia (GFAP) and microglia/macrophage cells (CR3). Degenerating neurones were identified with a silver-degeneration staining technique. The intensity and cellular distribution of apoE-immunoreactivity (apoE-ir) was dependent on the severity of neuronal injury. Mice that developed mild neuronal degeneration, restricted to a subset of neurones in the hippocampus, showed increased apoE-ir in astrocytes concomitant with increased GFAP-ir and mild microgliosis. In these mice, no neuronal apoE-ir was detected. In contrast, mice developing severe neuronal injury in the hippocampus - frequently also showing degeneration in other brain regions including cortex, thalamus, striatum and amygdala - showed intense apoE-ir in degenerating neurones. Surrounding the lesion, apoE-ir was increased in neuropil recurrently whereas GFAP-ir astrocytes disappeared. Thus, in mice apoE accumulates in degenerating neurones in conditions of severe neuronal injury putatively in association with disruption of the glial network.


Subject(s)
Apolipoproteins E/metabolism , Nerve Degeneration/physiopathology , Neurons/metabolism , Animals , Behavior, Animal/drug effects , Behavior, Animal/physiology , Brain/metabolism , Brain/pathology , Brain/physiopathology , Kainic Acid/pharmacology , Male , Mice , Mice, Inbred Strains , Nerve Degeneration/chemically induced , Nerve Degeneration/pathology , Neurodegenerative Diseases/etiology , Neurodegenerative Diseases/pathology , Neurodegenerative Diseases/physiopathology , Neuroglia/metabolism , Neuroglia/pathology , Neurons/pathology , Neurotoxins/pharmacology , Seizures/chemically induced , Seizures/pathology , Seizures/physiopathology
10.
J Neurocytol ; 27(5): 303-27, 1998 Jun.
Article in English | MEDLINE | ID: mdl-9923978

ABSTRACT

Unipolar brush cells (UBCs) are a class of small neurons that are densely concentrated in the granular layers of the vestibulocerebellar cortex and dorsal cochlear nucleus. The UBCs form giant synapses with individual mossy fibre rosettes on the dendrioles which make up their brush formations and are provided with numerous, unusual non-synaptic appendages. In accord with the glutamatergic nature of mossy fibres, our previous post-embedding immunocytochemical studies indicated that various ionotropic glutamate receptor subunits are localized at the post-synaptic densities of the giant synapses, whereas the non-synaptic appendages are immunonegative. On the contrary, the metabotropic glutamate receptors mGluR1alpha and mGluR2/3 are situated at the non-synaptic appendages and are lacking at the post-synaptic densities. Other authors, however, have shown that antibodies to these metabotropic receptors stain both appendages and post-synaptic densities. In the present study, we have re-evaluated the distribution of metabotropic glutamate receptors in the UBCs of the cerebellum and the cochlear nuclear complex by light and electron microscopic pre-embedding immunocytochemistry with subtype-specific antibodies. We confirm that UBCs dendritic brushes are densely immunostained by antibody to mGluR1alpha particularly in the cerebellum and that antibody to mGluR2/3 labels at least a percentage of the UBC brushes in both the cerebellum and cochlear nuclei. At the ultrastructural level, it appears that mGluR1alpha and mGluR2/3 immunoreactivities are not associated with the post-synaptic densities of the giant mossy fibre-UBC synapses, but instead are concentrated on the non-synaptic appendages of the cerebellar UBCs. The non-synaptic appendages, therefore, may be an important avenue for regulating the excitability of UBCs and mediating glutamate effects on their still unknown intracellular signal transduction cascades. We also show that the pre-synaptic densities of UBC dendrodendritic junctions are mGluR2/3 positive. As previously demonstrated, antibodies to mGluR1alpha and mGluR2/3 label subsets of Golgi cells. Antibody to mGluR5 does not stain UBCs in the cerebellum and cochlear nucleus and reveals the somatodendritic compartment of Golgi cells situated in the core of the cerebellar granular layer, whilst cochlear nucleus Golgi cells are mGluR5 negative.


Subject(s)
Cerebellar Cortex/cytology , Cochlear Nucleus/cytology , Neurons/chemistry , Receptors, Metabotropic Glutamate/analysis , Animals , Axons/chemistry , Cell Membrane/chemistry , Dendrites/chemistry , Female , Immunohistochemistry , Male , Microscopy, Immunoelectron , Neurons/ultrastructure , Purkinje Cells/chemistry , Purkinje Cells/ultrastructure , Rats , Rats, Sprague-Dawley , Synapses/chemistry
11.
Prog Brain Res ; 114: 67-96, 1997.
Article in English | MEDLINE | ID: mdl-9193139

ABSTRACT

We have studied the source and ultrastructural characteristics of ChAT-immunoreactive fibers in the cerebellum of the rat, and the distribution of muscarinic and nicotinic receptors in the cerebellum of the rat, rabbit, cat and monkey, in order to define which of the cerebellar afferents may use ACh as a neurotransmitter, what target structures are they, and which cholinergic receptor mediate the actions of these pathways. Our data confirm and extend previous observations that cholinergic markers occur at relatively low density in the cerebellum and show not only interspecies variability, but also heterogeneity between cerebellar lobules in the same species. As previously demonstrated by Barmack et al. (1992a,b), the predominant fiber system in the cerebellum that might use ACh as a transmitter or a co-transmitter is formed by mossy fibers originating in the vestibular nuclei and innervating the nodulus and ventral uvula. Our results show that these fibers innervate both granule cells and unipolar brush cells, and that the presumed cholinergic action of these fibers most likely is mediated by nicotinic receptors. In addition to cholinergic mossy fibers, the rat cerebellum is innervated by beaded ChAT-immunoreactive fibers. We have demonstrated that these fibers originate in the pedunculopontine tegmental nucleus (PPTg), the lateral paragigantocellular nucleus (LPGi), and to a lesser extent in various raphe nuclei. In both the cerebellar cortex and the cerebellar nuclei these fibers make asymmetric synaptic junctions with small and medium-sized dendritic profiles. Both muscarinic and nicotinic receptor could mediate the action of these diffuse beaded fibers. In the cerebellar nuclei the beaded cholinergic fibers form a moderately dense network, and could in principle have a significant effect on neuronal activity. For instance, the cholinergic fibers arising in the PPTg may modulate the excitability of the cerebellonuclear neurons in relation to sleep and arousal (e.g. McCormick, 1989). Studies on the distribution of cholinergic markers in the cerebellum have proven valuable besides the issue whether cholinergic mechanism play a role in the cerebellar circuitry, because they illustrate a complexity of the cerebellar anatomy that extends beyond its regular trilaminar and foliar arrangement. For instance, AChE histochemistry has been shown to preferentially stain the borders of white matter compartments (the 'raphes', Voogd, 1967), and therefore is useful in topographical analysis of the cortico-nuclear and olivocerebellar projections (Hess and Voogd, 1986; Tan et al., 1995; Voogd et al., 1996; see Voogd and Ruigrok, 1997, this Volume). ChAT-immunoreactivity, at least in rat, appears to be a good marker to outline the morphological heterogeneity of mossy fibers, and m2-immunocytochemistry could be used to label (subpopulations of) Golgi cells, subsets of mossy fibers and, in the rabbit, a specific subset of Purkinje cells (Jaarsma et al., 1995).


Subject(s)
Acetylcholine/physiology , Afferent Pathways/physiology , Cerebellum/physiology , Choline O-Acetyltransferase/analysis , Nerve Fibers/ultrastructure , Neurons/physiology , Receptors, Muscarinic/analysis , Receptors, Nicotinic/analysis , Afferent Pathways/cytology , Animals , Cats , Cerebellum/cytology , Haplorhini , Nerve Fibers/physiology , Neurotransmitter Agents/physiology , Rabbits , Rats , Species Specificity
12.
Prog Brain Res ; 114: 131-50, 1997.
Article in English | MEDLINE | ID: mdl-9193142

ABSTRACT

The unipolar brush cell (UBC) is a novel type of small neuron that is characterized by sets of morphological and chemical phenotypes. UBCs occur in the granular layer of the mammalian cerebellar cortex, particularly in folia of the vestibulocerebellum, and in the granule cell domains of the dorsal cochlear nucleus. The UBC is characterized by a single dendrite that terminates with a brush-like tip of dendrioles. The soma, the dendritic stem, and especially the dendrioles emit short, non-synaptic appendages. The dendrioles represent the main synaptic apparatus of the UBC and articulate tightly with a single mossy fiber rosette forming a glomerular array characterized by an extraordinarily extensive synaptic contact. Electron microscopic and electrophysiological observations indicate that the unusual synaptic ultrastructure may produce entrapment of neurotransmitter in the synaptic cleft. While ionotropic glutamate receptors are enriched in correspondence of the postsynaptic density, metabotropic glutamate receptors are situated extrasynaptically and are particularly enriched at the appendages, which usually do not bear synaptic junctions. Some of the UBCs receive their input from choline acetyltransferase-positive mossy rosettes originating from the vestibular nuclei, suggesting that ACh and glutamate are co-released at these synapses. The UBC brush occupies a glomerulus where granule cell dendrites are intermixed with the UBC dendrioles, both of which receive synapses from the same mossy fiber rosette and portions of the Golgi axonal plexus. In addition, the dendrioles are presynaptic to granule cell dendrites, forming dendrodendritic contacts that display features of excitatory synapses. Branches of the UBC axon in the granular layer bear large endings resembling mossy fibers. The UBCs may represent an extraordinary device for feedforward, excitatory links along the mossy fiber pathways of cerebellum and dorsal cochlear nucleus.


Subject(s)
Cerebellum/cytology , Cerebellum/physiology , Cochlear Nucleus/cytology , Cochlear Nucleus/physiology , Neurons/cytology , Neurons/physiology , Animals , Humans , Mammals , Models, Neurological , Nerve Fibers/physiology , Nerve Fibers/ultrastructure , Neurotransmitter Agents/analysis , Neurotransmitter Agents/physiology , Synapses/physiology , Synapses/ultrastructure
13.
J Neurocytol ; 25(12): 829-42, 1996 Dec.
Article in English | MEDLINE | ID: mdl-9023728

ABSTRACT

A subset of cerebellar mossy fibres is rich in choline acetyltransferase, the rate-limiting enzyme for the synthesis of acetylcholine. These choline acetyltransferase-positive mossy fibres are concentrated in the vestibulocerebellum and originate predominantly from the medial vestibular nucleus. The granular layer of the vestibulocerebellum is also enriched in unipolar brush cells, an unusual type of small neuron that form giant synapses with mossy fibres. In this immunocytochemical light and electron microscopic study, we explored whether choline acetyltransferase-positive mossy fibres innervate unipolar brush cells of the rat cerebellum. We utilized monoclonal antibodies to rat choline acetyltransferase of proven specificity, and immunoperoxidase procedures with 3,3'-diaminobenzidine tetrahydrochloride as the chromogen. A high density of choline acetyltransferase-positive fibres occurred in the nodulus and ventral uvula, where they showed an uneven, zonal distribution. Immunostained mossy fibre rosettes contained high densities of round synaptic vesicles and mitochondria. They formed asymmetric synaptic junctions with dendritic profiles of both granule cells and unipolar brush cells. The synaptic contacts between choline acetyltransferase-immunoreactive mossy fibres and unipolar brush cells were very extensive, and did not differ from synapses of choline acetyltransferase-negative mossy fibres with unipolar brush cells. Analysis of a total area of 1.25 mm2 of the nodulus from three rats revealed that 14.2% of choline acetyltransferase-immunoreactive mossy fibre rosettes formed synapses with unipolar brush cells profiles. Choline acetyltransferase-positive rosettes accounted for 21.7% of the rosettes forming synapses with unipolar brush cells. Thus, the present data demonstrate that unipolar brush cells are innervated by a heterogeneous population of mossy fibres, and that some unipolar brush cells receive cholinergic synaptic input from the medial vestibular nucleus. The ultrastructure of these synapses is compatible with the possibility that choline acetyltransferase-positive mossy fibres co-release acetylcholine and glutamate. As the granular layer of the vestibulocerebellum contains nicotinic binding sites, the choline acetyltransferase-positive mossy fibres may be a model for studying nicotinic neurotransmission in the CNS.


Subject(s)
Cerebellum/cytology , Cerebellum/enzymology , Choline O-Acetyltransferase/analysis , Nerve Fibers/ultrastructure , Neurons/cytology , Receptors, Nicotinic/physiology , Synaptic Transmission , Animals , Antibodies, Monoclonal , Cerebellum/ultrastructure , Cytoplasmic Granules/enzymology , Cytoplasmic Granules/ultrastructure , Female , Immunoenzyme Techniques , Immunohistochemistry , Male , Microscopy, Immunoelectron , Nerve Fibers/enzymology , Neurons/enzymology , Neurons/ultrastructure , Rats , Rats, Sprague-Dawley , Rats, Wistar , Receptors, Nicotinic/analysis , Synapses/physiology , Synapses/ultrastructure
14.
Neurosci Lett ; 219(3): 179-82, 1996 Nov 29.
Article in English | MEDLINE | ID: mdl-8971809

ABSTRACT

Transgenic mice carrying amyotrophic lateral sclerosis (ALS)-linked superoxide dismutase 1 (SOD1) mutations develop a motoneuron disease resembling human ALS. c-Jun is a transcription factor frequently induced in injured neurons. In this study we have examined the distribution of c-Jun-immunoreactivity in the brainstem and spinal cord of transgenic SOD1 mice with a glycine 93 alanine (G93A) mutation. In non-transgenic littermates c-Jun immunostaining was predominantly situated in motoneurons. The number of c-Jun immunoreactive motoneuron was reduced in SOD1(G93A) mice due to pronounced loss of motoneurons. In SOD1(G93A) mice, however, c-Jun-immunoreactivity was strongly induced in neurons in the intermediate zone (Rexed's laminae V-VIII and X) of the spinal cord and throughout the brainstem reticular formation. These findings are of interest since increased levels of c-jun also have been found in the intermediate zone of the spinal cord of ALS patients. This c-Jun may be involved in the neurodegenerative processes both in ALS and in motoneuron disease in SOD1(G93A) mice.


Subject(s)
Amyotrophic Lateral Sclerosis/metabolism , Brain Stem/metabolism , Neurons/metabolism , Proto-Oncogene Proteins c-jun/metabolism , Spinal Cord/metabolism , Amyotrophic Lateral Sclerosis/genetics , Animals , Brain Stem/cytology , Immunologic Techniques , Isoenzymes/genetics , Mice , Mice, Transgenic , Motor Neurons/metabolism , Mutation , Spinal Cord/cytology , Superoxide Dismutase/genetics , Tissue Distribution
15.
J Chem Neuroanat ; 9(4): 241-59, 1995 Dec.
Article in English | MEDLINE | ID: mdl-8719274

ABSTRACT

Recent studies on the effects of intrafloccular injections of muscarinic agonists and antagonists on compensatory eye movements in rabbit, indicate that muscarinic receptors may play a modulatory role in the rabbit cerebellar circuitry. It was previously demonstrated by Neustadt et al. (1988), that muscarinic receptors in rabbit cerebellar cortex are distributed into alternating longitudinal zones of very high and very low receptor density. In the present study, the zonal and cellular distribution of muscarinic receptors in the rabbit cerebellar cortex is investigated in detail using in vitro ligand autoradiography with the non-selective high-affinity antagonist [3H]quinuclidinyl benzilate (QNB), and the M2-specific antagonist [3H]AF-DX384, and immunocytochemistry with a monoclonal antibody specific for the cloned m2 muscarinic receptor protein. [3H]QNB and [3H]AF-DX384 binding sites and m2-immunoreactivity had similar overall distributions: dense labeling occurred in the dendritic arbors of a subset of Purkinje cells that are organized into parasagittal bands. A high level of muscarinic receptor labeling was also observed in a thin substratum of the molecular layer immediately above the Purkinje cell layer of the vestibulo-cerebellar lobules, i.e. the nodulus, the ventral uvula and the flocculus. Labeling in this stratum was associated with densely packed fibres, which were putatively identified as parallel fibres. Also Golgi cells, which were localized in part in the molecular layer, and a subset of mossy fibre rosettes, primarily concentrated in lobule VI, were immunoreactive for the m2 receptor. The parasagittal band of labeled Purkinje cell dendrites were most prominent in the anterior lobe (lobules I-V), in crus 1 and 2, in the flocculus, the ventral paraflocculus and the rostral folium of the nodulus. In other lobules, only infrequent Purkinje cells contained muscarinic receptors. The parasagittal organisation of muscarinic receptors differed from that of zebrin I, a Purkinje cell-specific protein which is often used as a marker of parasagittal parcelation of the cerebellar cortex. In the anterior lobe, however, there was a partial correspondence between muscarinic receptor and zebrin I bands. In the flocculus the distribution of muscarinic-receptor-positive Purkinje cells was related to the distinct white matter compartments as revealed with acetylcholinesterase (AChE) histochemistry. Muscarinic receptor-containing Purkinje cells were located primarily in the floccular zone 1, which is implicated in the control of eye movements about a horizontal axis. In order to relate the distribution of muscarinic receptor labeling to that of cholinergic nerve terminals, [3H]QNB binding sites and sodium-dependent [3H]hemicholinium-3 binding were compared. Sodium-dependent [3H]hemicholinium-3 binding sites mainly occurred in the granule cell layer of the vestibulo-cerebellum, which corresponds well with the distribution of the acetylcholine synthesizing enzyme, choline acetyltransferase (ChAT). However, sodium-dependent [3H]hemicholinium binding complemented, rather than co-localized with, muscarinic receptors which were primarily distributed in the molecular layer of the lobules of the vestibulo-cerebellar lobules. Their functional significance is puzzling, since their distribution does not correspond to that of markers of cholinergic innervation.


Subject(s)
Cerebellar Cortex/metabolism , Receptors, Muscarinic/metabolism , Acetylcholinesterase/metabolism , Animals , Autoradiography , Cerebellar Cortex/anatomy & histology , Female , Hemicholinium 3 , Immunohistochemistry , Male , Muscarinic Antagonists , Nerve Tissue Proteins/metabolism , Parasympatholytics , Pirenzepine/analogs & derivatives , Purkinje Cells/metabolism , Quinuclidinyl Benzilate , Rabbits , Sodium/physiology
16.
J Comp Neurol ; 357(1): 145-60, 1995 Jun 19.
Article in English | MEDLINE | ID: mdl-7673463

ABSTRACT

The present study provides a survey of the immunolocalization of ionotropic glutamate receptor subunits throughout the rat and cat cerebellar cortex, with emphasis on the unipolar brush cell (UBC), a hitherto neglected cerebellar cell that is densely concentrated in the granular layer of the vestibulocerebellum and that forms giant synapses with mossy fibers. An array of nine previously characterized antibodies has been used, each of which stained a characteristic profile of cerebellar cells. The UBCs of both rat and cat were strongly immunostained by an antibody against the alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionate (AMPA) receptor subunits, GluR2 and GluR3; were moderately immunostained by a monoclonal antibody to kainate receptor subunits, GluR5/6/7; were weakly immunostained by antibodies to NR1 subunits; and were not stained by antibodies to GluR1, GluR4, GluR6/7, KA-2, and NR2A/B. Postsynaptic densities of the giant mossy fiber-UBC synapses were GluR2/3, GluR5/6/7, and NR1 immunoreactive. The other cerebellar neurons were all immunolabeled to some extent with the GluR2/3 and NR1 antibodies. In addition, Purkinje cells were immunopositive for GluR1 and GluR5/6/7; granule cells were immunopositive for GluR5/6/7, GluR6/7, KA-2, and NR2A/B. The Golgi-Bergmann glia was densely stained by GluR1 and GluR4 antibodies, whereas astrocytes of the granular layer were stained by the GluR4 antiserum. Data provided herein may guide further electrophysiological and pharmacological studies of cerebellar cells in general and the UBCs in particular.


Subject(s)
Cats/metabolism , Cerebellar Cortex/chemistry , Nerve Fibers/chemistry , Rats, Sprague-Dawley/metabolism , Receptors, Glutamate/analysis , Synapses/chemistry , Afferent Pathways , Animals , Cerebellar Cortex/cytology , Female , Immunohistochemistry , Male , Microscopy , Microscopy, Electron , Rats , Receptors, AMPA/analysis , Receptors, Kainic Acid/analysis , Receptors, N-Methyl-D-Aspartate/analysis
17.
J Neurol Sci ; 127(2): 230-3, 1994 Dec 20.
Article in English | MEDLINE | ID: mdl-7707082

ABSTRACT

The tissue concentrations of two related amino acid derivatives, N-acetylaspartate (NAA) and N-acetylaspartylglutamate (NAAG) were determined in autopsy hippocampus, amygdala, cerebellar cortex and olfactory bulb of Alzheimer's disease patients and age-matched non-demented controls, using reverse-phase HPLC and fluorescence detection after precolumn derivatisation with the fluorophore 2-aminoanthracene. In Alzheimer's disease, NAA and NAAG concentrations were significantly reduced in the hippocampus (by 38 and 24%) and the amygdala (by 28 and 22%), but not in the olfactory bulb and the cerebellar cortex. These results indicate that the concentrations of NAA and NAAG are selectively decreased in brain areas affected by pathology in Alzheimer's disease.


Subject(s)
Alzheimer Disease/metabolism , Aspartic Acid/analogs & derivatives , Brain Chemistry/physiology , Dipeptides/metabolism , Histamine H1 Antagonists/metabolism , Aged , Aged, 80 and over , Amygdala/metabolism , Aspartic Acid/metabolism , Cerebellar Cortex/metabolism , Chromatography, High Pressure Liquid , Female , Hippocampus/metabolism , Humans , Male , Middle Aged , Olfactory Bulb/metabolism , Spectrometry, Fluorescence
18.
J Neurosci Methods ; 46(2): 133-8, 1993 Feb.
Article in English | MEDLINE | ID: mdl-7682638

ABSTRACT

The novel high-affinity competitive NMDA receptor antagonist, CGP39653, is employed as radioligand to autoradiographically label the NMDA receptor in rat and human brain. Glutamate dehydrogenase (GlDH; E.C.1.4.1.3) was added to the incubation buffer to degrade residual endogenous L-glutamate, which was not entirely removed from the section after the prewashing step and interfered with [3H]CGP39653 binding. At 20 nM [3H]CGP39653, GLDH increased specific binding as much as 5 times, depending on the dose of GlDH and the presence of NAD+, and hydrazine. Scatchard plots of binding data revealed that this increase was due to a decrease of the KD from 148 nM to 33 nM in the absence and the presence of GlDH, respectively. Addition of GlDH in the NMDA receptor autoradiographic assay may be of importance in quantitative studies with human brain tissue which may contain variable levels of endogenous L-glutamate.


Subject(s)
2-Amino-5-phosphonovalerate/analogs & derivatives , Glutamate Dehydrogenase/pharmacology , Receptors, N-Methyl-D-Aspartate/metabolism , 2-Amino-5-phosphonovalerate/pharmacokinetics , 6-Cyano-7-nitroquinoxaline-2,3-dione , Animals , Aspartic Acid/pharmacokinetics , Autoradiography , Brain/anatomy & histology , Brain Chemistry/drug effects , Glutamates/pharmacokinetics , Glutamic Acid , Ibotenic Acid/analogs & derivatives , Ibotenic Acid/metabolism , In Vitro Techniques , Male , Quinoxalines/pharmacology , Rats , Rats, Wistar , Receptors, AMPA , Receptors, Glutamate/drug effects , Receptors, N-Methyl-D-Aspartate/antagonists & inhibitors , Receptors, N-Methyl-D-Aspartate/drug effects , alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid
19.
J Auton Nerv Syst ; 41(1-2): 157-76, 1992 Nov.
Article in English | MEDLINE | ID: mdl-1491112

ABSTRACT

The aim of the present study was to explore the neuroanatomic network that underlies the cardiovascular responses of reticular formation origin in the region of the cuneiform nucleus (CNF). The study was performed in urethane anesthetized male Wistar rats. The left iliac artery was supplied with a catheter for the measurement of systemic blood pressure. Low intensity electrical stimulation of the mesencephalic reticular formation (MRF) in the vicinity of the CNF always resulted in pressor and bradycardiac responses, whereas stimulation in the parabrachial nucleus (PB) and Kölliker-Fuse nucleus (KF) led to a pressor response and a small tachycardiac response. The cuneiform area may be placed in the center of a circuit that serves a specific autonomic response pattern to stress: parallel activation of the sympathetic (pressor response) and parasympathetic limb (bradycardia). The efferent connections of the effective stimulation sites in the MRF and the CNF area, were investigated by anterograde tracing with the lectin Phaseolus vulgaris leucoagglutine (PHA-L). The CNF sends descending fibers to the gigantocellular reticular nuclei (GI), the motor nucleus of the vagus (DMNV) and nucleus tractus solitarius (NTS). These projections are probably involved in the bradycardiac response to stimulation. The descending pathway to the NTS/DMNV and GI may therefore be the parasympathetic limb of the circuit. Furthermore, the CNF sends ascending fibers to limbic forebrain areas and descending fibers to the PB-KF complex. The KF in its turn projects to the rostroventrolateral medullary nucleus (RVLM) and the intermediolateral cell column (IML). These latter projections are partly involved in producing the pressor response and thereby represent the sympathetic limb of the circuit. Accordingly, the transection of the descending fibers from the CNF to the PB-KF complex resulted in a decreased pressor and an increased bradycardiac response. This suggests that a baroreceptor reflex-induced bradycardia which results from blood pressure increase can be excluded as the origin of the stimulation-induced bradycardia, and that the pressor and bradycardiac responses are two independent moieties. It cannot be excluded that ascending fibers from the CNF are also involved in producing the pressor response. On the basis of the present physiological and neuroanatomical study, a brain circuit has been proposed in which the cuneiform nucleus has a central position. The described brain circuit may serve a passive coping strategy to novel, painful or threatening stimuli during which the animals show orientation/attention or freezing behavior accompanied by a bradycardiac and pressor response.


Subject(s)
Hemodynamics/physiology , Mesencephalon/physiopathology , Stress, Psychological/physiopathology , Adaptation, Psychological/physiology , Afferent Pathways/physiology , Animals , Axons/physiology , Blood Pressure/physiology , Efferent Pathways/physiology , Electric Stimulation , Electrodes , Heart Rate/physiology , Immunohistochemistry , Male , Mesencephalon/pathology , Phytohemagglutinins , Rats , Rats, Wistar , Reticular Formation/anatomy & histology , Reticular Formation/physiology
20.
Hippocampus ; 2(2): 143-50, 1992 Apr.
Article in English | MEDLINE | ID: mdl-1308179

ABSTRACT

Recently, Sloviter et al. reported that adrenalectomy (ADX) of young adult rats after 3 months led to a selective loss of granule neurons in the dentate gyrus (DG) and that this loss could be prevented by low doses of corticosterone. In the present study, the ADX-induced neuronal degeneration was investigated in Wistar rats, using a silver impregnation method for degenerating neurons. To examine the time course and distribution of the ADX-induced degeneration, young adult male rats were allowed to survive 2, 3, and 5 days and 1, 2, and 3 weeks after ADX. Argyrophilic neurons were present in the dentate granule cell layer on the second day following ADX. Three days after ADX, the number of argyrophilic granule neurons was much more abundant, and it increased gradually with longer post-ADX survival times. Argyrophilia was specifically confined to dentate granule cells and was accompanied by the occurrence of pyknotic nuclei as observed in adjacent cresyl violet-stained sections. There were significant differences between individual rats in quantity of argyrophilia. About one fifth of the ADX rats showed sporadic or no argyrophilia, in spite of plasma corticosterone levels below the detection limit (10 ng/mL). Sham-operated rats and ADX rats receiving corticosterone (10 mg/L) or dexamethasone (15 mg/L) in their drinking water did not display any argyrophilic neurons in the dentate gyrus. The distribution of the argyrophilia within the DG was highly characteristic with the highest number of degenerating cells in the hidden blade of the middle and the temporal thirds of the DG.(ABSTRACT TRUNCATED AT 250 WORDS)


Subject(s)
Adrenalectomy , Dexamethasone/pharmacology , Hippocampus/physiology , Nerve Degeneration , Neurons/physiology , Aging/physiology , Animals , Axons/drug effects , Axons/physiology , Axons/ultrastructure , Body Weight , Corticosterone/blood , Dendrites/drug effects , Dendrites/physiology , Dendrites/ultrastructure , Female , Hippocampus/drug effects , Hippocampus/ultrastructure , Male , Nerve Degeneration/drug effects , Neurons/drug effects , Neurons/ultrastructure , Rats , Rats, Wistar , Sex Factors , Silver , Species Specificity , Time Factors
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