Systemic oscillator-driven and nutrient-responsive hormonal regulation of daily expression rhythms for gluconeogenic enzyme genes in the mouse liver.

Gluconeogenesis is de novo glucose synthesis from substrates such as amino acids and is vital when glucose is lacking in the diurnal nutritional fluctuation. Accordingly, genes for hepatic gluconeogenic enzymes exhibit daily expression rhythms, whose detailed regulations under nutritional variations remain elusive. As a first step, we performed general systematic characterization of daily expression profiles of gluconeogenic enzyme genes for phosphoenolpyruvate carboxykinase (PEPCK), cytosolic form (Pck1), glucose-6-phosphatase (G6Pase), catalytic subunit (G6pc), and tyrosine aminotransferase (TAT) (Tat) in the mouse liver. On a standard diet fed ad libitum, mRNA levels of these genes showed robust daily rhythms with a peak or an elevation phase during the late sleep-fasting period in the diurnal feeding/fasting (wake/sleep) cycle. The rhythmicity was preserved in constant darkness, modulated with prolonged fasting, attenuated by Clock mutation, and entrained to varied photoperiods and time-restricted feedings. These results are concordant with the notion that gluconeogenic enzyme genes are under the control of the intrinsic circadian oscillator, which is entrained by the light/dark cycle, and which in turn entrains the feeding/fasting cycle and also drives systemic signaling pathways such as the hypothalamic-pituitary-adrenal axis. On the other hand, time-restricted feedings also showed that the ingestion schedule, when separated from the light/dark cycle, can serve as an independent entrainer to daily expression rhythms of gluconeogenic enzyme genes. Moreover, nutritional changes dramatically modified expression profiles of the genes. In addition to prolonged fasting, a high-fat diet and a high-carbohydrate (no-protein) diet caused modification of daily expression rhythms of the genes, with characteristic changes in profiles of glucoregulatory hormones such as corticosterone, glucagon, and insulin, as well as their modulators including ghrelin, leptin, resistin, glucose-dependent insulinotropic polypeptide (GIP), and glucagon-like peptide-1 (GLP-1). Remarkably, high-protein (60% casein or soy-protein) diets activated the gluconeogenic enzyme genes atypically during the wake-feeding period, with paradoxical up-regulation of glucagon, which frequently formed correlation networks with other humoral factors. Based on these results, we propose that daily expression rhythms of gluconeogenic enzyme genes are under the control of systemic oscillator-driven and nutrient-responsive hormones.

Effect of metal-containing topical agents on surface doses received during external irradiation.

The ability of topical metal-containing agents (MCAs) to enhance radiation dermatitis remains controversial. In the present study, we evaluated increases in surface doses associated with topical agents at different application thicknesses and with MCAs versus non-metal containing agents (NMCAs). We assessed two clinically available MCAs, zinc oxide ointment (ZOO) and silver sulfadiazine cream (SSDC), and eight NMCAs. Surface doses were measured using a Markus chamber placed on a polystyrene phantom. To evaluate the role of application thickness, each agent was applied to the chamber in oil-slick (<0.1-mm), 1-mm and 5-mm layers prior to irradiation of a 10 × 10 cm field with 4-, 6- and 10-MV X-ray beams. The surface dose enhancement ratio (SDER) was calculated as the ratio of the surface dose with an agent to the dose without an agent. The SDER values for the eight NMCAs, ZOO and SSDC at an oil-slick thickness were 101.6-104.6% (mean: 103.3%), 104.5% and 105.0%, respectively, using a 6-MV X-ray beam. The corresponding values at a 1-mm thickness were 196.8-237.8% (mean: 215.7%), 229.3% and 201.4%, respectively, and those at a 5-mm thickness were 342.2-382.4% (mean: 357.9%), 357.1% and 352.6%, respectively. A similar tendency was found using 4- and 10-MV X-ray beams. The lack of a significant difference in surface dose enhancement between MCAs and NMCAs, particularly when applied in oil-slick layers, suggests that MCAs do not need to be avoided or applied in a restricted manner during radiotherapy for dosimetric reasons.

Negative cues lead to more inefficient search than positive cues even at later stages of visual search.

Observers can focus their attention on task-relevant items in visual search when they have prior knowledge about the target's properties (i.e., positive cues). However, little is known about how negative cues, which specify the features of task-irrelevant items, can be used to guide attention away from distractors and how their effects differ from those of positive cues. It has been proposed that when a distractor color is cued, people would first select the to-be-ignored items early in search and then inhibit them later. The present study investigated how the effects of positive and negative cues differ throughout the visual search process. The results showed that positive cues sped up the early stage of visual search and that negative cues led to initial selection for inhibition. We further found that visual search with negative cues was more inefficient than that with positive cues even at later stages, suggesting that sustained inhibition is needed throughout the visual search process. Taken together, the results indicate that positive and negative cues have different functions: prior knowledge about target features can weight task-relevant information at early stages of visual search, and negative cues are used more inefficiently even at later stages of visual search.

Numerosity underestimation in sets with illusory contours.

People underestimate the numerosity of collections in which a few dots are connected in pairs by task-irrelevant lines. Such configural processing suggests that visual numerosity depends on the perceived scene segments, rather than on the perceived total area occupied by a collection. However, a methodology that uses irrelevant line connections may also introduce unnecessary distraction and variety, or obscure the perception of task-relevant items, given the saliency of the lines. To avoid such potentially confounding variables, we conducted four experiments where the line-connected dots were replaced with collinear inducers of Kanizsa-type illusory contours. Our participants had to compare two simultaneously presented collections and choose the more numerous one. Displays comprised c-shaped inducers and disks (Experiment 1), c-shaped inducers only (Experiments 2 and 4), or closed inducers (Experiment 3). One display always showed a 12- (Experiments 1-3) or 48-item reference pattern (Experiment 4); the other was a test pattern with numerosity varying between 9 and 15 (Experiments 1-3) or 36-60 items (Experiment 4). By manipulating the number of illusory contours in the test patterns, the level of connectedness increased or decreased respectively. The fitted psychometric functions revealed an underestimation that increased with the number of illusory contours in Experiments 1 and 2, but was absent in Experiments 3 and 4, where illusory contours were more difficult to perceive or larger numerosities were used. Results corroborate claims that visual numerosity estimation depends on segmented inputs, but only within moderate numerical ranges.

Electrophysiological evidence that top-down knowledge controls working memory processing for subsequent visual search.

Items in working memory guide visual attention toward a memory-matching object. Recent studies have shown that when searching for an object this attentional guidance can be modulated by knowing the probability that the target will match an item in working memory. Here, we recorded the P3 and contralateral delay activity to investigate how top-down knowledge controls the processing of working memory items. Participants performed memory task (recognition only) and memory-or-search task (recognition or visual search) in which they were asked to maintain two colored oriented bars in working memory. For visual search, we manipulated the probability that target had the same color as memorized items (0, 50, or 100%). Participants knew the probabilities before the task. Target detection in 100% match condition was faster than that in 50% match condition, indicating that participants used their knowledge of the probabilities. We found that the P3 amplitude in 100% condition was larger than in other conditions and that contralateral delay activity amplitude did not vary across conditions. These results suggest that more attention was allocated to the memory items when observers knew in advance that their color would likely match a target. This led to better search performance despite using qualitatively equal working memory representations.

Relative spatial frequency tuning and its contrast dependency in human perception.

Several physiological studies in cats and monkeys have reported that the spatial frequency (SF) tuning of visual neurons varies depending on the luminance contrast and size of stimulus. However, comparatively little is known about the effect of changing the stimulus contrast and size on SF tuning in human perception. In the present study, we investigated the effects of stimulus size and luminance contrast on human SF tuning using the subspace-reverse-correlation method. Measuring SF tunings at six different stimulus sizes and three different luminance contrast conditions (90%, 10%, and 1%), we found that human perception exhibits significant stimulus-size-dependent SF tunings. At 90% and 10% contrast, participants exhibited relative SF tuning (cycles/image) rather than absolute SF tuning (cycles/°) at response peak latency. On the other hand, at 1% contrast, the magnitude of the size-dependent-peak SF shift was too small for strictly relative SF tuning. These results show that human SF tuning is not fixed, but varies depending on the stimulus size and contrast. This dependency may contribute to size-invariant object recognition within an appropriate contrast rage.

[Effects of four bisphosphonates on macrophage phagocytosis: quantitative measurement by flow cytometry using high-fluorescence particles and human monocytic cell line THP-1].

Impairment of macrophage phagocytosis is a major cause of chronic inflammation. Bisphosphonates (BPs) are widely used as anti-osteoclastic agents. The effects of BPs on monocyte-macrophage lineage cells are being increasingly reported; however, the detailed effects of BPs on macrophage phagocytic activity are still unclear. We examined the effects of four BPs: clodronate as a non-nitrogen containing BP (non-N-BP), and pamidronate, alendronate, and zoledronate as nitrogen-containing BP(N-BP), on macrophage phagocytic activity. The uptake of high fluorescence-labeled polystyrene beads by the human monocytic cell line THP-1 was investigated by flow cytometry. All three N-BPs suppressed the phagocytosis of macrophages more potently than the non-N-BP, clodronate. Pamidronate and zoledronate were more potent than alendronate. BP induced the apoptosis of THP-1. Pamidronate and zoledronate induced apoptosis more effectively than clodronate. The method described to observe phagocytosis was simple and quantitative, and might be useful in screening for the effects of drugs, such as N-BP and non-N-BP, on phagocytic activity.

The secretogranin II gene is a signal integrator of glutamate and dopamine inputs.

Cooperative gene regulation by different neurotransmitters likely underlies the long-term forms of associative learning and memory, but this mechanism largely remains to be elucidated. Following cDNA microarray analysis for genes regulated by Ca(2+) or cAMP, we found that the secretogranin II gene (Scg2) was cooperatively activated by glutamate and dopamine in primary cultured mouse hippocampal neurons. The Ca(2+) chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester (BAPTA-AM) and the mitogen-activated protein kinase (MAPK) kinase (MEK) inhibitor PD98059 prevented Scg2 activation by glutamate or dopamine; thus, the Ca(2+) /MEK pathway is predicted to include a convergence point(s) of glutamatergic and dopaminergic signaling. Unexpectedly, the protein kinase A inhibitor KT5720 enhanced Scg2 activation by dopamine. The protein-synthesis inhibitor cycloheximide also enhanced Scg2 activation, and the proteasome inhibitor ZLLLH diminished the KT5720-mediated augmentation of Scg2 activation. These results are concordant with the notion that dopaminergic input leads to accumulation of a KT5720-sensitive transcriptional repressor, which is short-lived because of rapid degradation by proteasomes. This repression pathway may effectively limit the time window permissive to Scg2 activation by in-phase glutamate and dopamine inputs via the Ca(2+) /MEK pathway. We propose that the regulatory system of Scg2 expression is equipped with machinery that is refined for the signal integration of in-phase synaptic inputs. We proposed hypothetical mechanism for the regulation of the secretogranin II gene as a signal integrator of glutamate and dopamine inputs. Glutamate or dopamine activates the Ca(2+) /MEK/ERK pathway, which thus contributes to the signal integration. Concurrently, activation of the PKA inhibitor KT5720-sensitive pathway by dopamine leads to accumulation of the repressor protein X that is otherwise susceptible to proteasome degradation. This repression system may determine the time window permissive to the cooperative activation by in-phase glutamate and dopamine inputs.

Cueing effects of target location probability and repetition.

The search performance for targets is improved when the targets appear in a specific location more frequently than in other locations. Although this phenomenon, called the "probability cueing effect," has been reported in past studies, it is unclear whether probability cueing is driven by statistical learning and/or intertrial facilitation of the target location. We investigated the underlying mechanisms for probability cueing effects by manipulating probabilities and repetitions of the target appearance at each target location. The first experiment demonstrated that the reaction time benefits of both statistical learning and intertrial facilitation contributed to the probability cueing effect. In contrast, the second and third experiments demonstrated that the probability cueing effect did not occur when target location repetitions on consecutive trials were fully or partially restricted. Also, any intertrial facilitation effects disappeared if there were more than one intervening trials. These results suggest that consecutive target location repetitions throughout the experiment facilitate learning of the target location probability.

Time of day and nutrients in feeding govern daily expression rhythms of the gene for sterol regulatory element-binding protein (SREBP)-1 in the mouse liver.

Sterol regulatory element-binding protein-1 (SREBP-1) plays a central role in transcriptional regulation of genes for hepatic lipid synthesis that utilizes diet-derived nutrients such as carbohydrates and amino acids, and expression of SREBP-1 exhibits daily rhythms with a peak in the nocturnal feeding period under standard housing conditions of mice. Here, we report that the Srebp-1 expression rhythm shows time cue-independent and Clock mutation-sensitive circadian nature, and is synchronized with varied photoperiods apparently through entrainment of locomotor activity and food intake. Fasting caused diminution of Srebp-1 expression, while diabetic db/db and ob/ob mice showed constantly high expression with loss of rhythmicity. Time-restricted feedings during mid-light and mid-dark periods exhibited differential effects, the latter causing more severe damping of the oscillation. Therefore, "when to eat in a day (the light/dark cycle)," rather than "whenever to eat in a day," is a critical determinant to shape the daily rhythm of Srebp-1 expression. We further found that a high-carbohydrate diet and a high-protein diet, as well as a high-fat diet, cause phase shifts of the oscillation peak into the light period, underlining the importance of "what to eat." Daily rhythms of SREBP-1 protein levels and Akt phosphorylation levels also exhibited nutrient-responsive changes. Taken together, these findings provide a model for mechanisms by which time of day and nutrients in feeding shape daily rhythms of the Srebp-1 expression and possibly a number of other physiological functions with interindividual and interdaily differences in human beings and wild animals subjected to day-by-day changes in dietary timing and nutrients.

Phonological influences on lexicosemantic processing of kanji words.

To investigate the phonological influences on the lexicosemantic process with a strong orthographic constraint, we used kanji (morphogram) homophone words and measured, using magnetoencephalography, the neural activities during the silent reading of prime-target pairs. The primes were phonologically the same as or different from the targets or pseudocharacters. The neural activities in the left posterior temporal and inferior parietal areas became weaker with phonological repetition. Furthermore, stronger activities for the different condition in the left anterior temporal area and for the same condition in the left inferior frontal cortex, respectively, suggest the roles of these areas of the brain in the semantic processing of words and in the selection of appropriate meanings. We conclude that phonological information affects the lexicosemantic process even with a strong orthographic constraint.

Multifactorial regulation of daily rhythms in expression of the metabolically responsive gene spot14 in the mouse liver.

Spot14 is a putative transcriptional regulator for genes involved in fatty acid synthesis. The Spot14 gene is activated in response to lipogenic stimuli such as dietary carbohydrate and is also under circadian regulation. The authors investigated factors responsible for daily oscillation of Spot14 expression. If mice were kept under a 12-h light/12-h dark cycle with ad libitum feeding, Spot14 mRNA levels in the liver reached a peak at an early dark period when mice, as nocturnal animals, start feeding. Under fasting, while Spot14 mRNA levels were generally decreased, the rhythmicity was still maintained, suggesting contribution of both nutritional elements and circadian clock factors on robust rhythmicity of Spot14 expression. Effects of circadian clock factors were confirmed by the observations that the circadian rhythm of Spot14 expression was seen also under the constant darkness and that the rhythmicity was lost in Clock mutant mice. When mice were housed in short-photoperiod (6-h light/18-h dark) and long-photoperiod (18-h light/6-h dark) cycles, rhythms of Spot14 mRNA levels were phase advanced and phase delayed, respectively, being concordant with the notion that Spot14 expression is under the control of the light-entrainable oscillator. As for nutritional mediators, in the liver of db/db mice exhibiting hyperinsulinemia-accompanied hyperglycemia, Spot14 mRNA levels were constantly high without apparent rhythmicity, consistent with previous observations for strong activation of the Spot14 gene by glucose and insulin. Restricted feeding during the 4-h mid-light period caused a phase advance of the Spot14 expression rhythm. On the other hand, restricted feeding during the 4-h mid-dark period led to damping of the rhythmicity, apparently resulting from the separation of phases between effects of the light/dark cycle and feeding on Spot14 expression. Thus, the daily rhythm of Spot14 expression in the liver is under the control of the light-entrainable oscillator, food-entrainable oscillator, and food-derived nutrients, in a separate or cooperative manner.

Quantification of caveolin isoforms using quantitative real-time RT-PCR, and analysis of promoter CpG methylation of caveolin-1alpha in human T cell leukemia cell lines.

Caveolin-1, an essential structural component of caveolae, functions as a negative regulator for signal transduction and has been suggested to be a candidate tumor suppressor. Lack of caveolin-1 expression has been implicated in the pathogenesis of oncogenic cell transformation and tumorigenesis in many cancers. On the other hand, over-expression has also been associated with tumor progression and metastasis in prostate cancers. Hence, alteration of caveolin-1 expression has been proposed as a clinical marker for diagnosis and prognosis in various cancers. For precise analyses of the caveolin expression in human T cell leukemia cell lines, we measured the mRNA levels of caveolin isoforms, caveolin-1alpha, -1beta, -2, and -3 with real-time RT-PCR using external standards for each isoform. In the panel of human T cell leukemia cell lines tested, four cell lines expressed caveolin-1alpha, -1beta and -2, but not -3, which was consistent with the protein levels. The expression profiles in most cell lines are caveolin-1alpha > caveolin-1beta > caveolin-2. Two cell lines did not express either of the caveolin mRNAs. Methylation analyses for the CpG sites in the promoter region of a positive and a negative cell line did not show a clear correlation with the expression status, suggesting that mechanisms other than CpG methylation are involved in the regulation of caveolin-1alpha expression in human T cell leukemia cell lines.

Up-regulation of genes for oxidative phosphorylation and protein turnover in diabetic mouse retina.

Diabetic retinopathy is one of the most frequent complications of diabetes and is a leading cause of vision loss in adulthood. To better understand the molecular pathophysiology of diabetic retinopathy, we performed comprehensive gene expression analysis of the mouse retina under diabetic conditions with an in-house cDNA microarray system that was designed to be suitable for the small amount of RNA available from a single mouse retina. Diabetes was induced in male C57BL/6 mice by an intraperitoneal injection of streptozotocin, and the changes in retinal mRNA levels were examined in three pairs of diabetic and age-matched control mice at 1 and 3 months after the injection of streptozotocin. Northern blot analysis with amplified total cRNA confirmed the increase in mRNA levels of several selected genes. Most of the significantly up-regulated genes could be classified into two functional categories: oxidative phosphorylation and protein turnover. All mitochondrial DNA-encoded and most of the nuclear DNA-encoded genes for oxidative phosphorylation were up-regulated in the diabetic retina. This was in sharp contrast with a previous report of a down-regulation of these genes in skeletal muscles of streptozotocin-induced diabetic mice and type 2 diabetic humans. Genes for protein synthesis and ubiquitin were also up-regulated in the diabetic retina, suggesting the increase in turnover rates for at least a part of the protein population. Taken together, the diabetic retina appears to be in a state activated for intermediary metabolism, presumably because of an increase in insulin-independent glucose influx. These results provide insights into possible preventive and therapeutic intervention of diabetic retinopathy.

Activation of genes for growth factor and cytokine pathways late in chondrogenic differentiation of ATDC5 cells.

The mouse embryonal carcinoma cell line ATDC5 provides an excellent model system for chondrogenesis in vitro. To understand better the molecular mechanisms of endochondral bone formation, we investigated gene expression profiles during the differentiation course of ATDC5 cells, using an in-house microarray harboring full-length-enriched cDNAs. For 28 days following chondrogenic induction, 507 genes were up- or down-regulated at least 1.5-fold. These genes were classified into five clusters based on their expression patterns. Genes for growth factor and cytokine pathways were significantly enriched in the cluster characterized by increases in expression during late stages of chondrocyte differentiation. mRNAs for decorin and osteoglycin, which have been shown to bind to transforming growth factors-beta and bone morphogenetic proteins, respectively, were found in this cluster and were detected in hypertrophic chondrocytes of developing mouse bones by in situ hybridization analysis. Taken together with assigned functions of individual genes in the cluster, interdigitated interaction between a number of intercellular signaling molecules is likely to take place in the late chondrogenic stage for autocrine and paracrine regulation among chondrocytes, as well as for chemoattraction and stimulation of progenitor cells of other lineages.

PKC pathway and ERK/MAPK pathway are required for induction of cyclin D1 and p21Waf1 during 12-o-tetradecanoylphorbol 13-acetate-induced differentiation of myeloleukemia cells.

Treatment of human promyelocytic leukemia cell HL60 with 12-o-tetradecanoylphorbol 13-acetate (TPA) induces growth arrest, differentiation towards the monocyte/macrophage lineage, and expression of cell cycle-regulating genes cyclin D1 and p21Waf1. First, we demonstrated that p21Waf1 expression was increased by TPA in other leukemia cell lines also, including THP-1, U937, and KG-1, which differentiate into monocytes/macrophages by TPA. Secondly, we demonstrated the signal transduction pathways of cyclin D1 and p21Waf1 expressions in TPA-treated HL60 cells. Induction of cyclin D1 expression in TPA-treated HL60 cells was inhibited with protein kinase C (PKC) inhibitor bisindolylmaleimide I and mitogen activated protein kinase kinase (MEK) inhibitor PD98059. Induction of p21Waf1 expression in TPA-treated HL60 cells was inhibited with PKC inhibitor bisindolylmaleimide I and Gö6976, MEK inhibitor PD98059, and p38 mitogen-actibated protein kinase (MAPK) inhibitor SB202190. Thus, cyclin D1 and p21Waf1 expressions are considered to be induced via PKC and extracellular signal-regulated kinase/mitogen-activated protein kinase (MAPK/ERK) pathways in TPA-treated HL60 cells. The upregulation of p21Waf1 seems to play a critical role in TPA-induced cell differentiation by suppressing cyclin dependent kinase activity , while the upregulation of cyclin D1 seems to be compensated by p21Waf1.

Neuroprotective and neurotoxic effects of cyclosporine A on transient focal ischemia in mdr1a knockout mice.

The proper dose of cyclosporine A as a neuroprotective agent was investigated using the middle cerebral artery occlusion model of mdr1a knockout mice. After a 30-min occlusion period, reperfusion was performed and the vehicle or cyclosporine A (1 mg/kg or 10 mg/kg x 2) was intraperitoneally administered to each animal model group. Forty eight hours after reperfusion, infarction volume in the 1 mg/kg cyclosporine A group was significantly less than that seen in the vehicle group, although, in the high dose cyclosporine A group, infarction volumes were significantly higher than those seen in the vehicle group. These results demonstrate that cyclosporine A shows not only anti-ischemic effects, but also neurotoxic effects depending on the dosage penetrating into the brain.

Asymmetric activation to the context-dependent right in the right inferior frontal region.

In human spatial recognition, right and left are not recognized symmetrically. Although there have been many studies on the hemispheric asymmetry of the human brain, asymmetries in high-level recognition (such as independence from input or output hemisphere) have not been studied extensively. We found that the human brain recognizes right and left asymmetrically in high-level recognition. Experiments were performed in which participants crossed their hands and were required to judge the side of a tactile stimulus on the index finger in two different contexts: 'which hand was touched' or 'on which side of the space the touched hand was located'. The right inferior frontal region was significantly more activated by the 'contextually defined right' stimulus (right-hand stimulation in the 'which hand' context and right-space stimulation in the 'which space' context) than by the 'contextually defined left' stimulus. However, no activation that was more activated by the 'contextually defined left' than by the 'contextually defined right' was found. This asymmetric activation suggests that 'right' is the more outstanding side for human spatial recognition.

Neural mechanisms of spatial stimulus-response compatibility: the effect of crossed-hand position.

Previous psychological experiments have indicated the existence of a visual-proprioceptive interaction in spatial stimulus-response compatibility (SSRC) tasks, but there is little specific information on the neural basis of such interaction in humans. Using functional magnetic resonance imaging (fMRI), we compared the neural activity associated with two different aspects of spatial coding: the coding of the "internal" spatial position of motor-response effectors (i.e., the position of body parts) as obtained through proprioception, and the coding of "external" positions, i.e., the positions of visual stimuli. A 2 x 2 factorial design was used to investigate the spatial compatibility (incompatible versus compatible) between a visual stimulus and hand position (crossed versus uncrossed). The subjects were instructed to respond to stimuli presented to the right or left visual field with either the ipsilateral (compatible condition) or the contralateral hand (incompatible condition). The incompatible condition produced stronger activation in the bilateral superior parietal lobule, inferior parietal lobule, and bilateral superior frontal gyrus than the compatible condition. The crossed-hand condition produced stronger activation in the bilateral precentral gyrus, superior frontal gyrus, superior parietal lobule, and superior temporal gyrus than the uncrossed-hand condition. These results suggest that activity in the frontal-parietal regions is related to two functions: (1) representation of the visual stimulus-motor response spatial configuration in an SSRC task, and (2) integration between external visual and internal proprioceptive sensory information. The activation in the superior temporal gyrus was not affected by the visual stimulus-motor response spatial configuration in an SSRC task; rather, it was affected by the crossed-hand posture. Thus, it seems to be related to representing internal proprioceptive sensory information necessary to carry out motor actions.

Protein kinase C-gamma and calcium/calmodulin-dependent protein kinase II-alpha are persistently translocated to cell membranes of the rat brain during and after middle cerebral artery occlusion.

The levels of protein kinase C-gamma (PKC-gamma ) and the calcium/calmodulin-dependent kinase II-alpha (CaMKII-alpha) were measured in crude synaptosomal (P2), particulate (P3), and cytosolic (S3) fractions of the neocortex of rats exposed to 1-hour and 2-hour middle cerebral artery occlusion (MCAO) and 2-hour MCAO followed by 2-hour reperfusion. During MCAO, PKC levels increased in P2 and P3 in the most severe ischemic areas concomitantly with a decrease in S3. In the penumbra, PKCgamma decreased in S3 without any significant increases in P2 and P3. Total PKC-gamma also decreased in the penumbra but not in the ischemic core, suggesting that the protein is degraded by an energy-dependent mechanism, possibly by the 26S proteasome. The CaMKII-alpha levels increased in P2 but not P3 during ischemia and reperfusion in all ischemic regions, particularly in the ischemic core. Concomitantly, the levels in S3 decreased by 20% to 40% in the penumbra and by approximately 80% in the ischemic core. There were no changes in the total levels of CaMKII-alpha during MCAO. The authors conclude that during and after ischemia, PKC and CaMKII-alpha are translocated to the cell membranes, particularly synaptic membranes, where they may modulate cellular function, such as neurotransmission, and also affect cell survival. Drugs preventing PKC and/or CaMKII-alpha translocation may prove beneficial against ischemic cell death.