Benzoxazinones as potent positive allosteric AMPA receptor modulators: part I.

AMPA receptors (AMPARs) are an increasingly important therapeutic target in the CNS. Aniracetam, the first identified potentiator of AMPARs, led to the rigid and more potent CX614. This lead molecule was optimized in order to increase affinity towards the AMPA receptor.

Substituted benzoxazinones as potent positive allosteric AMPA receptor modulators: part II.

AMPA receptors (AMPARs) are an important therapeutic target in the CNS. A series of substituted benzoxazinone derivatives with good to very good in vitro activity as positive allosteric AMPAR modulators was synthesized and evaluated. The appropriate substituent choice on the benzoxazinone fragment improved the affinity towards the AMPA receptor significantly in comparison to our lead molecule CX614.

Expression of metabotropic glutamate receptor 5 is increased in astrocytes after kainate-induced epileptic seizures.

In the CNS there is a differential distribution of the metabotropic glutamate receptor 5 (mGluR5) in neurons and glia. Hippocampal nerve cells contain large amounts of the receptor transcript and protein that are expressed at very low levels in astrocytes. This is unexpected, as mGluR-induced phosphoinositide hydrolysis is substantial in cultured astrocytes and is mediated only by mGluR5 in these cells.

Postnatal growth of corticospinal axons in the spinal cord of developing mice.

The corticospinal tract (CST) plays an important role in the control of voluntary movements. Although the development of the CST has been studied extensively in other species, limited information is available on its development in mice. In the present study, the growth of corticospinal axons was characterized in developing mice using Phaseolus vulgaris leucoagglutinin (PHA-L).

Decreased expression of N-methyl-D-aspartate receptor 1 messenger RNA in select regions of Alzheimer brain.

An antisense oligonucleotide probe was used to examine the expression of gene encoding the obligatory NMDAR1 subunit of the N-methyl-D-aspartate receptor in the hippocampus and adjacent cortical areas (entorhinal and perirhinal cortices) of seven Alzheimer patients and in the same brain regions of seven control individuals. Both groups were matched according to age, sex, cause of death, post mortem delay, and tissue storage time. Densitometric analysis of in situ hybridization autoradiograms revealed a 34% (P<0.

Dopaminergic denervation of striatum results in elevated expression of NR2A subunit.

An imbalance between glutamate and dopamine in the striatum may contribute to the pathophysiology of Parkinson's disease. We therefore studied the effect of dopaminergic denervation of the rat striatum (unilateral 6-OHDA lesions of the medial forebrain bundle) on NMDA receptors. The expression of NMDA receptor genes (NR1, NR2A-B) was examined by in situ hybridization using oligonucleotide probes, and binding to NMDA receptors assessed using L-[3H]glutamate.

Selective increase of NMDA-sensitive glutamate binding in the striatum of Parkinson's disease, Alzheimer's disease, and mixed Parkinson's disease/Alzheimer's disease patients: an autoradiographic study.

Parkinson's disease (PD) and Alzheimer's disease (AD) may share certain abnormalities since a subset of PD patients suffer from dementia, and some AD individuals show extrapyramidal symptoms. In vitro quantitative autoradiography was used to examine different subtypes of excitatory amino acid (EAA) receptors (NMDA, KA, and AMPA) and dopamine transporter sites in the striatum (caudate, putamen) and nucleus accumbens (NAc) from idiopathic PD, pure AD, and mixed PD/AD patients. PD and AD groups, and to a lesser extent the PD/AD groups, showed substantially increased binding to NMDA receptors in the striatum and NAc.

Chronic haloperidol treatment enhances binding to NMDA receptors in rat cortex.

Hyperactivity of the dopaminergic system and a hypoglutamatergic state have been hypothesized to underlie schizophrenia. It has also been proposed that neuroleptics may interact not only with the dopaminergic system but also with the glutamatergic system. We found that daily intraperitoneal injections of haloperidol (1 mg kg-1) for 21 days resulted in increased binding (10-20%) to the NMDA type of glutamate receptors in the outer layers of rat parietal cortex.

Increased sensitivity to adenosine in the rat dentate gyrus molecular layer two weeks after partial entorhinal lesions.

The molecular layer of the dentate gyrus exhibits extensive circuit and receptor reorganization after entorhinal lesions and in Alzheimer's disease, including decreased adenosine (A1) receptor binding in the terminal zone of damaged perforant path fibers. We examined the adenosine-sensitivity of evoked synaptic activity recorded from the rat dentate gyrus molecular layer in hippocampal slices prepared after electrolytic lesions were placed in approximately the middle third of the entorhinal cortex. Extracellular field potentials (EFPs) recorded in slices prepared from animals two days post-lesion were small, upward-going, and exhibited paired-pulse potentiation, but by two weeks post-lesion EFPs had recovered to large, downward-going responses that exhibited paired-pulsed depression.

Reduced density of adenosine A1 receptors and preserved coupling of adenosine A1 receptors to G proteins in Alzheimer hippocampus: a quantitative autoradiographic study.

Binding to adenosine A1 receptors and the status of their coupling to G proteins were studied in the hippocampus and parahippocampal gyrus of Alzheimer individuals and age-matched controls. The binding to A1 receptors was compared with binding to the N-methyl-D-aspartate receptor complex channel-associated sites (labeled with (+)-[3H]5-methyl-10,11-dihydro-5H- dibenzo[a,d]cyclohepten-5,10-imine maleate). In vitro quantitative autoradiography demonstrated a similar anatomical distribution of A1 receptors labeled either with an agonist ((-)-[3H]phenylisopropyladenosine) or antagonist ([3H]8-cyclopentyl-1,3-dipropylxanthine) in the brains of elderly controls.

Excitatory amino acid receptors in schizophrenia.

Growing evidence suggests an involvement of excitatory amino acid (EAA) systems in schizophrenia. Precedent exists for changes in binding to kainate, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid, N-methyl-D-aspartate subtypes of EAA receptors. Current evidence indicates that in schizophrenia, EAA receptor levels can be decreased, unchanged, or even increased in certain brain regions and certain cases.

Elevated NMDA receptors in parkinsonian striatum.

Dopamine-glutamate interactions contribute to normal striatal function and have been implicated in neurotoxicity at nigrostriatal dopamine (DA) terminals. The present study examined the striata of idiopathic Parkinson's disease (PD) patients and age-matched controls for regional differences in the DA transporter and binding to N-methyl-D-aspartate (NMDA) receptors. [3H]Mazindol labeling of the DA transporter was reduced by 70-80% in the caudate and putamen of PD patients, with reductions being more extensive dorsally than ventrally.

Hippocampal excitatory amino acid receptors in elderly, normal individuals and those with Alzheimer's disease: non-N-methyl-D-aspartate receptors.

Quantitative receptor autoradiography was used to examine the density and distribution of [3H]kainic acid and [3H]alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) binding sites in the hippocampal formation and parahippocampal gyrus obtained at autopsy from 10 Alzheimer's disease and eight normal control individuals. In control and Alzheimer's disease individuals, [3H]kainic acid saturation binding analysis in the outer molecular layer of the dentate gyrus fitted a single-site model. Added calcium ions did not alter the density of [3H]kainic acid binding in the human tissues.

N-methyl-D-aspartate receptor complex in the hippocampus of elderly, normal individuals and those with Alzheimer's disease.

The various ligand binding sites of the N-methyl-D-aspartate receptor complex in the hippocampal formation and parahippocampal gyrus of Alzheimer's disease patients and age-matched normal individuals were examined using quantitative autoradiography. The hippocampus and parahippocampal gyrus of the normal elderly brain exhibited virtually identical distributions of L-[3H]glutamate, [3H]5-methyl-10,11-dihydro-5H- dibenzo[a,d]cyclohepten-5,10-iminemaleate ([3H]MK-801), [3H][(+/-)2-carboxypiperazine-4-yl]propyl-1-phosphonic acid ([3H]CPP) and strychnine-insensitive [3H]glycine binding sites (r greater than 0.87) suggesting that binding occurred to different domains of the same receptor macromolecule.

Kainate receptors in the rat hippocampus: a distribution and time course of changes in response to unilateral lesions of the entorhinal cortex.

The response of kainate receptors to deafferentation and subsequent reinnervation following unilateral entorhinal cortex lesions was studied in the rat hippocampus using quantitative in vitro autoradiography. The binding levels of [3H]kainic acid (KA) and changes in the distribution of KA sites were investigated in the dentate gyrus molecular layer (ML) and in various terminal zones in the CA1 field at 1, 3, 7, 14, 21, 30, and 60 d postlesion. The data from both the ipsilateral and contralateral hippocampus were compared with those from unoperated controls.

Plastic response of hippocampal excitatory amino acid receptors to deafferentation and reinnervation.

In vitro autoradiography was used to examine the response of excitatory amino acid receptors in the hippocampus of rat following unilateral lesions of the entorhinal cortex. The density of N-methyl-D-aspartate and quisqualate receptor binding was determined on days 1, 3, 7, 14, 21, 30 and 60 postlesion both ipsilateral and contralateral to the lesion and in unoperated controls. The results are compared to the time-course of deafferentation and reinnervation.

Muscarinic acetylcholine receptors in bulk-isolated astrocytes of the hippocampus in the rat.

The high-affinity muscarinic antagonist L-[3H]quinuclidinyl benzi-late (L-[3H]QNB) was used to label muscarinic acetylcholine receptors in the astrocyte-enriched fraction isolated from rat hippocampus. Analysis of the saturation and drug inhibition data showed that [3H]QNB binding in the astroglia fraction was saturable, of high affinity (Kd = 0.026 nM) and competitively inhibited by muscarinic antagonists and agonists.

Differential response of hippocampal muscarinic cholinergic receptors to various deafferentations of the hippocampus in the rat.

The influence of bilateral electrolytic lesions of different parts of the septum on muscarinic receptor binding in the hippocampus was studied within 14-21 days after operation. The effect of total septal lesion upon receptor binding was also investigated separately in the dorsal and ventral hippocampus and in five consecutive hippocampal parts along the septotemporal axis of the structure. The data indicate that: (i) differential response of muscarinic receptors, as revealed by a decrease, increase or lack of changes in the [3H]QNB binding depends on the site and extent of the lesion, (ii) lack of changes in muscarinic receptor binding can be spurious when the investigation is performed on the whole hippocampus, and masked by regional response differences, (iii) differential response of [3H]QNB binding sites in distinct parts of the hippocampus to total septal lesion may depend on the preexisting differences in the density of cholinergic innervation and of muscarinic receptors.

Analysis of the time course of GM1 ganglioside effect on changes in choline acetyltransferase activity in partially denervated rat hippocampus.

The effect of chronic GMl ganglioside administration (30 mg/kg, daily) for 6, 21, 42 and 90 days on the activity of choline acetyltransferase was investigated in the hippocampus of rats with partial electrolytic lesions of the dorsal hippocampal afferents and in unoperated rats. No influence of GM1 administration on ChAT activity was noted in unoperated animals. The lesions caused denervation in the hippocampus, which occurred with varying intensity along its dorsoventral axis, as shown by the gradual pattern of decrease in ChAT activity.

Effect of GM1 ganglioside treatment on postlesion responses of cholinergic enzymes in rat hippocampus after various partial deafferentations.

The effect of intramuscular administration of monosialoganglioside (GM1) on postlesion responses of choline acetyltransferase and acetylcholinesterase activity in partially deafferented rat hippocampus was studied at various survival times. Lesions partially destroying the medioventral, septal area, or lesions performed in supracallosal stria including corpus callosum and cingulum evoked cholinergic denervation of the hippocampus, while those made in entorhinal cortex resulted in partial glutamatergic deafferentation. GM1 treatment potentiates the responses of both cholinergic enzymes, independently of whether the partial deafferentation was homo- or heterotypical.

The stimulating effect of ganglioside injections on the recovery of choline acetyltransferase and acetylcholinesterase activities in the hippocampus of the rat after septal lesions.

The effect of intramuscular administration of a mixture of gangliosides (21% GM1, 39.7% GD1a, 16% GD1b, 19% GT1) in a daily of 50 mg per kg upon the time course of changes in hippocampal acetylcholinesterase and choline acetyltransferase activities after extensive medioventral septal lesions in the rat was checked on days 3, 5, 18 and 50 after the operation. Following the early decrease in the enzyme activities to about 25% of control due to degeneration, a gradual recovery up to about 50% of control activity at the 50th day was found.