Search

Brain aging is characterized by progressive loss of plasma membrane calcium pump (PMCA) and its activator – calmodulin (CaM), but the mechanism of this phenomenon remains unresolved. CaM encoded by three genes Calm1 , Calm2 , Calm3 , works to translate Ca 2 + signal into changes in frequently opposite cellular activities. This unique function allows CaM to affect gene expression via stimulation of calcineurin (CaN) and its downstream target – nuclear factor of activated T-cells (NFAT) and to terminate Ca 2 + signal by stimulation of its extrusion. PMCA, which exists in four isoforms PMCA1–4, may in turn shape the pattern of Ca 2 + transients and control CaN activity by its direct binding. Therefore, the interplay between PMCA, CaM and CaN/NFAT is highly plausible. To verify that, we used differentiated PC12 cells with reduced expression of PMCA2 or PMCA3 to mimic the potential changes in aged brain. Manipulation in PMCAs level decreased CaM protein in PMCA2 or PMCA3-reduced lines that was accompanied by down-regulation of Calm1 and Calm2 in both lines, but Calm3 only in PMCA2-reduced cells. Further studies showed substantially higher NFATc2 nuclear accumulation and increased NFAT transcriptional activity. Blocking of CaN/NFAT signalling resulted in almost full CaM recovery, mainly due to up-regulation of Calm2 and Calm3 genes. Moreover, higher occupancy of Calm2 and Calm3 promoters by NFATc2 and increased expression of these genes in response to NFATc2 silencing were demonstrated in PMCA2 and PMCA3-reduced lines. Our results indicate that decrease in CaM level in response to PMCAs downregulation can be driven by CaN/NFAT pathway. [ABSTRACT FROM AUTHOR]

Abstract: Changes in PMCA2 and PMCA3 expression during neuronal development are tightly linked to structural and functional modifications in Ca2+ handling machinery. Using antisense strategy we obtained stably transfected PC12 lines with reduced level of PMCA2 or PMCA3, which were then subjected to dibutyryl-cAMP differentiation. Reduced level of neuron-specific PMCAs led to acceleration of differentiation and formation of longer neurites than in control PC12 line. Treatment with dibutyryl-cAMP was associated with retraction of growth cones and intensified formation of varicosities. In PMCA2-reduced cells development of apoptosis and DNA laddering were detected. Higher amounts of constitutive isoforms PMCA1 and PMCA4, their putative extended location to gaps left after partial removal of PMCA2 or PMCA3, together with increased SERCA may indicate the induction of compensatory mechanism in modified cells. Functional studies showed altered expression of certain types of VDCCs in PMCA-reduced cells, which correlated with their higher contribution to Ca2+ influx. The cell response to PMCAs suppression suggests the interplay between transcription level of two opposite calcium-transporting systems i.e. voltage- and store depletion-activated channels facilitating Ca2+ influx and calcium pumps responsible for Ca2+ clearance, as well highlights the role of both neuron-specific PMCA isoforms in the control of PC12 cells differentiation. [Copyright &y& Elsevier]

Objectives: Perinatal asphyxia represents a major cause of acute brain impairment and mortality in neonates. To develop the effective therapies able to reduce post-asphyxial damages, the understanding of biochemical processes accompanying asphyxia appears to be of the great relevance.Design and methods: The activities of protein kinases A and C, and tyrosine kinases in erythrocyte membranes of healthy and asphyxiated neonatals were compared. Using monoclonal antibodies the band 3 presence and its phosphotyrosine levels were assayed.Results: In asphyxiated erythrocyte membranes the activities of PKA and tyrosine kinases increased, whereas the activity of PKC was reduced in relation to healthy newborns. Under asphyxia the band 3 has been overphosphorylated; however, its amount decreased.Conclusion: These findings may provide some evidence for a potential role of asphyxia in disturbance of phosphorylation processes in erythrocytes, as reflected by altered protein kinases activities. The diminished band 3 presence may be partially responsible for the impairment of erythrocyte function. [Copyright &y& Elsevier]

Although first mentions about calcium disturbances in psychiatric diseases appeared more than 30 years ago, the most recent genomic and proteomic findings confirmed a significant role of Ca 2+ and Ca 2+ -regulated pathways in development of neuropathological processes, including bipolar disorder and schizophrenia. Moreover, last decades have shown that due to multifactorial nature of both diseases, impairment in neuronal calcium homeostasis may depend not only on disturbed Ca 2+ entry system, but also on altered extrusion system. A pivotal role in Ca 2+ clearance mechanism is played by plasma membrane Ca 2+ -ATPase (PMCA), the enzyme responsible for returning the elevated levels of cytosolic Ca 2+ back to the resting state. In this paper we summarize the current knowledge about the role of PMCA in bipolar disorder and schizophrenia pathologies, as well as the contribution of several proteins that by interaction with PMCA modify signal transduction mechanisms. [ABSTRACT FROM AUTHOR]

A growing body of evidence indicates that clinical use of ketamine as a promising antidepressant can be accompanied by psychotic-like side effects. Although, the generation of such effects is thought to be attributed to dysfunction of prefrontal GABAergic interneurons, the mechanism underlying ketamine's propsychotic-like action is not fully understood. Due to wide spectrum of behavioral abnormalities, it is hypothesized that ketamine action is not limited to only cortical GABA metabolism but may also involve alterations in other functional brain areas. To test it, we treated rats with ketamine (30 mg/kg, i.p.) for 5 days, and next we analyzed GABA metabolizing enzymes in cortex, cerebellum, hippocampus and striatum. Our results demonstrated that diminished GAD67 expression in cortex, cerebellum (by ∼60%) and in hippocampus (by ∼40%) correlated with lowered protein level in these areas. The expression of GAD65 isoform decreased by ∼45% in striatum, but pronounced increase by ∼90% was observed in hippocampus. Consecutively, reduction in glutamate decarboxylase activity and GABA concentration were detected in cortex, cerebellum and striatum, but not in hippocampus. Ketamine administration decreased GABA transaminase protein in cortex and striatum (by ∼50% and 30%, respectively), which was reflected in diminished activity of the enzyme. Also, a significant drop in succinic semialdehyde dehydrogenase activity in cortex, cerebellum and striatum was present. These data suggest a reduced utilization of GABA for energetic purposes. In addition, we observed synaptic GABA release to be reduced by ∼30% from striatal terminals. It correlated with lowered KCl-induced Ca 2+ influx and decreased amount of L-type voltage-dependent calcium channel. Our results indicate that unique changes in GABA metabolism triggered by chronic ketamine treatment in functionally distinct brain regions may be involved in propsychotic-like effects of this drug. [ABSTRACT FROM AUTHOR]

Among polychlorinated naphthalenes (PCNs), listed by the Stockholm convention as Persistent Organic Pollutants (POPs), hexachloronaphthalenes are considered the most toxic and raise the highest concern. Of these, 1,2,3,5,6,7-hexachloronaphthalanene (PCN67) is considered the main congener affecting human health due to its hepatotoxicity and its ability to disturb the reproductive, endocrine, and hematological systems. It is also prevalent in human serum/plasma, milk, and adipose tissue. However, little is known about its neurotoxicity, despite the fact that anorectic effects have been observed in workers occupationally exposed to PCNs and in animal research on PCN67. Since dopamine is involved in many aspects of food intake, the aim of this study was to confirm whether PCN67 affects dopamine synthesis in differentiated PC12 cells, a widely used model of neurosecretion. Our results show that exposure to PCN67 resulted in diminished dopamine content and release. Moreover, PCN67 also affected the expression of tyrosine hydroxylase and lowered the expression of vesicular monoamine transporter 1 (VMAT1). In addition, significantly lower expression of antioxidant enzymes, including catalase, glutathione peroxidase and copper/zinc superoxide dismutase, was observed in comparison to the vehicle. In conclusion, PCN67 appears to disturb dopaminergic transmission by altering tyrosine hydroxylation, reducing VMAT1 expression and impairing antioxidant protection. Our study provides a potential mechanism for how PCN67 may cause dopamine deficiency and contribute to neuronal death by affecting cellular antioxidant potency; however, this conclusion requires further research. [Display omitted] • Effects of PCN67 on in vitro dopaminergic secretion were studied. • PCN67 significantly inhibits biosynthesis of dopamine in PC12 cells. • PCN67 significantly decreased expression of CAT, GPx and SOD in PC12 cells. [ABSTRACT FROM AUTHOR]

Abstract: Objectives:: Magnesium sulfate has been recognized as a neuroprotective agent against hypoxia–ischemia, mainly by the protection from the excitotoxicity associated with increased glutamate concentration. However, the mechanism of MgSO4 action is not fully understood and is considerably controversial. Design and methods:: During the 2 first hours of life, the asphyxiated full-term newborns were treated intravenously with one dose of MgSO4 250 mg/kg body weight. At birth, after 6 and 48 h of life the activity of ATP-dependent enzymes in erythrocyte membranes: Mg2+-ATPase, Ca2+-ATPase, protein kinases A and C, were determined. Using monoclonal antibodies, the band 3 and its phosphotyrosine level were also assayed. Results:: The time-dependent decrease of Ca2+-ATPase activity was detected in untreated newborns, whereas MgSO4 prevented this reduction. After 48 h, protein kinases activities differed in MgSO4-treated and untreated groups. Magnesium therapy increased the amount of band 3 and diminished proteolytic degradation of this protein. Conclusion:: Our results demonstrated, for the first time, that magnesium sulfate treatment significantly altered the activities of some important enzymes in erythrocyte membrane from asphyxiated newborns. It also reduced the post-asphyxial damages of membrane compounds. These data may partly explain the molecular mechanisms of MgSO4 action in asphyxiated newborns. [Copyright &y& Elsevier]