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M Dragunow

Publications and source records attributed to M Dragunow.

At least 73 records · Page 4Linked to original sources
PubMed1995

In situ evidence for DNA fragmentation in Huntington's disease striatum and Alzheimer's disease temporal lobes.

To test the hypothesis that apoptosis is involved in human brain neurodegenerative disorders, we investigated whether DNA fragmentation occurs in Alzheimer's disease (AD). Huntington's disease (HD) and Parkinson's disease, as well as in temporal lobe epilepsy, using neurologically normal post-mortem human brain tissue as a control. Using in situ end labelling of DNA, we found evidence of DNA fragmentation in cells in temporal cortex and hippocampus from patients with AD and in striatum from those with HD. In contrast, only scattered DNA fragmentation positive cells were detected in the pial surfaces of some of the neurologically normal human brains. Thus, cells in the HD striatum and AD temporal cortex exhibited DNA fragmentation, suggesting that apoptosis may be involved in these disorders.

Adult↗
PubMed1995

trkC may be an inducible transcription factor target gene.

Mechanical injury to hippocampus by needle insertion induces N-methyl-D-aspartate (NMDA) receptor-dependent expression of neurotrophin receptor mRNAs (trkB, trkC) and inducible transcription factors (ITFs, i.e. Fos, c-Jun, Krox-24) in dentate gyrus neurones. To examine the possibility that de novo synthesis of ITFs was necessary for increased expression of trk mRNA we prevented protein synthesis for 5 h using the protein synthesis inhibitor, cycloheximide (4 x 10 mg kg-1, i.p., 1 h before and 1, 2 and 3 h after injury). Cycloheximide abolished expression of ITFs 4 h after injury. Cycloheximide had no effect on trkB mRNA expression but attenuated the injury-induced increase in trkC mRNA expression in dentate gyrus at 4 h by 75%. These results suggest that trkC may be an ITF target gene in vivo.

Animals↗
PubMed1995

Induction of the Krox 24 transcription factor in striosomes by a cannabinoid agonist.

The striatum is composed of two compartments arranged as a mosaic, the striosomes (patches) and the matrix, which differ in their neurochemical and neuroanatomical properties. Along with a large number of neurotransmitter-related differences, these regions have recently been shown to differ in their immediate early gene (IEG) response to indirect dopamine agonists. Both Fos and Krox 24 can be preferentially induced in the striosomal compartment of the striatum by amphetamine. This compartmentalization of response suggests that there are functionally distinct molecular signalling pathways in striosomes and matrix. This paper examines the response of Krox 24 to i.p. administration of the cannabinoid agonist CP55,940 and demonstrates a selective induction of this protein within the striosomes 2 h after drug administration. This result suggests that cannabinoid receptors may regulate striatal gene expression.

Animals↗
PubMed1995

Neuronal injury following electrically induced status epilepticus with and without adenosine receptor antagonism.

Adenosine is thought to act as an endogenous anticonvulsant and neuroprotective substance in the brain. In the present study we compared neuronal death following status epilepticus (SE) induced in the presence of 8-cyclopentyl-1,3-dimethylxanthine (8-CPT), an A1-adenosine receptor antagonist, with that following SE induced by continuous hippocampal stimulation. Hippocampal damage was characterized using selective nerve and nonnerve cell markers. Six days after SE, both models produced similar patterns of CA1 and CA3 cell loss and selective loss of parvalbumin and hilar somatostatin-immunoreactive interneurons. Calbindin D28K-immunoreactive interneuron numbers and calbindin D28K immunoreactivity in dentate granule cells remained unchanged although calbindin D28K staining was lost in damaged CA1 neurons. Neuronal injury in these areas was also accompanied by reactive gliosis and microglial proliferation, as well as the production of basic fibroblast growth factor and insulin-like growth factor-1 by astrocytes. Although hippocampal damage appeared to be more severe after SE induced in the presence of 8-CPT, this may be due to the increased severity of SE generated in this model.

Animals↗
PubMed1995

The role of inducible transcription factors in apoptotic nerve cell death.

Recent studies have shown that certain types of nerve cell death in the brain occur by an apoptotic mechanism. Researchers have demonstrated that moderate hypoxic-ischemic (HI) episodes and status epilepticus (SE) can cause DNA fragmentation as well as other morphological features of apoptosis in neurons destined to die, whereas more severe HI episodes lead to neuronal necrosis and infarction. Although somewhat controversial, some studies have demonstrated that protein synthesis inhibition prevents HI-and SE-induced nerve cell death in the brain, suggesting that apoptotic nerve cell death in the adult brain is de novo protein synthesis-dependent (i.e., programmed). The identity of the proteins involved in HI-and SE-induced apoptosis in the adult brain is unclear, although based upon studies in cell culture, a number of potential cell death and anti-apoptosis genes have been identified. In addition, a number of studies have demonstrated that inducible transcription factors (ITFs) are expressed for prolonged periods in neurons undergoing apoptotic death following HI and SE. These results suggest that prolonged expression of ITFs (in particular c-jun) may form part of the biological cascade that induces apoptosis in adult neurons. These various studies are critically discussed and in particular the role of inducible transcription factors in neuronal apoptosis is evaluated.

Adult↗
PubMed1995

Neurochemical and morphological changes associated with human epilepsy.

To date a multitude of studies into the morphology and neurochemistry of human epilepsy have been undertaken with variable, and often inconsistent, results. This review summarises these studies on a range of neurotransmitters, neuromodulators, neuropeptides and their receptors. In addition to this, novel changes in cell viability and sprouting have been identified and are discussed. Whether the alterations observed are a result of the seizures or are a contributory factor is unclear. However, it may be that following an initial insult (such as febrile convulsions, status epilepticus or head injury) secondary processes occur both of an anticonvulsant nature in an attempt to compensate for seizure activity, and in a kindling type of fashion, resulting in an increased susceptibility to seizures, leading to future seizures. Many of the alterations documented in this study probably represent one or both of these processes. Clearly no single chemical abnormality or morphological alteration is going to explain the clinically diverse disorder of epilepsy. However, by drawing together the neurochemistry and morphology of epilepsy, we may begin to understand the mechanisms involved in seizure disorders.

Brain↗
PubMed1995

Krox20 may play a key role in the stabilization of long-term potentiation.

Long-term potentiation-inducing stimulation of the perforant path was followed in dentate gyrus granule cells by a dramatic increase of mRNA and protein for Krox20, a zinc-finger-containing transcription factor. Induction of Krox20 required stimulation sufficient to induce LTP and was prevented by NMDA antagonists CPP and MK-801, which block LTP induction. Krox20 protein increased within 20 min of tetanization, was maximal between 1 and 8 h, and was still significantly elevated at 24 h after LTP induction. This prolonged appearance is in striking contrast with the more transient induction of the related molecule, Krox24. The elevation in the mRNA for Krox20 and Krox24 was of similar duration, suggesting that the Krox20 protein has a greater stability and may play a key role in the stabilization of long-term potentiation.

Animals↗
PubMed1995

Mechanisms of delayed cell death following hypoxic-ischemic injury in the immature rat: evidence for apoptosis during selective neuronal loss.

The mechanisms leading to delayed cell death following hypoxic-ischemic injury in the developing brain are unclear. We examined the possible roles of apoptosis and microglial activation in the 21-day-old rat brain following either mild (15 min) or severe (60 min) unilateral hypoxic-ischemic injury. The temporal and spatial patterns of DNA degradation were assessed using gel-electrophoresis and in-situ DNA end-labelling. Microglial activation, mitochondrial failure and cell death were examined using lectin histochemistry, 2,3,5,triphenyl-H-tetrazolium chloride (TTC) staining and acid fuchsin staining, respectively. Selective neuronal death produced by the 15 min injury was associated with the development of apoptotic morphology, DNA laddering and acidophilia from 3 days post-hypoxia. The 60 min injury accelerated this process with some cells showing signs of DNA degradation at 10 h post-hypoxia. However, in the cortex, which developed infarction after the 60 min injury, a different pattern of cell loss occurred. The DNA and mitochondria remained intact, and cells basophilic, until after 10 h post-hypoxia, then widespread necrosis developed by 24 hr. In contrast to regions of selective neuronal loss, DNA degradation was initially random (at 24 hr), with 180bp DNA ladders not detected until 3 days post-hypoxia. There was no morphological evidence of apoptosis. Microglial activation coincided with the onset of DNA degradation in regions of selective neuronal loss but not infarction, suggesting a possible role in selective neuronal death. The results suggest that cortical infarction, which was delayed for at least 10 h, was necrotic, and occurred independently of microglial activation and apoptosis. In contrast, selective neuronal death was apoptotic.

Animals↗
PubMed1995

Analysis of the decremental nature of LTP in the dentate gyrus.

The persistence of long-term potentiation (LTP) in the dentate gyrus was compared for two tetanization protocols: 50 trains on one day, or 50 trains on 5 consecutive days. LTP induction was significantly greater in the 250 train condition, but the LTP decay rate over weeks was similar between conditions. The decay of LTP could not be accounted for by deterioration of the preparation. Successive days of stimulation caused repetitive induction of immediate early genes, but did not prolong LTP, suggesting that either the effects of gene expression on LTP stabilization had saturated, or that these genes play other roles in synaptic plasticity.

Animals↗
PubMed1995

Prolonged expression of Fos-related antigens, Jun B and TrkB in dopamine-denervated striatal neurons.

Previous studies have demonstrated that striatal dopamine-denervation leads to a long-term increase in Fos-related antigen(s) (FRA's) in striatal neurons. Because Fos-family proteins bind to DNA by dimerizing to Jun-family proteins we investigated the expression of Jun B protein 2 weeks and 1 month after striatal dopamine-denervation, produced by medial forebrain bundle transection. We also investigated the effects of this lesion on TrkB-immunoreactivity in the striatum. FRA's (as previously reported) and Jun B were expressed in striatal neurons following dopamine-denervation, and in addition, there was an increase in expression of TrkB in the striatum on the dopamine-denervated side. These results show that striatal dopamine depletion leads to a long-term up-regulation of FRA's and Jun B in the striatum, and this may be related to other biochemical changes previously reported to occur in striatal neurons (e.g.: D2-dopamine receptor up-regulation) after dopamine depletion. In addition, FRA and Jun B expression may induce increased production of TrkB after dopamine-denervation.

Animals↗
PubMed1995

Clusterin accumulates in dying neurons following status epilepticus.

Clusterin is a protein that has been implicated in cell death and remodelling in a number of different tissues. To further investigate the role of clusterin in nerve cell death its expression was measured in the rat brain at various times after status epilepticus (SE) induced by 1 h of hippocampal stimulation, by using in situ hybridization, immunocytochemistry, and immunoblotting. SE lead to a dramatic time-dependent increase in clusterin mRNA in non-nerve cells resembling astrocytes in the hippocampus beginning after 24 h. There was also an earlier induction of clusterin mRNA in dentate granule cells, that survive SE. Only a low mRNA signal was observed over the CA1 pyramidal cells, which die after SE. In contrast to these mRNA results, massive clusterin-like immunoreactivity was observed in CA1 pyramidal cells and dentate hilar neurons (and both of these neuronal populations die after SE), but not in dentate granule cells. We speculate that astrocytes produce clusterin after SE and that the clusterin is then secreted and taken up by hippocampal neurons destined to die. Thus, the role of clusterin in nerve cell death/ regeneration warrants further investigation.

Animals↗
PubMed1995

GABA and GABAA receptor changes in the substantia nigra of the rat following quinolinic acid lesions in the striatum closely resemble Huntington's disease.

GABA and GABAA receptors have been studied in the substantia nigra of the rat following quinolinic acid lesions in the striatum. The regional distribution of GABA and GABAA receptors was investigated using immunohistochemical techniques with monoclonal antibodies to GABA and to the beta 2.3 subtypes of the GABAA receptor complex. The distribution, density and cellular localization of GABAA receptors were studied using quantitative receptor autoradiography and 6-hydroxydopamine-induced degeneration of dopaminergic pars compacta neurons. The subunit configuration of GABAA receptors was investigated using in situ hybridization histochemistry and subunit subtype-specific oligonucleotide probes. The results showed that in the normal substantia nigra GABA and GABAA receptors were mainly localized within the pars reticulata. GABAA receptors were mainly of the BZI variety, had a subunit subtype configuration that included alpha 1 and beta 2.3 subtypes, and showed a rostrocaudal gradient in the density of receptors; the density of receptors in the caudal third was 56% higher than that in the rostral third of the pars reticulata. Following quinolinic acid-induced degeneration of the striatonigral pathway, there was a marked loss of GABA immunoreactivity and a 59% increase in the density of GABAA receptors in the substantia nigra pars reticulata. There was a corresponding regional topography in the pattern of loss of GABA immunoreactivity and in the pattern of increase in GABAA receptors in the pars reticulata; the topography varied with the size and placement of the lesion in the striatum and correlated with the known topographical organization of the striatonigral projection. The quantitative autoradiographic results showed that following quinolinic acid lesions in the striatum: (i) the greatest increase in the density of GABAA receptors occurred in the middle third (91% increase) of the pars reticulata; (ii) the receptors were mainly of the GABAA/BZI variety; and (iii) 6-hydroxydopamine-induced degeneration of the dopaminergic pars compacta neurons did not significantly affect the density of receptors, indicating that the increased receptor binding was mainly localized on non-dopaminergic pars reticulata neurons. The immunohistochemical and in situ hybridization studies showed that, as in the normal substantia nigra, GABAA receptors in the substantia nigra pars reticulata on the lesioned side contained the alpha 1 and beta 2.3 GABAA receptor subtypes; the alpha 1 and beta 2.3 subtypes (but not the alpha 2) were increased after quinolinic acid lesions.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1994

Immediate early gene expression associated with the persistence of heterosynaptic long-term depression in the hippocampus.

Long-term depression (LTD) of synaptic efficacy is likely to be as important in memory processing as the more well-known long-term potentiation (LTP). The case for LTD serving as a memory mechanism, however, requires that it be shown to persist across days or weeks at least. Here we examined the persistence of heterosynaptic LTD in the medial and lateral perforant path inputs to the dentate gyrus in awake rats and correlated this persistence with the degree of immediate early gene expression as assessed immunohistochemically. Rats were chronically implanted with separate stimulating electrodes in the medial and lateral perforant paths and an extracellular field potential recording electrode in the dentate hilus. After recovery from surgery, either the medial or the lateral perforant path was tetanized with 400-Hz trains, and homosynaptic LTP and heterosynaptic LTD were followed across time. Heterosynaptic LTD was shown to occur readily in awake animals and to persist across days or weeks, depending on the stimulation protocol. The persistence of LTD and LTP was highly correlated within animals. Additional animals, given the same tetanization protocols, showed that the greatest immediate early gene expression occurred following that protocol which consistently gave the longest-lasting LTP and LTD. These data support the proposed role of LTD in memory processing but question whether immediate early genes are important for the persistence of LTP, LTD, or both.

Animals↗
PubMed1994

MK-801 and NBQX prevent electrically induced status epilepticus.

MUSCARINIC, NMDA and metabotropic glutamate receptor antagonists were tested for anticonvulsant effects in a continuous hippocampal stimulation model in rats in order to identify the receptors involved in the initiation of electrically induced status epilepticus (SE). Whereas the muscarinic receptor antagonists scopolamine and atropine and the metabotropic receptor antagonist L(+)-2-amino-3-phosphonopropionic acid (AP3) did not affect SE initiation, the N-methyl-D-aspartate (NMDA) antagonist dizocilpine (MK-801) (1 mg kg-1 i.p.) and the non-NMDA ionotropic receptor antagonist 2,3-dihydroxy-6-nitro-7-sulphamoylbenzo(F)-quinoxaline (NBQX) (two doses of 50 micrograms i.c.v.) prevented the induction of SE. It has been shown in a previous study that non-NMDA ionotropic receptors are involved in SE maintenance and it is now suggested that activation of NMDA receptors may principally initiate electrically induced SE, although non-NMDA ionotropic receptors may also be involved.

Animals↗
PubMed1994

Neuronal rescue with transforming growth factor-beta 1 after hypoxic-ischaemic brain injury.

Transforming growth factor-beta 1 (TGF-beta 1) mRNA is induced from 5 h to 3 days following hypoxic-ischaemic brain injury. Cell death also develops during this time suggesting that extracellular accumulation of this peptide may be involved in the processes that regulate cell loss. We examined the effect of rhTGF-beta 1 (0,2.5, 10,50 ng) injected into the cerebral lateral ventricle of rats 2 h after severe hypoxic-ischaemic brain injury. Histological outcome and B4-isolectin histochemistry were assessed 5 and 2 days, respectively following hypoxia. Treatment with 10 ng TGF-beta 1 reduced the microglia reaction (p < 0.05), the magnitude of neuronal loss (p < 0.01) and the area of cortical infarction (p < 0.05). Exogenous TGF-beta 1 given soon after hypoxic-ischaemic brain injury may have therapeutic potential and act by inhibiting the microglial reaction.

Animals↗
PubMed1994

MK-801 does not attenuate immediate-early gene expression following an amygdala afterdischarge.

It has been suggested that the increased transient expression of immediate-early gene transcription factors seen in nerve cells following an afterdischarge may initiate longer lasting or permanent changes in gene expression which underly the development of kindling. Since the development of kindling is sensitive to pharmacological blockade of the N-methyl-D-aspartate receptor, we tested whether the increased expression of the immediate-early genes c-fos, jun-B, c-jun, krox-20, and krox-24 following a kindling afterdischarge was also sensitive to N-methyl-D-aspartate receptor blockade by MK-801. In this report we demonstrate that all five immediate-early genes are induced by an amygdala afterdischarge. N-methyl-D-aspartate receptor blockade by a dose of MK-801 that significantly retards the development of amygdala kindling failed to attenuate immediate-early gene expression. These results suggest that although expression of these five immediate-early genes occurs after an amygdala afterdischarge their expression is not involved in the N-methyl-D-aspartate receptor-mediated component of amygdala kindling.

Amygdala↗
PubMed1994

c-fos antisense reduces expression of Krox 24 in rat caudate and neocortex.

1. The aim of this study was to investigate the neurochemical effects and measure the anatomical spread of infusion of c-fos antisense (AS) DNA into the striatum. 2. Rats were anesthetized and infused in opposing striata with c-fos AS and c-fos sense (S) DNA. Ten hours later they were injected with apomorphine (2 mg/kg, i.p.) and 20 min later they were overdosed with sodium pentobarbital and their brains either perfused or frozen. Vibratome-cut sections were immunostained for the detection of c-fos, JunB, Krox 24, somatostatin, substance P, dynorphin, tyrosine hydroxylase, and enkephalin. Cryostat-cut sections from the caudate were immunostained for the detection of c-fos, JunB, and Krox 24, as well as in situ hybridization for proenkephalin mRNA. Sections from the globus pallidus were used for the autoradiographic localization of D2 dopamine and A2a adenosine receptors. Sections from the substantia nigra were used for the autoradiographic localization of D1 dopamine and cannabinoid receptors. A second group of rats were injected in opposing striata with biotin-labeled c-fos AS DNA and c-fos S DNA. Ten hours later they were challenged with apomorphine (2 mg/kg, i.p.) and 20 min later brains were either perfused or frozen. Sections from these brains were cut throughout the rostral-caudal extent of the forebrain and the biotin labeled AS DNA was localized. 3. Krox 24 was expressed at high levels on the sense side of the brain in the striatum and overlying neocortex. However, on the AS-injected side there was a reduction in Krox 24 expression in striatum and overlying cortex. The biotin-labeled AS studies confirmed that the striatal infusion spread throughout the dorsal striatum as well as the overlying neocortex. We did not detect any changes in neurotransmitter receptors, neuropeptides, or tyrosine hydroxylase in AS/S-injected rat brains. 4. These results demonstrate that c-fos AS reduces Krox 24 expression in striatal and neocortical neurons but does not change the expression of a number of other proteins involved in basal ganglia function. Whether this effect is due to nonspecific actions of c-fos AS or to its effects on a component of the transduction pathway responsible for basal Krox 24 expression (NMDA receptors?) is unknown.

Animals↗