Hippocampal Cholinergic Mechanisms Research Articles

Neurokinin1 − cholinergic receptor mechanisms in the medial Septum-Dorsal hippocampus axis mediates experimental neuropathic pain

The neurokinin-1 receptors (NK1Rs) in the forebrain medial septum (MS) region are localized exclusively on cholinergic neurons that partly project to the hippocampus and the cingulate cortex (Cg), regions implicated in nociception. In the present study, we explored the hypothesis that neurotransmission at septal NK1R and hippocampal cholinergic mechanisms mediate experimental neuropathic pain in the rodent chronic constriction injury model (CCI). Our investigations showed that intraseptal microinjection of substance P (SP) in rat evoked a peripheral hypersensitivity (PH)-like response in uninjured animals that was attenuated by systemic atropine sulphate, a muscarinic-cholinergic receptor antagonist. Conversely, pre-emptive destruction of septal cholinergic neurons attenuated the development of pH in the CCI model that also prevented the expression of cellular markers of nociception in the spinal cord and the forebrain. Likewise, anti-nociception was evoked on intraseptal microinjection of L-733,060, an antagonist at NK1Rs, and on bilateral or unilateral microinjection of the cholinergic receptor antagonists, atropine or mecamylamine, into the different regions of the dorsal hippocampus (dH) or on bilateral microinjection into the Cg. Interestingly, the effect of L-733,060 was accompanied with a widespread decreased in levels of CCI-induced nociceptive cellular markers in forebrain that was not secondary to behaviour, suggesting an active modulation of nociceptive processing by transmission at NK1R in the medial septum. The preceding suggest that the development and maintenance of neuropathic nociception is facilitated by septal NK1R-dH cholinergic mechanisms which co-ordinately affect nociceptive processing in the dH and the Cg. Additionally, the data points to a potential strategy for pain modulation that combines anticholinergics and anti-NKRs.

Opposite Pathways of Cholinergic Mechanisms of Hypoxic Preconditioning in the Hippocampus: Participation of Nicotinic α7 Receptors and Their Association with the Baseline Level of Startle Prepulse Inhibition.

(1) Background. A one-time moderate hypobaric hypoxia (HBH) has a preconditioning effect whose neuronal mechanisms are not studied well. Previously, we found a stable correlation between the HBH efficiency and acoustic startle prepulse inhibition (PPI). This makes it possible to predict the individual efficiency of HBH in animals and to study its potential adaptive mechanisms. We revealed a bi-directional action of nicotinic α7 receptor agonist PNU-282987 and its solvent dimethyl sulfoxide on HBH efficiency with the level of PPI > or < 40%. (2) The aim of the present study was to estimate cholinergic mechanisms of HBH effects in different brain regions. (3) Methods: in rats pretested for PPI, we evaluated the activity of synaptic membrane-bound and water-soluble choline acetyltransferase (ChAT) in the sub-fractions of ‘light’ and ‘heavy’ synaptosomes of the neocortex, hippocampus and caudal brainstem in the intact brain and after HBH. We tested the dose-dependent influence of PNU-282987 on the HBH efficiency. (4) Results: PPI level and ChAT activity correlated negatively in all brain structures of the intact animals, so that the values of the latter were higher in rats with PPI < 40% compared to those with PPI > 40%. After HBH, this ChAT activity difference was leveled in the neocortex and caudal brainstem, while for membrane-bound ChAT in the ‘light’ synaptosomal fraction of hippocampus, it was reversed to the opposite. In addition, a pharmacological study revealed that PNU-282987 in all used doses and its solvent displayed corresponding opposite effects on HBH efficiency in rats with different levels of PPI. (5) Conclusion: We substantiate that in rats with low and high PPI two opposite hippocampal cholinergic mechanisms are involved in hypoxic preconditioning, and both are implemented by forebrain projections via nicotinic α7 receptors. Possible causes of association between general protective adaptation, HBH, PPI, forebrain cholinergic system and hippocampus are discussed.

Converging evidence that short-active photoperiod increases acetylcholine signaling in the hippocampus.

Seasonal variations in environmental light influence switches between moods in seasonal affective disorder (SAD) and bipolar disorder (BD), with depression arising during short active (SA) winter periods. Light-induced changes in behavior are also seen in healthy animals and are intensified in mice with reduced dopamine transporter expression. Specifically, decreasing the nocturnal active period (SA) of mice increases punishment perseveration and forced swim test (FST) immobility. Elevating acetylcholine with the acetylcholinesterase inhibitor physostigmine induces depression symptoms in people and increases FST immobility in mice. We used SA photoperiods and physostigmine to elevate acetylcholine prior to testing in a probabilistic learning task and the FST, including reversing subsequent deficits with nicotinic and scopolamine antagonists and targeted hippocampal adeno-associated viral administration. We confirmed that physostigmine also increases punishment sensitivity in a probabilistic learning paradigm. In addition, muscarinic and nicotinic receptor blockade attenuated both physostigmine-induced and SA-induced phenotypes. Finally, viral-mediated hippocampal expression of human AChE used to lower ACh levels blocked SA-induced elevation of FST immobility. These results indicate that increased hippocampal acetylcholine neurotransmission is necessary for the expression of SA exposure-induced behaviors. Furthermore, these studies support the potential for cholinergic treatments in depression. Taken together, these results provide evidence for hippocampal cholinergic mechanisms in contributing to seasonally depressed affective states induced by short day lengths.

Deficits in Water Escape Performance and Alterations in Hippocampal Cholinergic Mechanisms Associated With Neonatal Monosodium Glutamate Treatment in Mice

Mice treated neonatally with monosodium glutamate (MSG) were found to have learning and memory deficits in performing a non-spatial water escape task. Scopolamine impaired the water-escape performance of the control mice but not that of the MSG-treated mice. It was suggested that the water-escape performance deficit in the MSG-treated mice was a result of impaired central cholinergic mechanisms. As such, scopolamine was unable to further incapacitate an already impaired cholinergic system. This is strongly suppported by the decreased affinity of the sodium-dependent high-affinity choline uptake observed in the hippocampus. d-Cycloserine, a partial agonist at the glycine site of the NMDA receptor, did not affect the water-escape performance of the MSG-treated and control mice; nor did it alter the effects of scopolamine. This lack of effect of d-Cycloserine may imply that the NMDA receptors are not involved in non-spatial learning, in contrast to their reported involvement in spatial learning.

Enhancement of acetylcholine release by flumazenil in the hippocampus of rats chronically treated with diazepam but not with imidazenil or abecarnil.

The effects of long-term treatment (three times a day for 3 weeks) with pharmacologically active doses of the novel anxiolytics and anticovulsants abecarnil (0.5 mg/kg, IP) and imidazenil (0.5 mg/kg, IP) on basal hippocampal acetylcholine release in freely moving rats were compared with those of diazepam (3 mg/kg, IP). Challenge doses of diazepam, abecarnil, and imidazenil decreased the extracellular acetyl-choline concentration in the hippocampus by the same extent in animals chronically treated with the respective drug or vehicle. Moreover, the abrupt discontinuation of long-term treatment with diazepam, abecarnil, or imidazenil failed to affect hippocampal acetylcholine release during the first 5 days of withdrawal. In contrast, the acute administration of the benzodiazepine receptor antagonist flumazenil (1 mg/kg, IP) 2 days after diazepam withdrawal elicited a marked increase (65%) in acetylcholine release in the hippocampus. Flumazenil failed to induce the same effect 5 days after diazepam withdrawal or 2 or 5 days after discontinuation of long-term treatment with abecarnil or imidazenil. These results indicate that (i) the inhibitory effects of full (diazepam), partial (imidazenil), and selective (abecarnil) benzodiazepine receptor agonists on acetylcholine output in rat hippocampus are not affected by repeated drug administration; (ii) discontinuation of long-term treatment with each type of agonist does not affect hippocampal cholinergic mechanisms; and (iii) flumazenil increases acetylcholine release only in the hippocampus of rats chronically treated with diazepam. Together, these data further differentiate the pharmacology of benzodiazepine receptor full agonists from that of partial and selective agonists.

Increases in hippocampal and frontal cortical acetylcholine release associated with presentation of sensory stimuli

In vivo microdialysis was employed to monitor acetylcholine release in the hippocampus and frontal cortex of freely behaving rats. Four stimuli were presented on separate occasions in the course of a dialysis session to rats with microdialysis probes implanted in the hippocampus or frontal cortex. Visual, auditory, olfactory and tactile stimuli elicited a number of different responses such as exploratory and consummatory behaviours. Presentation of two of the stimuli (auditory and tactile) also produced periods of alert immobility (freezing). All of the stimuli increased acetylcholine release in both the hippocampus and cortex: in the hippocampus, this increase was statistically significant with all except the olfactory stimulus, whereas in the cortex all but the visual stimulus resulted in significant increases. In the hippocampus, there were no significant differences between the increases in acetylcholine release produced by the four stimuli. In contrast, in the cortex, there was significant variation between the magnitude of acetylcholine release produced by the different stimuli: acetylcholine release elicited by tactile stimulation was greater than that produced by the other stimuli. There was no significant variation in the duration of increases in acetylcholine release produced by the stimuli in either the hippocampus or cortex. These results provide evidence that acetylcholine release is associated with a variety of behavioural responses to stimuli designed to produce arousal, and point to a role for cortical and hippocampal cholinergic mechanisms in arousal or attention. Further, the results suggest that under some circumstances cortical and hippocampal acetylcholine release may be regulated differentially.

Effect of pyroglutamic acid stereoisomers on ecs and scopolamine-induced memory disruption and brain acetylcholine levels in the rat

The acquisition of a passive avoidance conditioned response was disrupted in the rat by electroconvulsive shock (ECS) and scopolamine administration. D,L-pyroglutamic acid (D,L-PCA) 500 and 1000 mg/kg, administered as arginine salt 120 min before the retest, prevented both the ECS and scopolamine-induced amnesia. Arginine alone was ineffective. Scopolamine brought about a 52 and 39% decrease, respectively, in cortical and hippocampal acetylcholine (ACh) levels, measured by means of a gas-chromatographic method. D,L-PCA 500 and 1000 mg/kg also prevented the decrease in brain ACh level. When the two isomers were studied separately, D-PCA was more effective than L-PCA and antagonized scopolamine-induced amnesia at the doses of 250 and 500 mg/kg. In conclusion, D,L-PCA is active on cortical and hippocampal cholinergic mechanisms and, like other 2-oxopy colidone derivatives, shows cognition-enhancing properties.

Hippocampal muscarinic cholinergic mediation of spontaneous alternation and fear in the developing rat.

Spontaneous alternation was examined in young rats following microinjections of anticholinergic agents into 4 hippocampal sites: anterodorsal or posteroventral dentate gyrus, hippocampus or entorhinal complex. The rate of alternation remained around 40% at 5, 10, and 15 days, increased suddenly to 80% on day 17, did not vary until day 20, regressed partly and temporarily until day 30, and returned to a near-adult level on day 40. Concomitantly with the transient regression of alternation between days 20 and 40, fear responses to environment were seen to emerge (boluses and squeaks), to reach a maximum on day 30, and to return to a low level by day 40. Injections of atropine (4, 8 micrograms) or scopolamine (4, 10 micrograms) into any of the 4 sites significantly reduced the rate of alternation from day 17 on. Only the highest doses were active at 10 and 15 days. These results demonstrate that spontaneous alternation and hippocampal muscarinic cholinergic mechanisms develop simultaneously and progress suddenly on postnatal day 17. Atropine and scopolamine also affected fear responses, abolishing or potentiating them according to the site of injection, showing that hippocampal cholinergic mechanisms exert complex influences on fear-induced emotional reactions.

Oxiracetam prevents electroshock-induced decrease in brain acetylcholine and amnesia

In the rat, 1 min following electroshock (ECS) a 46 and 39% decrease in acetylcholine levels was found in the hippocampus and cerebral cortex, respectively. The decrease in the hippocampus was still statistically significant 30 min after ECS. The ECS applied 1 min after training also disrupted the performance of a passive avoidance conditioned response (‘step down’) tested 30 min later. Oxiracetam (100 and 300 mg/kg i.p.) administered 90 min before training prevented, in a dose-dependent manner, the decrease of acetylcholine in the cerebral cortex and hippocampus. Oxiracetam prevented the ECS disruption of the acquisition of a passive avoidance response. At the dose of 300 mg/kg the acetylcholine level 1 min after ECS was significantly higher than in the sham-treated rats. Piracetam at the same doses was inactive. These results support the hypothesis that oxiracetam may prevent the disruption of the conditioned response by acting on cortical and hippocampal cholinergic mechanisms.

Effects of hippocampal electrical stimulation on long-term memory and on cholinergic mechanisms in three inbred strains of mice

Two sets of experiments have been carried out in an attempt to determine the role of hippocampal cholinergic mechanisms in a long-term memory storage. Three inbred strains of mice were presented with two different learning tasks in order to estimate their long-term retention abilities as well as changes in this ability after a post-trial hippocampal stimulation. In parallel experiments the enzymes involved in acetylcholine metabolism were studied under different experimental conditions. Our results indicate: (a) The capacity for long-term memory of the BALB/c line is much greater than that of either the C57BL/6 or C57BR strain. (b) Hippocampal post-trial electrical stimulation leads to an improvement of this capacity in the BALB/c strain. This phenomenon is less pronounced in C57BL/6 and non-existent in C57BR mice. (c) Choline acetyltransferase activity in the hippocampus is significantly higher in BALB/c than in the other two strains. In BALB/c this enzyme activity is greatly changed by the post-trial stimulation whereas in the C57BL/L strain only a slight variation of enzyme activity is observed. No modification occurs in C57BR. The results suggest that the more active acetylcholine synthesizing enzyme in the hippocampus of BALB/c may be related to a greater acetylcholine availability, thus favoring the establishment of a long-term memory, perhaps by releasing greater amounts of acetylcholine in the hippocampus immmediately after the learning session. The electrical stimulation of the hippocampus acts to magnify or accelerate this phenomenon. It is suggested that the efficiency of the stimulation would be related to the genetically determined higher cholinergic activity of the hippocampus.

Effects of hippocampal and lateral hypothalamic electrical stimulation on hippocampal cholinergic mechanisms in BALB/c inbred mice

Electrical stimulation of the hippocampus (HPC) and of the lateral hypothalamus (LH) were found to have the same 24 h delayed effect on choline acetyltransferase activity. Since previous experiments demonstrated that these treatments had the same effect on memory storage processes, the present finding and earlier neurochemical investigations suggest that HPC and LH stimulation might act through a common hippocampal cholinergic mechanism.

The role of hippocampal cholinergic mechanisms in the aquisition of a barpress response

The role of hippocampal cholinergic mechanisms in learning a bar-press response reinforced with food was investigated. Firstly, an interstrain comparison showed that mice having a low choline acetyltransferase activity in the dorsal hippocampus were quicker to associate the barpress with reiforcement. Secondly, when the activity of this enzyme was reduced by a subseizure electrical stimulation of the hippocampus learning was accelerated. It is suggested that acetylcholine availability at the hippocampal synapses showed the apparition of these learned responses.