Angiotensin receptor type 1 blockade in astrocytes decreases hypoxia-induced cytotoxicity and inflammation

The present study investigated the role of angiotensin receptors (AT-R) in the survival and inflam- matory response of astroglia upon hypoxic injury. Expo- sure of rat astroglial primary cultures (APC) to hypoxic conditions (HC) led to decreased viability of the cells and to a 3.5-fold increase in TNF-alpha release. AT-R type1 (AT1-R) antagonist losartan and its metabolite EXP3174 decrease the LDH release (by 36 ± 9%; 45 ± 6%) from APC under HC. Losartan diminished TNF-alpha release (by 40 ± 15%) and the number of TUNEL-cells by 204 ± 38% under HC, alone and together with angiotensin II (ATII), while EXP3174 was dependent on ATII for its effect on TNF-alpha. The AT2-R antagonist, PD123.319, did not influence the release of LDH and TNF-alpha under normoxic (NC) and HC. These data suggest that AT1-R may decrease the susceptibility of astrocytes to hypoxic injury and their propensity to release TNF-alpha. AT1-R antagonists may therefore be of therapeutic value during hypoxia-associated neurodegeneration

Primary cultures as a model for studying ependymal functions: glycogen metabolism in ependymal cells

Ependymal cells form a single-layered, ciliated epithelium at the interface between the cerebrospinal fluid and the brain parenchyma. Although their morphology has been studied in detail, ependymal functions remain largely speculative. We have established and characterized a previously described cell culture model to investigate ependymal glycogen metabolism. During growth in minimal medium lacking many non-essential amino acids including L-glutamate, but containing glucose at physiological concentration, the cells contained negligible amounts of glycogen (7 +/- 3 nmol glucosyl residues/mg protein) despite the presence of insulin. However, during a period of 24 h, the cells accumulated glycogen to very high levels after transferal to a medium containing insulin, glucose at a 5-fold higher concentration. and all proteinogenic amino acids except L-asparagine and L-serine (990 +/- 112 nmol glucosyl residues/mg protein). omission of insulin resulted in a 50%, reduction in glycogen accumulation. Upon glucose deprivation, glycogen was degraded with a half-life of 21 min. The ependymal primary cultures contained 80 +/- 5 mU glycogen phosphorylase (Pho)/mg protein and stained positively with antibodies raised against this enzyme. Astroglial cultures built up less glycogen and had less Pho activity under identical conditions. Ependymal glycogen was mobilized by noradrenaline and serotonin. Our results indicate that ependymal cells maintain glycogen as a functional energy store, subject to rapid turnover dependent on the availability of energy substrates and the presence of appropriate signal molecules. Thus ependymocytes appear to be active players in the multitude of processes resulting in normal brain function, and ependymal primary cultures are suggested as a suitable model for studying the role of ependymal cells in these processes. (C) 2001 Elsevier Science B.V. All rights reserved

Comparison of lactate transport in astroglial cells and monocarboxylate transporter 1 (MCT 1) expressing Xenopus laevis oocytes. Expression of two different monocarboxylate transporters in astroglial cells and neurons

The transport of lactate is an essential part of the concept of metabolic coupling between neurons and glia. Lactate transport in primary cultures of astroglial cells was shown to be mediated by a single saturable transport system with a Km value for lactate of 7.7 mM and a Vmax value of 250 nmol/(min x mg of protein). Transport was inhibited by a variety of monocarboxylates and by compounds known to inhibit monocarboxylate transport in other cell types, such as alpha-cyano-4-hydroxycinnamate and p-chloromercurbenzenesulfonate. Using reverse transcriptase-polymerase chain reaction and Northern blotting, the presence of mRNA coding for the monocarboxylate transporter 1 (MCT1) was demonstrated in primary cultures of astroglial cells. In contrast, neuron-rich primary cultures were found to contain the mRNA coding for the monocarboxylate transporter 2 (MCT2). MCT1 was cloned and expressed in Xenopus laevis oocytes. Comparison of lactate transport in MCT1 expressing oocytes with lactate transport in glial cells revealed that MCT1 can account for all characteristics of lactate transport in glial cells. These data provide further molecular support for the existence of a lactate shuttle between astrocytes and neurons

Comparison of lactate transport in astroglial cells and monocarboxylate transporter 1 (MCT 1) expressing Xenopus laevis oocytes: Expression of two different monocarboxylate transporters in astroglial cells and neurons

The transport of lactate is an essential part of the concept of metabolic coupling between neurons and glia. Lactate transport in primary cultures of astroglial cells was shown to be mediated by a single saturable transport system with a Km value for lactate of 7.7 mM and a Vmax value of 250 nmol/(min x mg of protein). Transport was inhibited by a variety of monocarboxylates and by compounds known to inhibit monocarboxylate transport in other cell types, such as alpha-cyano-4-hydroxycinnamate and p-chloromercurbenzenesulfonate. Using reverse transcriptase-polymerase chain reaction and Northern blotting, the presence of mRNA coding for the monocarboxylate transporter 1 (MCT1) was demonstrated in primary cultures of astroglial cells. In contrast, neuron-rich primary cultures were found to contain the mRNA coding for the monocarboxylate transporter 2 (MCT2). MCT1 was cloned and expressed in Xenopus laevis oocytes. Comparison of lactate transport in MCT1 expressing oocytes with lactate transport in glial cells revealed that MCT1 can account for all characteristics of lactate transport in glial cells. These data provide further molecular support for the existence of a lactate shuttle between astrocytes and neurons

Biosynthesis of Wdrl6, a marker protein for kinocilia-bearing cells, starts at the time of kinocilia formation in rat, and wdr16 gene knockdown causes hydrocephalus in zebrafish

The rat ortholog of the WD40 repeat protein Wdr16 is abundantly expressed in testis and cultured ependymal cells. Low levels are found in lung and brain, respectively, while it is absent from kinocilia-free tissues. In testis and ependymal primary cultures, Wdr16 messenger RNA appears concomitantly with the messages for sperm-associated antigen 6, a kinocilia marker, and for hydin, a protein linked to ciliary function and hydrocephalus. In testis, ependyma and respiratory epithelium, the Wdr16 protein is up-regulated together with kinocilia formation. The wdr16 gene is restricted to genera in possession of kinocilia, and it is strongly conserved during evolution. The human and zebrafish proteins are identical in 62% of their aligned amino acids. On the message level, the zebrafish Wdr16 ortholog was found exclusively in kinocilia-bearing tissues by in situ hybridisation. Gene knockdown in zebrafish embryos by antisense morpholino injection resulted in severe hydrocephalus formation with unaltered ependymal morphology or ciliary beat. Wdr16 can be considered a differentiation marker of kinocilia-bearing cells. In the brain, it appears to be functionally related to water homeostasis or osmoregulation

The Aiguille du Midi (Mont Blanc massif): a unique high-Alpine site to study bedrock permafrost

Permafrost and its change in steep high-Alpine rock walls remain insufficiently understood because of the difficulties of in situ measurements. A large proportion of permafrost studies is mainly based on modelling, with a few existing instrumented sites and a resulting lack of process understanding. Yet, a number of rockfalls that occurred in the last decade in the Alps are likely related to climatically-driven permafrost degradation, as indicated by ice in starting zones, increased air temperature, and modelling studies. Starting off in the framework of the French-Italian PERMAdataROC project and presently under development within the EU co-funded project PermaNET (Permafrost long-term monitoring network: www.permanetalpinespace. eu), our investigations at the Aiguille du Midi begin in 2005. The summit (3842 m a.s.l) is accessible from Chamonix by a cable car which was built at the end of the 1950s. Half a million tourists visit the site each year. Because of its elevation, geometry, and year-round accessibility to rock slopes of diverse aspects and to galleries, the site was chosen for: - Monitoring of the thermal regime in steep rock walls. Thermistors were installed at depths of 2, 10, 30 and 55 cm, at all aspects and with slope angles in the range 60-90° (determining e.g. the presence and influence of snow). - Measurements of high altitude climatic data (air temperature and humidity, incoming and outgoing solar radiation, wind speed and direction) perpendicular to the rockwall surface, by movable automatic weather stations. Together with the rock temperature measurements, these data (see Morra et al., poster in session CR4.1) can be used for physically-based model validation (see Pogliotti et al., oral presentation in session CR4.1) or statistical models construction of rock temperature distribution and variability in the rock walls. - Making a 3D-high-resolution DEM by long-range (rock walls) and short-range (galleries) terrestrial laser scanning. - Surveying the distribution of permafrost in the rock mass and its seasonal evolution during the year using electrical resistivity tomography and laboratory testing of temperature-resistivity behaviour of the local granite (see Krautblatter et al., poster in session CR4.1). During the next months, the following complementary instrumentation and research will be accomplished: - Numerical modelling of the 3-D distribution and evolution of temperature fields in the subsurface, based on the combination of a distributed energy balance model and a 3-D heat conduction scheme for the subsurface. - Installation of thermistor chains in shallow boreholes (up to a depth of 10 m) to monitor temperatures and to parametrize and validate rock temperature models. - Infrared thermography, using a handheld thermographic camera, to produce diachronic images of the rockwall radiative temperatures. - Numerical modelling of rock fractures and water flow in unsaturated fractures. - Monitoring of the morphological activity of the Arête des Cosmiques SE face - SE Pilastre by long-range terrestrial laserscanning. The combination of process understanding, statistical analyses and/or modelling will help to improve our understanding of where, why and how permafrost degradation in mountains occurs. Secondly, we are interested in how a reduction in the uncertainty of data, process understanding and models may contribute to our predictive skill of corresponding effects. As an illustration of this innovative research project, the poster presents some first results of the investigations at the Aiguille du Mid

Conceptual design and first results of a combined electrical resistivity and induced polarization monitoring at the Aiguille du Midi rock permafrost summit (Mont Blanc Massif, French/Italian Alps)

EGU2009-921