Ageing is accompanied by a decline in cognitive functions; along with a variety of neurobiological changes. The association between inflammation and ageing is based on complex molecular and cellular changes that we are only just beginning to understand. The hippocampus is one of the structures more closely related to electrophysiological, structural and morphological changes during ageing. In the present study we examined the effect of normal ageing and LPS-induced inflammation on astroglia-neuron interaction in the rat hippocampus of adult, normal aged and LPS-treated adult rats. Astrocytes were smaller, with thicker and shorter branches and less numerous in CA1 Str. radiatum of aged rats in comparison to adult and LPS-treated rats. Astrocyte branches infiltrated apoptotic neurons of aged and LPS-treated rats. Cellular debris, which were more numerous in CA1 of aged and LPS-treated rats, could be found apposed to astrocytes processes and were phagocytated by reactive microglia. Reactive microglia were present in the CA1 Str. Radiatum, often in association with apoptotic cells. Significant differences were found in the fraction of reactive microglia which was 40% of total in adult, 33% in aged and 50% in LPS-treated rats. Fractalkine (CX3CL1) increased significantly in hippocampus homogenates of aged and LPS-treated rats. The number of CA1 neurons decreased in aged rats. In the hippocampus of aged and LPS-treated rats astrocytes and microglia may help clearing apoptotic cellular debris possibly through CX3CL1 signalling. Our results indicate that astrocytes and microglia in the hippocampus of aged and LPS-infused rats possibly participate in the clearance of cellular debris associated with programmed cell death. The actions of astrocytes may represent either protective mechanisms to control inflammatory processes and the spread of further cellular damage to neighboring tissue, or they may contribute to neuronal damage in pathological conditions.

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1 The effects of histamine and agents acting at histamine receptors on spontaneous and 100 mM K+-evoked release of acetylcholine, measured by microdialysis from the cortex of freely moving rats, and on cognitive tests are described. 2 Local administration of histamine (0.1-100 pM) failed to affect spontaneous but inhibited 100 mM K+-stimulated release of acetylcholine up to about 50%. The H3 receptor agonists (R)-ca-methylhistamine (RAMH) (0.1-10 yM), imetit (0.01-10 yM) and immepip (0.01-10 yM) mimicked the effect of histamine.3 Neither 2-thiazolylethylamine (TEA), an agonist showing some selectivity for H, receptors, nor the H2 receptor agonist, dimaprit, modified 100 mM K+-evoked release of acetylcholine. 4 The inhibitory effect of 100 pM histamine was completely prevented by the highly selective histamine H3 receptor antagonist, clobenpropit but was resistant to antagonism by triprolidine and cimetidine, antagonists at histamine H, and H2 but not H3 receptors.5 The H3 receptor-induced inhibition of K+-evoked release of acetylcholine was fully sensitive to tetrodotoxin (TTX).6 The effects of intraperitoneal (i.p.) injection of imetit (5 mg kg-') and RAMH (5 mg kg-') were tested on acetylcholine release and short term memory paradigms. Both drugs reduced 100 mM K+-evoked release of cortical acetylcholine, and impaired object recognition and a passive avoidance response.7 These observations provide the first evidence of a regulatory role of histamine H3 receptors on cortical acetylcholine release in vivo. Moreover, they suggest a role for histamine in learning and memory and may have implications for the treatment of degenerative disorders associated with impaired cholinergic function.

Peptides present in the skin secretion of the South African frog, Xenopus laevis, have been analysed by fast atom bombardment mass spectrometry and h.p.l.c. in the mass range 500-3200 Da. We have investigated the effects of successive glandular secretions induced by noradrenaline injections on these peptide levels and have found that the replenishment of the whole range of peptides is complete within 2-6 days. Intact secretory vesicles free of cellular contaminants contain a relatively large number of peptides with molecular masses in the range 2400-2700 Da. We have termed these peptides primary products or spacer peptides, since they originate from spacer regions of the precursors to xenopsin and caerulein. However, if the secretory vesicles are disrupted during the collection procedure and the solution containing the secretion is kept at room temperature for up to 2 h, relatively little of the larger peptides remain. By comparing the relative levels of the various peptides present in these secretions we have found that the larger peptides are proteolytically cleaved into smaller fragments by a novel cleavage at the N-terminal side of a lysine residue (at Xaa-Lys bonds where Xaa is Leu, Gly, Ala or Lys). Preliminary evidence has been obtained suggesting that the larger intact peptides possess lytic activity whereas the smaller proteolytic fragments appear relatively inactive. This may represent a mechanism by which the secretions are rendered harmless to the frog itself, since prolonged exposure would be expected to result in toxic effects. The dorsal glands of X. laevis thus appear similar to endocrine glands, since they are involved in peptide biosynthesis, secretion and subsequent proteolytic degradation.

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Cholinesterase inhibitors (ChEIs) were introduced in the therapy of Alzheimer Disease (AD) in the nineteen nineties with great expectations. The hopes and large interest raised by these drugs are well demonstrated by 12,000 references listed by PubMed under 'ChEI' for 1995-2007. The list is reduced to 2500 if we confine ourselves to 'ChEIs and dementia'. Of them, about 500 were published in the last two years. Whereas an increase in brain acetylcholine and an improvement of cognitive deficits have been consistently demonstrated in animal models of AD, from aging rats to transgenic mice, the clinical effectiveness of ChEIs has been and is still a matter of contrasting opinions. These range from the negative conclusions of the AD2000 trial on donepezil, claiming that it is not cost effective, with benefits below a minimally relevant threshold, to the NICE appraisal of 2007 declaring that donepezil, rivastigmine, galantamine are efficacious for mild to moderate AD, irrespective of their different selectivity for acetyl- (AChE) and butyrylcholinesterase (BuChE). The possibility that ChEIs may exert their effects through mechanisms beyond cholinesterase inhibition has been envisaged. However, according to the information presented in this review, the "classical" ChEIs, donepezil, rivastigmine and galantamine, show no pharmacological actions beyond cholinesterase inhibition which may play an important role in their therapeutic efficacy. The diverging opinions on clinical efficacy do not discourage from developing new ChEIs, and particularly the so called multifunctional ChEIs. They represent the future of the cholinergic therapy for AD but other indications for these drugs may be considered, including vascular dementia, mild cognitive impairment, and the ethically sensitive improvement of memory and learning in healthy subjects.

Measuring the changes in neurotransmitter extracellular levels in discrete brain areas is considered a tool for identifying the neuronal systems involved in specific behavioral responses or cognitive processes. Acetylcholine (ACh) is the first neurotransmitter whose diffusion from the central nervous system was investigated and whose extracellular levels variations were correlated to changes in neuronal activity. This was done initially by means of the cup technique and then by the microdialysis technique. The latter, notwithstanding some technical limitations, makes it possible to detect variations in extracellular levels of ACh in unrestrained, behaving animals. This review summarizes and discusses the results obtained investigating the changes in ACh release during performance of operant tasks, exposition to novel stimuli, locomotor activity, and the performance of spatial memory tasks, working memory, and place preference memory tasks. Activation of the forebrain cholinergic system has been demonstrated in many tasks and conditions in which the environment requires the animal to analyze novel stimuli that may represent a threat or offer a reward. The sustained cholinergic activation, demonstrated by high levels of extracellular ACh observed during the behavioral paradigms, indicates that many behaviors occur within or require the facilitation provided by the cholinergic system to the operation of pertinent neuronal pathways.Acetylcholine (ACh) is the first neurotransmitter whose diffusion from the central nervous system was investigated and whose extracellular levels variations were correlated to changes in neuronal activity. ACh outflow from the spinal cord was detected by Bulbring and Burn (1941) during nerve stimulation, and the first attempts to demonstrate ACh outflow from the intact cortex were made 50 years ago. The purpose was to demonstrate whether ACh had a role in the CNS, and the theoretical approach was the same used in the previous decade for demonstrating the neurotransmitter role of ACh in the peripheral nervous system (Burn 1968). Using the cup technique and the leech bioassay, MacIntosh and Oboring (1955) demonstrated in the dog that ACh release was related to the spontaneous electrical activity of the cortex. The cup was formed by a small cylinder exerting a slight pressure over the meninges, filled with Ringer solution containing a cholinesterase inhibitor. No ACh could be detected in the absence of the inhibitor. The method was perfected by Mitchell (1953), and a description of the procedure with an analysis of the early literature reporting the changes in ACh release induced by stimulation of peripheral nerves, specific brain areas, and drugs can be found in reviews by Pepeu (1973), and Moroni and Pepeu (1984).The first question asked was whether ACh diffusing from the brain into the cortical cup originated from cholinergic nerve endings and whether changes in its extracellular levels were expressions of changes in the activity of the cholinergic nerve endings under the cup. The answer ...

All types of memory depend on the integrated activity of various brain structures and neurotransmitter systems and involve more than one receptor, signal transduction pathway and postsynaptic mechanism. The components of the extracellular signal regulated kinases-1 and -2 (ERK1/2) signal transduction pathways are ubiquitous and well conserved protein kinases involved in relaying extracellular signals into intracellular responses, and are involved in the mechanisms of synaptic plasticity, learning and memory. ERK activation is required for the full expression of long-term potentiation (LTP), the principal cellular mechanism thought to underlie neuronal plasticity. Furthermore, ERK is activated in and is necessary for the development of several forms of memory, such as fear conditioning, conditioned taste aversion memory, spatial memory, step-down inhibitory avoidance and object recognition memory. ERK activation is secondary to neurotransmitter release and activation of the forebrain cholinergic neurons during and immediately after acquisition of an inhibitory avoidance response, revealed by increased release of acetylcholine (ACh), which in turn activates ERK in neurons located in the medial prefrontal cortex and ventral hippocampus. Increased release of ACh and ERK activation are events mechanistically related to each other, as demonstrated by the use of scopolamine, a muscarinic receptor antagonist, and by inhibitors of ERK activation, which blocked memory encoding and ERK activation. A critical function of activated ERK downstream of the increased ACh release occurring during learning is to promote cellular integration of divergent downstream effectors which may trigger different responses, depending upon which subsets of scaffolding anchors, target proteins and regulatory phosphatases are involved. The hope is that by studying how ERK is activated by different neurotransmitter systems and the ensuing downstream cellular modifications, the molecular basis of memory will be ultimately understood.