The brain selenoproteome: priorities in the hierarchy and different levels of selenium homeostasis in the brain of selenium-deficient rats

The application of radionuclides for the localization of essential trace elements in vivo and the characterization of their binding proteins is a story of intermittently made improvements of the techniques used for their detection. In this study we present the use of neutron activation analysis and different autoradiographic imaging methods including real-time digital autoradiography to reveal new insights in the hierarchy of selenium homeostasis. Selenoproteins containing the essential trace element selenium play important roles in the CNS. Although the CNS does not show the highest selenium concentration in the case of selenium-sufficient supply in comparison with other organs, it shows a high priority for selenium uptake and retention in the case of dietary selenium deficiency. To characterize the hierarchy of selenium supply in the brain, in vivo radiotracer labeling with ⁷⁵Se in rats with different selenium status was combined with autoradiographic detection of ⁷⁵Se in brain tissue sections and ⁷⁵Se-labeled selenoproteins after protein separation by two-dimensional gel electrophoresis. This study demonstrates significant differences in the uptake of ⁷⁵Se into the brain of rats with different selenium status. A brain region-specific uptake pattern of the radiotracer ⁷⁵Se in selenium-deficient rats could be revealed and the CSF was identified as a key part of the brain selenium homeostasis.     

Astrocytes: regulation of brain homeostasis via apolipoprotein E

Astrocytes are derived from the ventricular and subventricular zones of the neural plate, though there is controversy over their derivation from astrocyte-specific precursor cells or radial glia intermediates. Astrocytes are the most abundant cell type in the brain and contribute to brain homeostasis in several ways, including buffering of extracellular K+, regulating neurotransmitter release, forming the blood-brain barrier (BBB), releasing growth factors, and regulating the brain immune response. In addition, astrocytes have been shown to release apolipoprotein E (ApoE), which has been shown to regulate neurotransmission, growth factor release, and immune responses. Due to the diverse functions of astrocytes, they may play a role in a variety of diseases such as hepatic encephalopathy, multiple sclerosis, epilepsy, and age-related diseases including Alzheimer's disease and Parkinson's disease. This review highlights many of the diverse roles played by astrocytes in regulating brain homeostasis and discusses their potential role in a variety of disorders.     

Arterial-venous network formation during brain vascularization involves hemodynamic regulation of chemokine signaling

During angiogenic sprouting, newly forming blood vessels need to connect to the existing vasculature in order to establish a functional circulatory loop. Previous studies have implicated genetic pathways, such as VEGF and Notch signaling, in controlling angiogenesis. We show here that both pathways similarly act during vascularization of the zebrafish central nervous system. In addition, we find that chemokine signaling specifically controls arterial-venous network formation in the brain. Zebrafish mutants for the chemokine receptor cxcr4a or its ligand cxcl12b establish a decreased number of arterial-venous connections, leading to the formation of an unperfused and interconnected blood vessel network. We further find that expression of cxcr4a in newly forming brain capillaries is negatively regulated by blood flow. Accordingly, unperfused vessels continue to express cxcr4a, whereas connection of these vessels to the arterial circulation leads to rapid downregulation of cxcr4a expression and loss of angiogenic characteristics in endothelial cells, such as filopodia formation. Together, our findings indicate that hemodynamics, in addition to genetic pathways, influence vascular morphogenesis by regulating the expression of a proangiogenic factor that is necessary for the correct pathfinding of sprouting brain capillaries.     

Exogenous selenium in the brain

Summary Transcardial perfusion or intraperitoneal injections with sodium selenite result in the creation of selenium bonds that can be visualized by physical development. The present paper describes how these catalytic bonds are made visible in the tissues by surrounding them with shells of metallic silver. Based on experiments with chelating agents, the possibility that selenium-metal bonds are the catalysts is discussed. In the brain, the selenium pattern is delicate and highly laminated, the grains of silver being orderly arranged corresponding with the neuropil morphology. The precipitate is most densely packed in cortical regions. The difference in staining intensity seen in different regions of the CNS reflects the density of selenium reactive terminals. The visualized selenium bonds are predominantly located within boutons, and examination in the electron microscope reveals accumulation in the presynaptic regions. In a few places precipitates can also be found in axons, but have not been observed in perikarya or dendrites. The only non-neuronal locations of selenium were sparsely scattered, astrocyte-like neuroglia, predominantly found in the cerebellum and the hypothalamus; infrequently a few blood vessels were also stained. Sections from kidney and liver are presented as examples of localizations outside the CNS of exogenous selenium.     

Dietary selenium regulation of transcript abundance of selenoprotein N and selenoprotein W in chicken muscle tissues

Selenium (Se), selenoprotein N (SelN) and selenoprotein W (SelW) play a crucial role in muscle disorders. Se status highly regulates selenoprotein mRNA levels. However, few attempts have been performed on the effect of dietary Se supplementation on muscle SelN and SelW mRNA levels in birds. To investigate the effects of Se on the regulation of SelN and SelW mRNA levels in muscle tissues, one-day-old male chickens were fed either a commercial diet or a Se-supplemented diet containing 1.0, 2.0, 3.0 or 5.0 mg/kg sodium selenite for 90 days. Muscle tissues (breast, flight, thigh, shank and cardiac muscles) were collected and examined for Se content and mRNA levels of SelN and SelW. Moreover, Selenophosphate synthetase-1 (SPS-1) and selenocysteine-synthase (SecS) mRNA levels were analyzed. Significant increases in SelN mRNA levels were obtained in breast, thigh and shank muscles treated with Se, with maximal effects at 3.0 mg Se/kg diet, but 2.0 mg Se/kg diet resulted in peak levels of Sel N mRNA in flight muscles. Changes in SelW mRNA abundance in thigh and shank muscles increased in response to Se supply. After reaching a maximal level, higher Se supplementation led to a reduction in both SelN and SelW mRNAs. However, SelN and SelW mRNA levels displayed a different expression pattern in different skeletal and cardiac muscles. Thus, it suggested that skeletal and cardiac muscles SelN and SelW mRNA levels were highly regulated by Se supplementation and different muscle tissues showed differential sensitivity. Moreover, Se supplementation also regulated the levels of SPS1 and SecS mRNAs. The mRNA levels of SPS1 and SecS were enhanced in the Se supplemented groups. These data indicate that Se regulates the expression of SelN and SelW gene and affect the mRNA levels of SecS and SPS1.     

Cell cycle regulation of the E2F transcription factor involves an interaction with cyclin A.

We have examined E2F binding activity in extracts of synchronized NIH 3T3 cells. During the G0 to G1 transition, there is a marked increase in the level of active E2F. Subsequently, there are changes in the nature of E2F-containing complexes. A G1-specific complex increases in abundance, disappears, and is then replaced by another complex as S phase begins. Analysis of extracts of thymidine-blocked cells confirms that the complexes are cell cycle regulated. We also show that the cyclin A protein is a component of the S phase complex. Each complex can be dissociated by the adenovirus E1A 12S product, releasing free E2F. The release of E2F from the cyclin A complex coincides with the stimulation of an E2F-dependent promoter. We suggest that these interactions control the activity of E2F and that disruption of the complexes by E1A contributes to a loss of cellular proliferation control.     

Isoproterenol and cAMP regulation of the human brain natriuretic peptide gene involves Src and Rac

Brain natriuretic peptide (BNP) gene expression and chronic activation of the sympathetic nervous system are characteristics of the development of heart failure. We studied the role of the beta-adrenergic signaling pathway in regulation of the human BNP (hBNP) promoter. An hBNP promoter (-1818 to +100) coupled to a luciferase reporter gene was transferred into neonatal cardiac myocytes, and luciferase activity was measured as an index of promoter activity. Isoproterenol (ISO), forskolin, and cAMP stimulated the promoter, and the beta(2)-antagonist ICI 118,551 abrogated the effect of ISO. In contrast, the protein kinase A (PKA) inhibitor H-89 failed to block the action of cAMP and ISO. Pertussis toxin (PT), which inactivates Galpha(i), inhibited ISO- and cAMP-stimulated hBNP promoter activity. The Src tyrosine kinase inhibitor PP1 and a dominant-negative mutant of the small G protein Rac also abolished the effect of ISO and cAMP. Finally, we studied the involvement of M-CAT-like binding sites in basal and inducible regulation of the hBNP promoter. Mutation of these elements decreased basal and cAMP-induced activity. These data suggest that beta-adrenergic regulation of hBNP is PKA independent, involves a Galpha(i)-activated pathway, and targets regulatory elements in the proximal BNP promoter.     

The selenium intake of the female chicken influences the selenium status of her progeny

The primary purpose of this study is to determine the extent to which the effects of dietary supplementation of the female chicken with selenium (Se) continue into the next generation. An additional aim is to compare the relative effectiveness of pre-hatch (from the hen's diet) with that of post-hatch (from the progeny's diet) supplementation with Se on the Se status of the chick during the first 4 weeks of post-hatch life. Hens were maintained on control or Se-supplemented diets, respectively containing 0.027 and 0.419 μg Se/g of feed. The high-Se diet elevated the Se content of the hens' eggs by 7.1-fold. At hatch, the concentrations of Se in the liver, breast muscle and whole blood of the chicks originating from the high-Se parents were, respectively, 5.4-, 4.3- and 7.7-fold higher than the values in the chicks of the low-Se parents. When the offspring from the two parental groups were both maintained on the low-Se progeny diet, the tissue Se concentrations in chicks originating from the high-Se hens remained significantly higher for 3–4 weeks after hatching, compared with the values in chicks from the low-Se hens. Similarly, tissue glutathione peroxidase activity remained significantly higher in chicks from the high-Se hens for 2–4 weeks post-hatch. Thus, the effects of maternal Se supplementation persist in the progeny for several weeks after hatching. However, when chicks hatching from low-Se eggs were placed on a high Se diet, their tissue Se concentrations at 7 days of age were markedly higher than the values in chicks from high-Se eggs placed on the low-Se diet.     

Evolutionary conservation of KLF transcription factors and functional conservation of human gamma-globin gene regulation in chicken

The Krüppel-like factors (KLFs) are a family of Cys2His2 zinc-finger DNA binding proteins with homology to Drosophila Krüppel. KLFs can bind to CACCC elements, which are important in controlling developmental programs. The CACCC promoter element is critical for the developmental regulation of the human gamma-globin gene. In the present study, chicken homologues of the human KLF2, 3, 4, 5, 9, 11, 12, 13, and 15 genes were identified. Phylogenetic analysis confirms that these genes are more closely related to their human homologues than they are to other chicken KLFs. This work also represents the first systematic study of the expression patterns of KLFs during erythroid development. In addition, transient transfections of human globin constructs into 5-day (primitive) chicken red blood cells show that human gamma-globin expression is regulated via its CACCC promoter element. This indicates that a CACCC-binding factor(s) important for gamma-globin expression functions in 5-day chicken red cells.     

Calcium-dependent regulation of glucose homeostasis in the liver

A major role of the liver is to integrate multiple signals to maintain normal blood glucose levels. The balance between glucose storage and mobilization is primarily regulated by the counteracting effects of insulin and glucagon. However, numerous signals converge in the liver to ensure energy demand matches the physiological status of the organism. Many circulating hormones regulate glycogenolysis, gluconeogenesis and mitochondrial metabolism by calcium-dependent signaling mechanisms that manifest as cytosolic Ca²⁺ oscillations. Stimulus-strength is encoded in the Ca²⁺ oscillation frequency, and also by the range of intercellular Ca²⁺ wave propagation in the intact liver. In this article, we describe how Ca²⁺ oscillations and waves can regulate glucose output and oxidative metabolism in the intact liver; how multiple stimuli are decoded though Ca²⁺ signaling at the organ level, and the implications of Ca²⁺ signal dysregulation in diseases such as metabolic syndrome and non-alcoholic fatty liver disease.     

Cholesterol metabolism and homeostasis in the brain

Cholesterol is an essential component for neuronal physiology not only during development stage but also in the adult life. Cholesterol metabolism in brain is independent from that in peripheral tissues due to bloodbrain barrier. The content of cholesterol in brain must be accurately maintained in order to keep brain function well. Defects in brain cholesterol metabolism has been shown to be implicated in neurodegenerative diseases, such as Alzheimer’s disease (AD), Huntington’s disease (HD), Parkinson’s disease (PD), and some cognitive deficits typical of the old age. The brain contains large amount of cholesterol, but the cholesterol metabolism and its complex homeostasis regulation are currently poorly understood. This review will seek to integrate current knowledge about the brain cholesterol metabolism with molecular mechanisms.