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Biomedical subjects

M Dragunow

Publications and source records attributed to M Dragunow.

At least 109 records · Page 6Linked to original sources

MK-801 sensitizes rats to pilocarpine induced limbic seizures and status epilepticus.

High doses of pilocarpine are known to produce prolonged seizure activity in rats. Previous studies show that pretreatment with LiCl will sensitize rats to the convulsant effects of pilocarpine. Here we report that the anti-epileptogenic and anticonvulsant drug MK-801 (dizocilpine) paradoxically also sensitizes rats to pilocarpine induced limbic seizures and limbic status epilipticus (LSE). If rats were pretreated with MK-801, normally subconvulsive doses of pilocarpine rapidly produced LSE. LSE could be prevented by co-administration of either atropine or diazepam with MK-801.

Animals↗

Correlation between the induction of an immediate early gene, zif/268, and long-term potentiation in the dentate gyrus.

Expression of the immediate early gene zif/268 (also termed NGFI-A, Krox 24, TIS8 and Egr-1) was investigated in awake rats following various long-term potentiation (LTP) induction protocols. zif/268 mRNA (Northern blots) and protein (immunohistochemistry) levels sharply increased following LTP, and followed a time course characteristic of other immediate early genes. When measured across 3 tetanization protocols known to produce differing degrees of LTP persistence, zif/268 induction was found to be more highly correlated with LTP duration than with the magnitude of initial LTP. These data support the hypothesis that the immediate early gene zif/268 plays a role as a third messenger in the cascade of cellular and nuclear events that govern the persistence of LTP.

Animals↗

A role for IGF-1 in the rescue of CNS neurons following hypoxic-ischemic injury.

Three days after unilateral hypoxic-ischemic injury in infant rats insulin-like growth factor 1 (IGF-1) production by astrocytes was enhanced in the injured region. This was associated with increased expression of mRNA for IGF binding protein-3 but not for binding protein-1. In adult rats a single lateral cerebroventricular injection of IGF-1 two hours following a similar injury markedly reduced neuronal loss. It is suggested that endogenous IGF-1 is neurotrophic and that centrally administered IGF-1 may have therapeutic potential for brain injury.

Animals↗

Induction of immediate-early gene proteins in dentate granule cells and somatostatin interneurons after hippocampal seizures.

The expression of the protein products of the immediate-early genes c-fos, Fos B, Fos-related proteins (FRAs), c-jun, jun B, jun D and krox-24 was investigated in the rat hippocampus at various times after electrically-induced hippocampal seizures. Hippocampal seizures induced all the immediate-early gene proteins in dentate granule cells with differing time-courses. In addition, Krox-24, Fos and Jun D were also induced in somatostatin-containing interneurons throughout the hippocampus and also in a small percentage of parvalbumin-containing interneurons. Thus, hippocampal seizures induce waves of immediate-early gene protein expression in dentate granule cells and a selective expression of krox-24, Fos and Jun D in hippocampal somatostatin interneurons. These results suggest that biochemical and/or morphological changes occurring in dentate granule cells and somatostatin interneurons after seizures may be regulated by immediate-early gene expression, and that these immediate-early gene proteins may be involved in seizure development in the nervous system.

Animals↗

Axotomized medial septal-diagonal band neurons express Jun-like immunoreactivity.

The expression of the transcription factors Fos and Jun was studied in rat brain after transection of the fornix-fimbria (FF) using polyclonal antibodies to these proteins and immunocytochemical detection methods. FF-transection lead to a massive induction of Jun-like immunoreactivity (JLI) in neurons in the medial septal nucleus and in the vertical limb of the diagonal band of Broca, within 48 hours and lasting up to 14 days after lesion. Fos was not induced in these neurons after FF-transection. These results indicate that axotomized medial septal and diagonal band of Broca neurons selectively and rapidly express JLI. The role of Jun expression in axonal regeneration or neuronal death is discussed.

Acetylcholinesterase↗

Accumulation of calcitonin-gene related peptide-like immunoreactivity after hypoxic-ischaemic brain injury in the infant rat.

Unilateral carotid ligation in immature rats, followed by either 15 min (moderate group) or 90 min (severe group) of hypoxia were used to assess the effects of hypoxia-ischaemia (HI) on the accumulation of the neuropeptide calcitonin-gene related peptide (CGRP). Severe, but not moderate, HI produced a massive time-dependent increase in CGRP-like immunoreactivity throughout the damaged regions of the brain (neocortex, caudate-putamen, hippocampus) beginning at 24 h and maximal at 3-5 days after HI. By 11 days after HI levels appeared to have returned to baseline. The increased immunoreactivity was largely localized to presumed axon terminals contacting neurons, blood vessels and non-nerve cells. Scattered neurons in the cingulate cortex, piriform cortex and striatum also showed increased immunoreactivity in their soma. These results raise the possibility that CGRP may be involved in neuronal repair after HI in the infant brain.

Animals↗

Basal expression of Fos, Fos-related, Jun, and Krox 24 proteins in rat hippocampus.

The basal expression of the protein products of the inducible immediate early genes (IEGs), Fos, Jun, and Krox 24, was investigated in rat hippocampus using immunocytochemical visualization methods with antisera specific for Fos only, Fos and the Fos-related antigens (FRAs), the Jun family, and Krox 24 (previously described as TIS 8, egr-1, NGF-IA or zif 268). In the normal adult rat brain basal levels of Jun, Krox 24 and Fos-related antigens but not Fos were seen within the hippocampus. More specifically very high basal levels of Jun were seen in the dentate granule cells with high basal Krox 24 levels seen in the CA1-subiculum region of the rat hippocampus. Basal FRAs but not Fos-positive cells were seen at low levels in the dentate granule cells. The implications of these results to the functioning of IEG proteins in hippocampal neurons is discussed.

Animals↗

The regional, cellular and subcellular localization of GABAA/benzodiazepine receptors in the substantia nigra of the rat.

The regional, cellular and subcellular distribution of GABAA/benzodiazepine receptors was investigated by light and electron microscopy in the rat substantia nigra. The regional distribution and density of GABAA/benzodiazepine receptor subtypes (Type I and II) was studied using quantitative receptor autoradiography following in vitro labelling of cryostat sections with tritiated ligands. This was followed by a detailed study of the cellular and subcellular distribution and localization of GABAA/benzodiazepine receptors by light and electron microscopy using immunohistochemical techniques with a monoclonal antibody (bd-17) to the beta 2,3 subunits of the GABAA/benzodiazepine receptor complex. Finally, in situ hybridization histochemistry using 35S-labelled oligonucleotide probes was used to demonstrate the cellular distribution of mRNA for the alpha 1 and alpha 2 GABAA receptor subunits in the substantia nigra. The results of the autoradiographic and immunohistochemical studies showed a close correspondence in the regional distribution of GABAA/benzodiazepine receptors in the substantia nigra. A moderate-to-high density of receptors was present throughout the full extent of the substantia nigra pars reticulata with a very low density of receptors in the substantia nigra pars compacta. Quantitative autoradiographic studies showed that: (i) the pars reticulata contained mainly central Type I receptors; (ii) the highest density of receptors was present in the caudal pars reticulata (200 +/- 38 fmol/mg) with successively lower densities of receptors in the middle (176 +/- 31 fmol/mg) and rostral (150 +/- 26 fmol/mg) levels of the pars reticulata; and (iii) the density of receptors in the pars reticulata was reduced by 34% following 6-hydroxydopamine-induced degeneration of dopaminergic pars compacta neurons. At the cellular level, GABAA/benzodiazepine receptor immunoreactivity was localized in a punctate fashion on dendrites and neuronal cell bodies in the pars reticulata. At the subcellular level, GABAA/benzodiazepine receptor immunoreactivity was associated with the pre- and postsynaptic membranes of axodendritic synaptic complexes along the length of small-to-large sized smooth dendrites in the pars reticulata. Two types of immunoreactive axodendritic synaptic complexes were identified: most (about 80%) immunopositive synapses showed equal staining of the pre- and postsynaptic membranes and were associated with small (less than 1.0 micron) axon terminals containing few mitochondria and small, round-to-pleomorphic vesicles in synaptic contact with small, peripheral dendrites; less frequently (about 20%) immunopositive synapses showed a marked immunoreactive thickening of the postsynaptic membrane and were associated with large (greater than 1.0 micron) axon terminals containing numerous mitochondria and mainly pleomorphic vesicles in synaptic contact with large mainstem dendrites.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Distribution of excitatory and inhibitory amino acid, sigma, monoamine, catecholamine, acetylcholine, opioid, neurotensin, substance P, adenosine and neuropeptide Y receptors in human motor and somatosensory cortex.

Autoradiography was used to visualise N-methyl-D-aspartate, phencyclidine, strychnine-insensitive glycine, alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid, kainic acid, benzodiazepine, gamma-aminobutyric acid type A, sigma, serotonergic, dopaminergic, alpha 2-adrenergic, beta-adrenergic, muscarinic cholinergic, nicotinic, opioid, neurotensin, substance P, adenosine A1 and neuropeptide Y receptors in the human primary motor (Brodmann's area 4) and somatosensory cortex (Brodmann's areas 3, 2 and 1). With the exception of serotonin type 2 receptors, all receptor types examined had a similar distribution in area 4 which showed little dependence on the underlying distribution of cell somata, often continuing unaltered through the somatosensory cortex despite marked cytoarchitectural changes. The highest densities occurred in the outer (most superficial) 30-40% of the cortical grey matter, followed by a band of relatively low binding and then moderate levels in the inner (deeper) region. In many instances, an additional band of dense binding could be discerned in the region of laminae IV/Va running unbroken through both gyri. The distribution of most receptor types in the somatosensory cortex also followed this pattern, except for opioid and kainic acid receptors which showed higher levels in the inner rather than the outer third of this region. At the edge of area 4, a change occurred such that a high density outer band appeared, giving these receptor types the same pattern in area 4 as the majority. Serotonin type 2 receptor levels were quite low in the outermost region of area 4, although the pattern was otherwise similar to that of the other receptors. Thus, with the exception of serotonin receptors, the similarity in many binding site distributions recently noted in area 4 of the rhesus monkey also tends to occur in the human area 4, to the extent that 2 ligands will reverse their usual cortical binding pattern to conform with the common area 4 pattern.

Autoradiography↗

Autoradiographic distribution of sigma receptors in human neocortex, hippocampus, basal ganglia, cerebellum, pineal and pituitary glands.

Autoradiographic distributions of [3H]1,3-di-O-tolylguanidine, [3H]DTG, binding to sigma (sigma) receptors were studied in key human brain regions. High densities occurred in the substantia nigra pars compacta and cerebellum. Pineal and pituitary levels were moderate. In neocortex, binding was high in laminae II-IVA and much lower in the midzone. Moderate binding occurred over hippocampal dentate granular cells and in the striatum. These results differ from those reported for the rodent.

Adult↗

Elevated expression of jun and fos-related proteins in transplanted striatal neurons.

The basal expression of the immediate-early gene protein products fos, fos-related antigens (FRA's), jun and krox-24 was examined using immunocytochemical methods in intrastriatal grafts derived from fetal striatal primordia. Whereas very few, if any, normal adult striatal neurons expressed jun, many neurons in the striatal graft expressed jun at high levels. FRA's, but not fos, were also occasionally induced in some grafted striata. krox-24 was expressed in normal adult striatal neurons, and to a lesser extent in transplanted striatal neurons. These results show that neurons of intrastriatal grafts express jun at substantially higher levels than host striatal neurons, and this may be related to the previously reported increased transcription of neuropeptides in striatal grafts, and/or to the possible failure of transplanted neurons to fully establish normal connections with the host tissue.

Animals↗

Autoradiographic visualisation of [3H]DTG binding to sigma receptors, [3H]TCP binding sites, and L-[3H]glutamate binding to NMDA receptors in human cerebellum.

The autoradiographic distributions of [3H]1,3-di-ortho-tolyguanidine ([3H]DTG), [3H]1-[1-(2-thienyl) cyclohexyl] piperidine ([3H]TCP) and L-[3H]glutamate were studied in the human cerebellum. [3H]DTG is a selective label for the sigma receptor, while L-[3H]glutamate binding was carried out under conditions selective for the N-methyl-D-aspartate (NMDA) receptor. [3H]TCP binding sites and sigma receptors showed marked enrichment in the Purkinje cell layer, while L-[3H]glutamate-labelled NMDA receptors showed virtually no binding in the Purkinje cell layer. The results confirm the existence of [3H]TCP binding sites which are not linked to NMDA receptors in the human cerebellum, having a distribution which is more similar to that of the haloperidol-sensitive sigma receptor.

Autoradiography↗

Differential sensitivity of calbindin and parvalbumin immunoreactive cells in the striatum to excitotoxins.

The neurotoxic effects of ibotenic acid, quinolinic acid and kainic acid on cells in the rat striatum were investigated using immunocytochemistry with antibodies to the calcium binding proteins, calbindin and parvalbumin. The results showed that both ibotenic acid and quinolinic acid affected calbindin and parvalbumin cells to the same extent. However, parvalbumin immunopositive neurons were more sensitive than calbindin immunopositive neurons to the neurotoxic effects of kainic acid. Although the reason for this increased sensitivity of parvalbumin striatal neurons to kainic acid is unclear, these results suggest that the neurotoxicity produced by kainic acid is different to that occurring with quinolinic acid and ibotenic acid.

Animals↗

3,4-Methylenedioxymethamphetamine induces Fos-like proteins in rat basal ganglia: reversal with MK 801.

Injections of 3,4-methylenedioxymethamphetamine (MDMA, 25 mg/kg, i.p.) to rats lead to an accumulation of c-fos protein (Fos) and Fos-related antigens in caudate-putamen, nucleus accumbens and olfactory tubercle. This induction occurred at least 2 h (but not at 10 min) after injection and Fos levels had returned to baseline after 24 h, although Fos-related antigens remained elevated 24 h after injection. The NMDA antagonist MK 801 inhibited Fos and Fos-related antigen induction after MDMA injections, whereas fluoxetine, a serotonin uptake inhibitor, had no effect. Thus, MDMA induces Fos and Fos-related antigens in striatal neurons in an NMDA-reversible fashion.

3,4-Methylenedioxyamphetamine↗

The role of immediate early genes in the stabilization of long-term potentiation.

Immediate early genes (IEGs) are a class of genes that show rapid and transient but protein synthesis-independent increases in expression to extracellular signals such as growth factors and neurotransmitters. Many IEGs code for transcription factors that have been suggested to govern the growth and differentiation of many cell types by regulating the expression of other genes. IEGs are expressed in adult neurons both constitutively and in response to afferent activity, and it has been suggested that during learning, IEGs may play a role in the signal cascade, resulting in the expression of genes critical for the consolidation of long-term memory. Long-term potentiation (LTP) is a persistent, activity-dependent form of synaptic plasticity that stands as a good candidate for the mechanism of associative memory. A number of IEGs coding for transcription factors have been shown to transiently increase transcription in the dentate gyrus of rats following LTP-inducing afferent stimulation. These include zif/268 (also termed NGFI-A, Krox-24, TIS-8, and egr-l), c-fos-related genes, c-jun, junB, and junD. Of these, zif/268 appears to be the most specifically related to LTP since it is evoked under virtually all LTP-inducing situations and shows a remarkably high correlation with the duration of LTP. There are a number of outstanding questions regarding the role of zif/268 and other IEGs in LTP, including which second messenger systems are important for activating them, which "late effector" genes are regulated by them, and the exact role these genes play, if any, in the stabilization and maintenance of LTP.

Animals↗

Prolonged and selective induction of Fos-related antigen(s) in striatal neurons after 6-hydroxydopamine lesions of the rat substantia nigra pars compacta.

Unilateral 6-hydroxydopamine (6-OHDA) lesions of the rat substantia nigra lead to a large widespread and long-lasting (greater than 3 months) increased expression of Fos-related antigen(s) (FRAs) in striatal neurons ipsilateral to the side of the lesion. However, Fos and Jun expression were only very slightly increased in a few scattered neurons in the dopamine-denervated striatum. These results demonstrate that FRAs are induced long-term in striatal neurons following dopamine-depletion. This increased production of FRAs may be related to neuropeptide and/or D2 dopamine receptor upregulation that also occurs in the dopamine-denervated striatum.

Animals↗

Presence and induction of Fos B-like immunoreactivity in neural, but not non-neural, cells in adult rat brain.

The presence and induction of Fos B, a novel growth factor-activated gene, was investigated in adult rat brain using an antiserum to Fos B and immunocytochemical methods. In normal rat brain immunoreactivity was detected in the nuclei of nerve cells scattered in the cerebral cortex, striatum, amygdala, hippocampus and dentate gyrus. This immunostaining was not present in adjacent brain sections incubated with anti-Fos B serum preadsorbed with the Fos B peptide. Furthermore, Fos B-like immunoreactivity was induced in neurons by two treatments (focal brain injury and haloperidol injection) that are known to induce Fos, however, whereas Fos levels returned to baseline 24 h after these treatments, Fos B-like immunoreactivity remained elevated at this time point. Also, although focal brain injury and rolipram injections induced Fos in ependymal and glial-like cells in rat brain, Fos B-like immunoreactivity was not detected in these non-neural cells. The implications of these results for the presence of Fos B in adult neurons is discussed.

Animals↗