Long-term treadmill exercise improves spatial memory of male APPswe/PS1dE9 mice by regulation of BDNF expression and microglia activation

Increasing evidence suggests that physical activity could delay or attenuate the symptoms of Alzheimer''s disease (AD). But the underlying mechanisms are still not fully understood. To investigate the effect of long-term treadmill exercise on the spatial memory of AD mice and the possible role of β-amyloid, brain-derived neurotrophic factor (BDNF) and microglia in the effect, male APPswe/PS1dE9 AD mice aged 4 months were subjected to treadmill exercise for 5 months with 6 sessions per week and gradually increased load. A Morris water maze was used to evaluate the spatial memory. Expression levels of β-amyloid, BDNF and Iba-1 (a microglia marker) in brain tissue were detected by immunohistochemistry. Sedentary AD mice and wildtype C57BL/6J mice served as controls. The results showed that 5-month treadmill exercise significantly decreased the escape latencies (P < 0.01 on the 4th day) and improved the spatial memory of the AD mice in the water maze test. Meanwhile, treadmill exercise significantly increased the number of BDNF-positive cells and decreased the ratios of activated microglia in both the cerebral cortex and the hippocampus. However, treadmill exercise did not significantly alleviate the accumulation of β-amyloid in either the cerebral cortex or the hippocampus of the AD mice (P > 0.05). The study suggested that long-term treadmill exercise could improve the spatial memory of the male APPswe/PS1dE9 AD mice. The increase in BDNF-positive cells and decrease in activated microglia might underpin the beneficial effect.     

Biliverdin reductase-A protein levels and activity in the brains of subjects with Alzheimer disease and mild cognitive impairment

Biliverdin reductase-A is a pleiotropic enzyme involved not only in the reduction of biliverdin-IX-alpha into bilirubin-IX-alpha, but also in the regulation of glucose metabolism and cell growth secondary to its serine/threonine/tyrosine kinase activity. Together with heme oxygenase, whose metabolic role is to degrade heme into biliverdin-IX-alpha, it forms a powerful system involved in the cell stress response during neurodegenerative disorders. In this paper, an up-regulation of the biliverdin reductase-A protein levels was found in the hippocampus of the subjects with Alzheimer disease and arguably its earliest form, mild cognitive impairment. Moreover a significant reduction in the phosphorylation of serine, threonine and tyrosine residues of biliverdin reductase-A was found, and this was paralleled by a marked reduction in its reductase activity. Interestingly, the levels of both total and phosphorylated biliverdin reductase-A were unchanged as well as its enzymatic activity in the cerebella. These results demonstrated a dichotomy between biliverdin reductase-A protein levels and activity in the hippocampus of subjects affected by Alzheimer disease and mild cognitive impairment, and this effect likely is attributable to a reduction in the phosphorylation of serine, threonine and tyrosine residues of biliverdin reductase-A. Consequently, not just the increased levels of biliverdin reductase-A, but also its changed activity and phosphorylation state, should be taken into account when considering potential biomarkers for Alzheimer disease and mild cognitive impairment.     

Electrical signaling and photosynthesis: Can they co-exist together?

Mechanical irritation of trigger hairs and subsequent generation of action potentials have significant impact on photosynthesis and respiration in carnivorous Venus flytrap (Dionaea muscipula). Action potential-mediated inhibition of photosynthesis and stimulation of respiration is confined only to the trap and was not recorded in adjacent photosynthetic lamina. We showed that the main primary target of electrical signals on assimilation is in the dark enzymatic reaction of photosynthesis. Without doubt, the electrical signaling is costly, and the possible co-existence of such type of signals and photosynthesis in plant cell is discussed.Key words: action potential, carnivorous plant, Dionaea muscipula, electrical signaling, photosynthesis, respiration, Venus flytrapTrap closure of the Venus flytrap (Dionaea muscipula) is one of the fastest movements in plant kingdom. Mechanical irritation of trigger hairs protruding from upper leaf epidermis results in generation of action potential. At room temperature, two touches generate two action potentials and activate the trap snap shut in a fraction of second.¹ After the rapid movement secures the prey, struggling results in generation of further action potentials which cease to occur when the prey stops moving.² We documented that trigger hair irritation and subsequent generation of action potentials have significant effect on photosynthesis and respiration. Action potentials propagate in the trap and were not recorded in adjacent lamina (Fig. 1). This is in accordance with the observation that no changes of photosynthetic and respiration rate as well as effective quantum yield of photosystem II photochemistry were recorded in lamina. Detailed analysis of chlorophyll fluorescence kinetics revealed that the main primary target of action potentials is in the dark enzymatic reaction of photosynthesis and changes in quantum yield of primary photochemistry are just a consequence of decreased CO₂ fixation. However, electrical signals have probably also small effect on excitation energy trapping, charge stabilization and recombination reaction in photosystem II as measurements of fast chlorophyll a fluorescence transient indicates. This effect may be explained by repulsion of charges in reaction center of photosystem II.^3,4 The changes of photosynthesis upon impact of electrical signals probably have no benefit for plant and are only a negative consequences caused by the changes of the ionic environment.Open in a separate windowFigure 1Dionaea muscipula with entrapped wasp of the genus Polistes. Action potentials and rate of net assimilation at irradiance 80 µmol m⁻²s⁻ PAR (A_n) in response to 15 s mechanical trigger hair irritation (between 160–175 s) in trap (upper row) and photosynthetic lamina (lower row).These findings may have more consequences for plants in general. The electrical activity of plant cell was for the first time described by Burdon-Sanderson in 1873.⁵ Hence electrical signals do not belong exclusively to animal kingdom however they never develop the same degree of complexity as in animal nerves. Electrical signals are capable of transmitting signals more quickly over long distances when compared with chemical signals (e.g., hormones).^6,7 They are not confined only to the sensitive plants (e.g., Mimosa, Dionaea), but play also an important role in every non-sensitive plants and in both groups have significant effect on photosynthesis and respiration.^8–14 It is not surprising, that if electrical signals are costly in term of consumption of ATP and increased respiration with concurrent inhibition of photosynthesis, the same degree of complexity as in animals could not be developed. If plant growth depends on photosynthesis, this raises the question whether electrical signals and photosynthesis may co-exist together. The continuous electrical activity would inhibit the main source of energy for plants—photosynthetic assimilation. This may also explain why the plants are sessile organisms. For rapid coordinated movements, electrical activity plays an important role in animals. Unlike animals, plants usually rely on slow movements in which the role of plant hormones is indispensable. In this concept, it is not surprising that the more complex electrical activity was recorded in root transition zone—the heterotrophic part of plant body.^15,16 And this may also explain why the more evident electrical activity in the plant world has evolved in the traps of carnivorous plants like Dionaea, Aldrovanda or Drosera.^17–19 In general the traps of carnivorous plants are considered to be less efficient in photosynthesis.²⁰ Any of the action potentials produced by Drosera tentacles or Dionaea trap do not spread to photosynthetic active lamina, thus the main side of CO₂ fixation is protected.²¹ It is possible that such temporal carbon costs associated with insect trapping and retention may be outweighed by the benefits gained later from the prey—increased nitrogen concentration in the leaves stimulates photosynthetic assimilation.²² The possible ecophysiological impact of electrical signals on daily carbon gain in sensitive plants remains to be elucidated. We still do not completely understand the electrical signals in plants, and further research in this area is necessary to understand the full meaning of electrical activity in plants.     

Quantitative proteomics analysis of phosphorylated proteins in the hippocampus of Alzheimer's disease subjects

Phosphorylation on tyrosine, threonine and serine residues represents one of the most important post-translational modifications and is a key regulator of cellular signaling of multiple biological processes that require a strict control by protein kinases and protein phosphatases. Abnormal protein phosphorylation has been associated with several human diseases including Alzheimer's disease (AD). One of the characteristic hallmarks of AD is the presence of neurofibrillary tangles, composed of microtubule-associated, abnormally hyperphosphorylated tau protein. However, several others proteins showed altered phosphorylation levels in AD suggesting that deregulated phosphorylation may contribute to AD pathogenesis. Phosphoproteomics has recently gained attention as a valuable approach to analyze protein phosphorylation, both in a quantitative and a qualitative way. We used the fluorescent phosphospecific Pro-Q Diamond dye to identify proteins that showed alterations in their overall phosphorylation in the hippocampus of AD vs. control (CTR) subjects. Significant changes were found for 17 proteins involved in crucial neuronal process such as energy metabolism or signal transduction. These phosphoproteome data may provide new clues to better understand molecular pathways that are deregulated in the pathogenesis and progression of AD.     

Caspase-3 inhibits the growth of breast cancer cells independent of protease activity

In this study, the MCF-7 breast cancer cells that lack caspase-3 were transfected with a wild type (WT) or mutant caspase-3 cDNA. Expression of the WT, but not of the mutant, caspase-3 was associated with increased caspase activity and susceptibility to staurosporine (STS)-induced apoptosis. Both derivatives displayed inhibition of cell growth compared with vector control cells. Growth inhibition was associated with increased expression of the cyclin dependent kinase (CDK) inhibitor p27Kip1 in the WT, but not in the mutant caspase-3 expressing cells. Cyclin D1 expression level was not affected by caspase-3 expression. Phosphorylation of the Akt protein was decreased in both WT and mutant caspase transfected cells, although Akt expression level remained unchanged. These results suggest that caspase-3 might have biological functions independent of its protease activity and that its loss might contribute to tumor development by increasing the growth potential of cancer cells.     

MDR-like ABC transporter AtPGP4 is involved in auxin-mediated lateral root and root hair development

Previous data have suggested an involvement of MDR/PGP-like ABC transporters in transport of the plant hormone auxin and, recently, AtPGP1 has been demonstrated to catalyze the primary active export of auxin. Here we show that related isoform AtPGP4 is expressed predominantly during early root development. AtPGP4 loss-of-function plants reveal enhanced lateral root initiation and root hair lengths both known to be under the control of auxin. Further, atpgp4 plants show altered sensitivities toward auxin and the auxin transport inhibitor, NPA. Finally, mutant roots reveal elevated free auxin levels and reduced auxin transport capacities. These results together with yeast growth assays suggest a direct involvement of AtPGP4 in auxin transport processes controlling lateral root and root hair development.     

Accumulation of long-chain acylcarnitine and 3-hydroxy acylcarnitine molecular species in diabetic myocardium: identification of alterations in mitochondrial fatty acid processing in diabetic myocardium by shotgun lipidomics

Diabetic cardiomyopathy is the result of maladaptive changes in energy homeostasis. However, the biochemical mechanisms underlying dysfunctional lipid metabolism in diabetic myocardium are incompletely understood. Herein, we exploit shotgun lipidomics to demonstrate a 4-fold increase in acylcarnitines in diabetic myocardium, which was reversible upon insulin treatment. Analysis of acylcarnitine molecular species in myocardium unexpectedly identified acylcarnitine molecular species containing a mass shift of 16 amu in comparison to the anticipated molecular species. Synthesis of 3-hydroxy acylcarnitine identified the natural products as the 3-hydroxylated acylcarnitines through comparisons of diagnostic fragmentation patterns of synthetic and naturally occurring constituents using tandem mass spectrometry. Diabetes induced an increase of both calcium-independent phospholipase A(2) (iPLA(2)) mRNA and iPLA(2) activity in rat myocardium. Cardiac ischemia in myocardium genetically engineered to overexpress iPLA(2) dramatically increased the amount of acylcarnitine present in myocardium. Moreover, mechanism-based inactivation of iPLA(2) in either wild-type or transgenic myocardium ablated a substantial portion of the acylcarnitine increase. Collectively, these results identify discrete insulin remediable abnormalities in mitochondrial fatty acid processing in diabetic myocardium and identify iPLA(2) as an important enzymatic contributor to the pool of fatty acids that can be used for acylcarnitine synthesis and energy production in myocardium.