Rescuing prefrontal cAMP-CREB pathway reverses working memory deficits during withdrawal from prolonged alcohol exposure.

Both human and animal studies indicate that alcohol withdrawal following chronic alcohol consumption (CAC) impairs many of the cognitive functions which rely on the prefrontal cortex (PFC). A candidate signaling cascade contributing to memory deficits during alcohol withdrawal is the protein kinase A (PKA)/cAMP-responsive element binding (CREB) cascade, although the role of PKA/CREB cascade in behavioral and molecular changes during sustained withdrawal period remains largely unknown. We demonstrated that 1 week (1W) or 6 weeks (6W) withdrawal after 6-month CAC impairs working memory (WM) in a T-maze spontaneous alternation task and reduces phosphorylated CREB (pCREB) in the PFC but not the dorsal CA1 region (dCA1) of the hippocampus compared with CAC and water conditions. In contrast, both CAC-unimpaired and withdrawn-impaired mice exhibited decreased pCREB in dCA1 as well as reduced histone H4 acetylation in PFC and dCA1, compared with water controls. Next, we showed that enhancing CREB activity through rolipram administration prior to testing improved WM performance in withdrawn mice but impaired WM function in water mice. In addition, WM improvement correlates positively with increased pCREB level selectively in the PFC of withdrawn mice. Results further indicate that direct infusion of the PKA activator (Sp-cAMPS) into the PFC significantly improves or impairs, respectively, WM performance in withdrawn and water animals. In contrast, Sp-cAMPS had no effect on WM when infused into the dCA1. Collectively, these results provide strong support that dysregulation of PKA/CREB-dependent processes in prefrontal neurons is a critical molecular signature underlying cognitive decline during alcohol withdrawal.

[Screening of the risk of functional decline performed by an inpatient geriatric consultation team in a general hospital]. / Dépistage du risque de déclin fonctionnel par une equipe gériatrique mobile au sein d'un hôpital général.

The Mobile Geriatric Team (MGT) is part of the Geriatric Care Program and aims to provide interdisciplinary geriatric expertise to other professionals for old patients hospitalized outside geriatric department. Our hospital has a MGT since 2008. Our objective is to retrospectively describe the population of patients of 75 years and older hospitalized outside the geriatric ward and screened for the risk of functional decline by the MGT between 1 October 2009 and 30 September 2011. We recorded the risk of functional decline, as indicated by the Identification of Senior At Risk score (ISAR) performed within 48 h after admission, place of living, discharge destination, Mini Mental State Examination (MMSE) and Geriatric Depression Scale (GDS) scores. In two years, 1.568 patients > or = 75 Y were screened with the ISAR score (mean age 82.5 Y, 60.7% of women). We identified 833 patients with a high-risk of functional decline (ISAR > or = 3). The majority of high-risk subjects (78%) were living at home before hospitalization and 58.7% returned home after discharge. Depression and cognitive impairment were identified among respectively 41% and 59% of high-risk subjects. Only 128 patients were admitted for fall. Most of the faller patients were living at home prior hospitalization and had an ISAR score > or = 3. The MGT allowed identifying many patients > or = 75 Y living at home and presenting with high-risk of functional decline and geriatric syndromes, confirming that good screening procedures are necessary to optimize management of hospitalized olders. Most of faller patients have an ISAR score > or = 3 and should benefit a comprehensive geriatric assessment.

Spatial learning induces differential changes in calcium/calmodulin-stimulated (ACI) and calcium-insensitive (ACII) adenylyl cyclases in the mouse hippocampus.

Several lines of evidence indicate that Ca2+/calmodulin-stimulated isoforms of adenylyl cyclase (AC) are involved in long-term potentiation and in certain forms of learning. Recently, we found that training in different types of learning task differentially activates Ca2+-sensitive versus Ca2+-insensitive AC activities in certain brain regions, indicating that AC species other than those stimulated by Ca2+/calmodulin may play an important role in learning processes (Guillou, Rose, & Cooper, 1999). Here, we report the effects of spatial reference memory training in a radial arm maze on the levels of AC1 and AC2 mRNA in the dorsal hippocampus of C57BL/6 mice. Acquisition of the task was associated with a learning-specific and time-dependent increase of AC1 mRNA expression selectively in subfields CA1-CA2. In contrast, AC2 mRNA levels were either reduced or not reliably affected depending on the stage of acquisition. Moreover, no significant changes in AC expression were observed either in the dorsal hippocampus of mice trained in a non-spatial (procedural) version of the task or in cortical regions of mice learning the spatial or procedural task. The regional specificity of these effects indicates that the formation of spatial and non-spatial memory requires distinct contributions from Ca2+-sensitive and Ca2+-insensitive AC in the hippocampus. It is suggested that downregulation of AC2 throughout all hippocampal subfields may play a permissive role during the acquisition of spatial learning whereas an upregulation of AC1 specifically in subfield CA1, may be critical to accurately encode, store or use spatial information.

Deficits of spatial and non-spatial memory and of auditory fear conditioning following anterior thalamic lesions in mice: comparison with chronic alcohol consumption.

This study was aimed at determining (i) whether or not bilateral subtotal lesions of the anterior thalamic nuclei (ATH) in rodents produced memory deficits for spatial and/or non-spatial information and of auditory fear conditioning, and (ii) if these eventual deficits resemble those produced by chronic alcohol consumption (CAC). Working memory was assessed using both spatial (spontaneous alternation) and non-spatial (temporal alternation) delayed response tasks. Results showed that ATH lesions induced delay-dependent memory impairments in both spatial and non-spatial alternation tasks, as well as a decreased level of auditory and background contextual fear conditioning compared with respective controls. CAC did not induce accelerated rate of forgetting in the spatial and non-spatial tasks, but increased the vulnerability to interference in the spatial task. CAC impaired only background contextual fear conditioning. We conclude that ATH nuclei are involved in the maintenance of information over time, regardless of the nature (spatial vs. non-spatial) of the information, and play a role in associative processes for both unimodal (the tone) and polymodal (contextual) information. In contrast, ATH dysfunction does not account for the memory disorders induced by the CAC treatment. Our results contribute to showing that the functional overlap between the structures comprising the hippocampo-mamillo-thalamic pathway is only partial.

Differential regulation of Ca(2+)-calmodulin stimulated and Ca(2+)-insensitive adenylyl cyclase messenger RNA in intact and denervated mouse hippocampus.

The Ca(2+)-calmodulin stimulated AC1 and Ca(2+)-insensitive AC2 are major isoforms of adenylyl cyclase, playing an important role in synaptic plasticity in the mammalian brain. We studied the pattern of expression of AC1 and AC2 genes in the hippocampus of C57BL/6 mice. We found that there were differences in their patterns of distribution in the dentate gyrus. AC1 messenger RNA was detected both in the dentate granule cell bodies and the corresponding molecular field whereas AC2 messenger RNA was preferentially distributed in the dentate granule cell layer, suggesting that AC1 and AC2 messenger RNA are differentially regulated in the dentate gyrus. In order to examine the regulation of AC1 and AC2 expression in response to synaptic deafferentation and reinnervation, the distribution patterns of the two AC messenger RNA in the hippocampal fields and the parietal cortex were analysed 2, 5, 9 and 30 days following an unilateral entorhinal cortex lesion. Interestingly, we found significantly reduced levels of AC1 hybridization signal following the lesion whereas the level of AC2 messenger RNA remained unaffected in all lesioned groups. The changes in AC1 messenger RNA were transient, with a maximal reduction at five days postlesion, and were restricted to the granule cell bodies and stratum moleculare of the deafferented dentate gyrus. No significant change in AC1 messenger RNA levels was detected in other hippocampal fields nor for any other postlesion times studied. These findings suggest that, at least in the dentate gyrus, messenger RNA for AC1 and AC2 might be differentially compartmentalized in cell bodies and dendritic fields. The activity-dependent regulation of AC1 messenger RNA levels by afferent synapses may provide an elegant mechanism for achieving a selective local regulation of AC1 protein, close to its site of action.

Ca2+-sensitive adenylyl cyclases, key integrators of cellular signalling.

The concept of second messenger signalling originated from the discovery of the role of cyclic AMP, although it is now known that cytosolic calcium [Ca2+]i mediates numerous signalling pathways and plays an equally vital role in many cellular events. In the last few years there has been a great deal of interest in the substantial molecular and functional diversity of mammalian adenylyl cyclases (ACs). Although AC was viewed as a generic activity, which was either stimulated or inhibited by stimulatory or inhibitory receptors, respectively, acting via alpha-subunits of trimeric GTP-regulatory proteins, the recent cloning of nine full-length isoforms, which significantly differ in their regulatory properties and tissue distributions, has revealed an unexpected level of complex regulation. In fact, each AC may integrate convergent inputs from many distinct signal-generating pathways. The nine isoforms can be divided into four distinct families, which reflect their distinct patterns of regulation by betagamma subunits of G-proteins, protein kinase C (PKC) and Ca2+. The mechanisms of regulation are often highly synergistic or conditional, suggesting a function of ACs as coincident detectors. Since all nine isoforms can be regulated either directly or indirectly by Ca2+ or PKC, a complex range of responses is possible. The Ca2+ concentration that stimulates the major ACs in brain has been found to inhibit AC activity in a number of peripheral tissues and cell lines. The purpose of this article is to review many of the important aspects about the distinct regulatory properties and cellular distribution of Ca2+-regulated ACs. Indeed, the notion that Ca2+ and cAMP are "synarchic" messengers acting in concert to regulate cellular activity was formally proposed some time ago. Here, we will focus on acute interactions between Ca2+ and cAMP and attempt to understand how AC activities can be regulated by discrete, physiological [Ca2+]i rises in intact cells. All Ca2+-regulated isoforms have characteristic distribution patterns in the brain. Also discussed are emerging insights on the temporal and spatial regulation of Ca2+- and cAMP-regulated pathways which may enable cell stimuli to elicit specific responses.

Down syndrome-critical region contains a gene homologous to Drosophila sim expressed during rat and human central nervous system development.

Many features of Down syndrome might result from the overdosage of only a few genes located in a critical region of chromosome 21. To search for these genes, cosmids mapping in this region were isolated and used for trapping exons. One of the trapped exons obtained has a sequence very similar to part of the Drosophila single-minded (sim) gene, a master regulator of the early development of the fly central nervous system midline. Mapping data indicated that this exonic sequence is only present in the Down syndrome-critical region in the human genome. Hybridization of this exonic sequence with human fetal kidney poly(A)+ RNA revealed two transcripts of 6 and 4.3 kb. In situ hybridization of a probe derived from this exon with human and rat fetuses showed that the corresponding gene is expressed during early fetal life in the central nervous system and in other tissues, including the facial, skull, palate, and vertebra primordia. The expression pattern of this gene suggests that it might be involved in the pathogenesis of some of the morphological features and brain anomalies observed in Down syndrome.

Individual differences in multiple-bag spindles of cat superficial lumbrical muscles.

A total of 791 spindle poles was analysed with regard to intrafusal fibre composition in the first and second superficial lumbrical muscles from the right and left hindfeet of 9 male and 5 female adult cats. Bag and chain muscle fibres were identified by their myofibrillar ATPase staining profile in the B region, after either acid or alkaline preincubation. A high proportion of the spindle pole population (43.2%) was observed to contain three or more (up to 5) bag fibres; those poles were classified as multiple-bag spindle poles. In the 334 muscle spindles in which both poles were studied, 42 bag fibres (12.6%) were found to be of the 'mixed' type, that is a fibre in which the two poles differ in their ATPase staining profile (either bag1/bag2 or bag/chain). The variability of the intrafusal fibre content observed in spindles of these muscles has been studied in relation to individual characteristics such as sex, weight and side of the animal. In general, multiple-bag spindles are more frequent in male than in female cats and in right as compared to left side muscles. Nearly all 'mixed' bag intrafusal fibres (38 out of 42) were observed in spindles containing 3 or more bag fibres. In 3-bag spindles the proportion of 'mixed' bag spindles is approximately the same in male and female cats. The ratio of 'dynamic' (mean polar bag1 content) to 'static' (mean polar bag2 plus chain fibre content) intrafusal effectors per muscle tends to increase in spindles of right side muscles and to decrease in the heaviest animals. The quantitative and qualitative differences in fibre content of spindles observed in first lumbrical muscles of different animals suggest that the spindle fibre composition, especially that of the 'dynamic' bag1 fibre, may be related to individual predetermined and/or acquired factors.

High incidence of long-chain fibers in spindles of cat superficial lumbrical muscles.

Forty-two complete spindle poles of cat superficial lumbrical muscles were analyzed with particular regard to the length and the diameter of intrafusal fiber types. Poles were reconstructed from serial transverse sections of fresh-frozen muscles. The staining module, which was repeated throughout the whole muscle, comprised sections treated for glycogen detection and sections treated for detecting myofibrillar ATPase activity after preincubation at three different pH's (see METHODS). The identification of intrafusal fiber types was essentially based on the ATPase activity of the B region of the intrafusal fibers. Long-chain fibers, i.e., chain fibers that have at least one pole that extends by more than one millimeter beyond the end of the spindle capsule (6), were very commonly observed. Of 42 spindle poles analyzed, 30 (71%) contained at least one long-chain fiber (one in 17 spindle poles, 2 in 11 poles, and 3 in 2 poles). Of 246 poles of chain fibers, 45 (18%) were "long". In four spindles, in which both poles could be completely examined, 10 long-chain fibers were observed. In eight of these, only one pole was long; the opposite pole ended either intracapsularly or at a short distance outside the capsule. Since long-chain fiber poles, presently considered to be among the effectors of static skeletofusimotor (beta) axons, are present in a large proportion of muscle spindles of lumbrical muscles, it would be of particular interest to reevaluate the beta-supply of these muscles by physiological methods.

[Glycogen depletion produced in intrafusal nuclear bag muscle fibers by brief large-amplitude muscle stretches]. / Déplétion glycogénique produite dans les fibres musculaires intrafusales à sac nucléaire par des étirements musculaires brefs de forte amplitude.

The glycogen content of the three types of intrafusal muscle fibre was studied with histochemical techniques in cat muscle spindles of superficial lumbrical muscles after a very large number of brief large stretches. Zones of glycogen depletion were observed in a high proportion of nuclear bag fibres, notably in bag 1 fibres, but not in chain fibres. These observations suggest that stretching of bag fibres by itself may activate these fibres.

Glycogen depletion elicited in tenuissimus intrafusal muscle fibres by stimulation of static gamma-axons in the cat.

In this study the experimental conditions used to elicit glycogen depletion in tenuissimus intrafusal muscle fibres were different from those used by Barker, Emonet-Dénand, Harker, Jami & Laporte (1976): the tenuissimus was left in situ; several (4-6) static gamma-axons were stimulated together; the blood flow through the muscle was not reduced during the periods of gamma stimulation except in two experiments; very much longer periods (up to 9 h) of intermittent stimulation by bursts at 50-500/s were used. Bag1 and bag2 fibres were identified by their different ATPase activities in the B region. In two experiments with normal circulation, test responses of several primary endings to short periods of stimulation at 50-100/s were still very strong after stimulation of several static gamma-axons for 5 and 9 h, respectively. Glycogen depletion was observed in a large number of chain and bag2 poles but in only one of nineteen bag1 poles examined. In two other experiments with normal circulation, there was a very pronounced reduction of the test responses after stimulation of several static gamma-axons for 7 and 9 h, respectively. Out of twenty-four bag1 poles examined, nineteen exhibited zones of depletion. In an experiment in which stimulation was conducted as in Barker et al. (1976), i.e. with reduction of muscle blood flow during 1 min periods of stimulation at 50-100/s, the primary endings still gave a strong response after fifteen periods of stimulation in contrast with the marked 'fatigue' that was constantly observed in the former study. No depleted intrafusal fibres were found in the spindles of this muscle. In a last experiment, after an initial pattern of stimulation similar to that described above, the new pattern of stimulation, but with periodical reduction of blood flow, was applied, leading to a 'fatigue' of the test responses in 2 h. In the spindles of this muscle three out of ten bag1 poles were depleted. The variability of glycogen depletion in bag1 fibres appears to be linked to the degree of spindle 'fatigue' which may develop after static gamma stimulation. It seems that in 'fatigued' spindles some factor or factors liberated by the contraction of neighbouring fibres may deplete glycogen in bag1 fibres by a non-neural mechanism. When, in spite of a prolonged stimulation of static gamma-axons, no fatigue of the test responses develops, zones of depletion in bag1 fibres--possibly of neural origin--are very rare, although a large proportion of bag2 and chain fibres are depleted.