Efficacy of rosiglitazone in a genetically defined population with mild-to-moderate Alzheimer's disease.

Mild-to-moderate AD patients were randomized to placebo or rosiglitazone (RSG) 2, 4 or 8 mg. Primary end points at Week 24 were mean change from baseline in AD Assessment Scale-Cognitive (ADAS-Cog) and Clinician's Interview-Based Impression of Change Plus Caregiver Input global scores in the intention-to-treat population (N=511), and results were also stratified by apolipoprotein E (APOE) genotype (n=323). No statistically significant differences on primary end points were detected between placebo and any RSG dose. There was a significant interaction between APOE epsilon4 allele status and ADAS-Cog (P=0.014). Exploratory analyses demonstrated significant improvement in ADAS-Cog in APOE epsilon4-negative patients on 8 mg RSG (P=0.024; not corrected for multiplicity). APOE epsilon4-positive patients did not show improvement and showed a decline at the lowest RSG dose (P=0.012; not corrected for multiplicity). Exploratory analyses suggested that APOE epsilon4 non-carriers exhibited cognitive and functional improvement in response to RSG, whereas APOE epsilon4 allele carriers showed no improvement and some decline was noted. These preliminary findings require confirmation in appropriate clinical studies.

Increased cyclic AMP reduces 5-HT1D receptor-mediated inhibition of [3H]5-hydroxytryptamine release from guinea-pig cortical slices.

This study investigated the role of cyclic AMP in the inhibition of [3H]5-hydroxytryptamine ([3H]5-HT) release mediated through the 5-HT1D autoreceptor using slices of guinea-pig cerebral cortex. Forskolin, dibutyryl-cyclic AMP and rolipram, which increase intracellular cyclic AMP by different mechanisms, all attenuated the inhibitory effect of the autoreceptor agonist 5-carboxamidotryptamine. These results indicate that modulation of cyclic AMP production affects 5-HT1D autoreceptor function.

Inhibition of excitatory amino acid-stimulated phosphoinositide hydrolysis in rat hippocampus by L-aspartate-beta-hydroxamate.

The effect of a series of glutamate uptake inhibitors was tested on ibotenate-stimulated phosphoinositide hydrolysis. The pharmacological profile of the inhibitory effect of these compounds on the ibotenate response was quite different from that on glutamate uptake. Aspartate-beta-hydroxamate was the most potent compound with the L-isomer (IC50 11 +/- 2 microM) being considerably more potent than the D-isomer (IC50 104 +/- 12 microM). The effect of the L-aspartate-beta-hydroxamate was found to be specific for ibotenate and quisqualate-stimulated phosphoinositide hydrolysis; this compound did not affect hydrolysis stimulated by carbachol, K+ or sodium fluoride. The inhibition of the ibotenate response was found to involve a non-competitive and irreversible mechanism.

Seizures selectively impair agonist-stimulated phosphoinositide hydrolysis without affecting protein kinase C activity in rat brain.

The influence of seizures on phosphoinositide hydrolysis and protein kinase C activity was measured in rat hippocampus and cerebral cortex, primarily using a model in which generalized convulsive status epilepticus was induced by administration of LiCl (3 mmole/kg) 20 hr prior to pilocarpine (30 mg/kg). A short (5 min) period of seizures reduced phosphoinositide hydrolysis in hippocampal slices stimulated by norepinephrine or ibotenate, but did not alter the responses to carbachol, 50 mM K+, or NaF. Induction of seizures with diisopropylfluorophosphate caused a similar reduction in the response to norepinephrine without altering carbachol-stimulated phosphoinositide hydrolysis. The inhibition of norepinephrine-stimulated phosphoinositide hydrolysis after seizures generated by lithium plus pilocarpine administration was apparently not due to inhibitory influences of quisqualate or activation of protein kinase C since both of these treatments caused similar inhibitions in slices from control and treated rats. Seizures induced by lithium plus pilocarpine or by kainate did not alter the activity of protein kinase C or the distribution of protein kinase C between membrane and cytosolic fractions. Thus, seizures cause a neurotransmitter-selective impairment of phosphoinositide hydrolysis, and this response may play a role in the severity or duration of seizure activity.

Pertussis toxin potentiates seizures induced by pilocarpine, kainic acid and N-methyl-D-aspartate.

Previous studies with lithium have shown that it potentiated the in vivo response to cholinomimetics in rats, resulting in seizures at otherwise non-convulsant doses, but did not affect seizure activity induced by a number of chemical convulsants including kainic acid and N-methyl-D-aspartate (NMDA). In vitro experiments have suggested that lithium interferes with receptor-mediated second messenger production, possibly due to an action at G-proteins. The present study tested the hypothesis that selective inhibition of G-proteins by in vivo administration of pertussis toxin would induce effects similar to those of lithium. The results reported here demonstrate that pertussis toxin mimics lithium in potentiating the convulsant response to pilocarpine in rats. The effect of pertussis toxin was dose-dependent and the extent of potentiation was over 13-fold, which was remarkably similar to lithium. The seizures were prevented by pretreatment with atropine, phenobarbital or diazepam. L-Phenylisopropyladenosine (L-PIA) and MK-801 also demonstrated anticonvulsant activity, with MK-801 also protecting the rats against the rapid death associated with pertussis toxin/pilocarpine-induced seizures. Thus, seizures were cholinergically initiated and were controlled by the same drugs as were lithium/pilocarpine-induced seizures. The results illustrate that in several respects the response to cholinomimetics is modified in a similar manner by lithium and pertussis toxin. However, pertussis toxin lacks the specificity of lithium as it also potentiated the convulsant effects of kainic acid and NMDA.

Analysis of the convulsant-potentiating effects of lithium in rats.

Lithium pretreatment of rats has previously been shown to potentiate the convulsant effects of cholinomimetic drugs, such as pilocarpine. The first objective of this project was to determine if lithium also potentiates seizures induced by other classes of drugs. Lithium pretreatment of rats did not affect seizure activity induced by administration of N-methyl-D-aspartate, kainic acid, bicuculline, or pentylenetetrazole. This suggests that the proconvulsant effect of lithium is largely selective for cholinomimetics. A second series of experiments investigated possible mechanisms of the lithium potentiation of pilocarpine-induced seizures. The alpha 2-adrenergic receptor agonist clonidine suppressed seizure development, and the antagonist idazoxan enhanced the onset of seizures, suggesting that endogenous norepinephrine provides anticonvulsant properties. Administration of the norepinephrine depleter DSP-4 potentiated pilocarpine-induced seizures. These results suggest that the previously reported impairment of noradrenergic function by lithium may play a role in its potentiation of cholinomimetic-induced seizures.

Reduction of Na+ enhances phosphoinositide hydrolysis and differentiates the stimulatory and inhibitory responses to quisqualate in rat brain slices.

The concentration of Na+ in the incubation medium significantly influenced phosphoinositide hydrolysis induced by some, but not all, agonists in rat cerebral cortical slices. Reductions of the Na+ concentration below 120 mM resulted in incremental increases in basal and norepinephrine-stimulated accumulation of [3H]inositol monophosphate in cortical slices that had been prelabelled with [3H]inositol, and maximal responses were obtained with 0 and 5 mM Na+. In contrast, the responses to carbachol and ibotenate were similar in medium containing 120 or 5 mM Na+. In medium with 120 mM Na+, quisqualate has two effects on phosphoinositide hydrolysis in cortical slices, including a relatively weak stimulatory effect and an inhibitory modulation of the stimulation induced by norepinephrine. These two responses to quisqualate were differentially modulated by Na+; in 5 mM compared with 120 mM Na+ the stimulatory response was greatly increased and the inhibitory effect was mostly eliminated. That these were two separate events was confirmed by the use of L-BOAA (beta-N-oxalyl-L-alpha, beta-diaminopropionic acid), which reproduces the inhibitory, but not the stimulatory effect of quisqualate on phosphoinositide hydrolysis. In 5 mM Na+, inhibition by L-BOAA of norepinephrine-stimulated phosphoinositide hydrolysis was completely eliminated. These results demonstrate that a physiological concentration of Na+ maintains phosphoinositide hydrolysis at a submaximal level of sensitivity to some, but not all, agonists. The differential effects of Na+ on the stimulatory and inhibitory effects of quisqualate further substantiate the suggestion that these are two separate processes and indicate that alterations of the Na+ concentration may influence the effects of quisqualate, and other agonists, on phosphoinositide hydrolysis.

Dibenzocycloalkenimine (MK-801) stimulates phosphoinositide hydrolysis in rat cerebral cortical slices).

Dibenzocycloalkenimine (MK-801), a non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist, induced a concentration-dependent accumulation of inositol monophosphate in rat cerebral cortical slices. This effect appears unrelated to antagonism of NMDA receptors as AP-7 and PCP did not cause comparable effects. In addition, the MK-801-induced response was unaffected by NMDA or glycine, alone or in combination, Zn2+, or the removal of Mg2+ from the buffer. These results indicate a novel site of action for MK-801 associated with activation of phosphoinositide hydrolysis.

Inhibition of phosphoinositide hydrolysis by the novel neurotoxin beta-N-oxalyl-L-alpha, beta-diaminopropionic acid (L-BOAA).

Inhibition by a recently isolated neurotoxic amino acid, beta-N-oxalyl-L-alpha, beta-diaminopropionic acid, (L-BOAA), of stimulated phosphoinositide hydrolysis was studied in rat brain cerebral cortical slices. L-BOAA inhibited the norepinephrine-stimulated response but did not affect hydrolysis induced by 55 mM K+, carbachol, or carbachol in the presence of 20 mM K+. The inhibition was concentration-dependent with an IC50 of 300 microM. This inhibition was insensitive to the excitatory amino acid antagonists, gamma-glutamylglycine, glutamic acid diethyl ether, CNQX, AP-4, AP-7, or kynurenate. Thus, we propose that the L-BOAA-mediated inhibition of the norepinephrine-stimulated response was due to an interaction at a novel site, which may also be sensitive to quisqualate (see discussion). The mechanism of the inhibition is still unknown but was not prevented by inhibition of phospholipase A2 or polyamine synthesis and it was not affected by blockade of chloride channels. However, the presence of 20 mM K+ completely blocked the inhibitory effect of L-BOAA on norepinephrine-stimulated phosphoinositide hydrolysis.

Anticonvulsant actions of MK-801 on the lithium-pilocarpine model of status epilepticus in rats.

MK-801, a noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist, was tested for anticonvulsant effects in rats using two seizure models, coadministration of lithium and pilocarpine and administration of a high dose of pilocarpine alone. Three major results are reported. First, pretreatment with MK-801 produced an effective and dose-dependent anticonvulsant action with the lithium-pilocarpine model but not with rats treated with pilocarpine alone, suggesting that different biochemical mechanisms control seizures in these two models. Second, the anticonvulsant effect of MK-801 in the lithium-pilocarpine model only occurred after initial periods of seizure activity. This observation is suggested to be an in vivo demonstration of the conclusion derived from in vitro experiments that MK-801 binding requires agonist-induced opening of the channel sites of the NMDA receptor. Third, although it is relatively easy to block seizures induced by lithium and pilocarpine by administration of anticonvulsants prior to pilocarpine, it is more difficult to terminate ongoing status epilepticus and block the lethality of the seizures. Administration of MK-801 30 or 60 min after pilocarpine, i.e., during status epilepticus, gradually reduced electrical and behavioral seizure activity and greatly enhanced the survival rate. These results suggest that activation of NMDA receptors plays an important role in status epilepticus and brain damage in the lithium-pilocarpine model. This was further supported by results showing that nonconvulsive doses of NMDA and pilocarpine were synergistic, resulting in status epilepticus and subsequent mortality.

Sodium nitroprusside and guanosine 3',5'-monophosphate (cyclic GMP) inhibit stimulated phosphoinositide hydrolysis in rat cerebral cortical slices.

The combination of carbachol and 20 mM K+ greatly stimulates phosphoinositide hydrolysis in rat cerebral cortical slices. This response was inhibited by sodium nitroprusside, an activator of guanylate cyclase which elevated guanosine 3',5'-monophosphate (cyclic GMP) levels. 8-Bromocyclic GMP and the phorbol ester phorbol myristate acetate (PMA) also inhibited phosphoinositide hydrolysis, and the inhibitory effect of PMA was additive with that of sodium nitroprusside. As guanylate cyclase is Ca2+-dependent this inhibition of phosphoinositide hydrolysis by cyclic GMP may serve as a feedback modulator of the production of inositol phosphates and the ensuing rise of intracellular calcium.