Slice cultures were allowed to settle for 7—10 days before being used for experimentation to allow AVV-mediated expression to be fully established. Primary cultures of Wistar rat pups p2 cerebral astrocytes adapted from 62 transduced with AVV. Ratio of channel 1: channel 2 is indicative of a pH change. Astrocytes were imaged in time-lapse mode 0. In these experiments a red-coloured dye was added to the pipette usually SNARF-5 to confirm successful dialysis of the patched neuron.
For further details see ref. Optogenetic stimulation was applied to the LC through an optical fiber 0.
CFEs were retracted from the tissue at the end of experiments and calibrated against NE 0. Independently of the electrode sensitivity, we can expect variability in the number of local release sites around any recording position, thus resulting in variation of measured NE volume transmission between recording sites or slices. Therefore data sets including the relevant and comparable control responses were pooled for evaluation of effects. Adequate anaesthesia was ensured by maintaining stable levels of arterial blood pressure and heart rate.
The femoral artery and vein were cannulated for measurement of arterial blood pressure and administration of anesthetic, respectively. The body temperature was maintained with a servo-controlled heating pad at For EEG recordings, two small holes were drilled on the left side of the skull above the somatosensory cortex. Stainless steel electrodes were placed in the contact with the cortical surface and secured in place with dental cement.
Drugs microinjected from pipettes in small volumes are subject to very rapid dilution and therefore activate a much larger area than the actual volume of injection. In addition, metabolites such as glutamate of L -lactate are taken up by the cells and also eliminated from the tissue into the bloodstream. On the basis of the effective concentrations of L -lactate in vitro established in this work, L -lactate could be applied at concentrations much higher than those of L -glutamate because L -glutamate receptors have high affinity, while the putative L -lactate receptor must have low affinity.
Nevertheless we decided not to increase it further to avoid possible complications due to an osmotic effect. The sites of microinjections were marked by injection of fluorescent beads, identified Post hoc histologically and mapped using a stereotaxic atlas The power spectrum of the EEG signal was analyzed using Spike2 software using a custom-written script. Monoclonal primary antibody was used MAB, Millipore, , followed by a goat anti mouse-Alexa Invitrogen, All other chemicals were from Sigma.
Microsoft Excel was used for data processing.
GraphPad Prizm was used to calculate EC50 values for concentration-response curves. How to cite this article: Tang, F. Lactate-mediated glia-neuronal signalling in the mammalian brain.
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Intervening in glial cells protects neurons in Parkinson's model | EurekAlert! Science News
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Structurally defined signaling in neuro‐glia units in the enteric nervous system
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Related Glial Neuronal Signaling
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