Conditional deletion of the glutamate transporter GLT-1 reveals that astrocytic GLT-1 protects against fatal epilepsy while neuronal GLT-1 contributes significantly to glutamate uptake into synaptosomes
| Autor: | Paul A. Rosenberg, Kathryn D. Fischer, Niels C. Danbolt, Clive Miranda, Wencke Armsen, Geraldine T. Petr, Yun Zhou, Natalie M. Frederick, Mustafa Q. Hameed, Edward A. Bedoya, Sameer C. Dhamne, Yan Sun, Alexander Rotenberg, Jianlin Wang, Chiye Aoki |
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| Rok vydání: | 2015 |
| Předmět: |
Male
medicine.medical_specialty Ischemia Presynaptic Terminals Glutamic Acid Biology Epilepsy Mice Internal medicine medicine Animals Gene Mice Knockout Neurons Liposome General Neuroscience Body Weight SLC1A2 Brain Electroencephalography Glutamic acid Articles medicine.disease Endocrinology Biochemistry Excitatory Amino Acid Transporter 2 Astrocytes Forebrain Liposomes Excitatory postsynaptic potential biology.protein Female Synaptosomes |
| Zdroj: | The Journal of neuroscience : the official journal of the Society for Neuroscience. 35(13) |
| ISSN: | 1529-2401 |
| Popis: | GLT-1 (EAAT2; slc1a2) is the major glutamate transporter in the brain, and is predominantly expressed in astrocytes, but at lower levels also in excitatory terminals. We generated a conditional GLT-1 knock-out mouse to uncover cell-type-specific functional roles of GLT-1. Inactivation of the GLT-1 gene was achieved in either neurons or astrocytes by expression of synapsin-Cre or inducible human GFAP-CreERT2. Elimination of GLT-1 from astrocytes resulted in loss of ∼80% of GLT-1 protein and of glutamate uptake activity that could be solubilized and reconstituted in liposomes. This loss was accompanied by excess mortality, lower body weight, and seizures suggesting that astrocytic GLT-1 is of major importance. However, there was only a small (15%) reduction that did not reach significance of glutamate uptake into crude forebrain synaptosomes. In contrast, when GLT-1 was deleted in neurons, both the GLT-1 protein and glutamate uptake activity that could be solubilized and reconstituted in liposomes were virtually unaffected. These mice showed normal survival, weight gain, and no seizures. However, the synaptosomal glutamate uptake capacity (Vmax) was reduced significantly (40%). In conclusion, astrocytic GLT-1 performs critical functions required for normal weight gain, resistance to epilepsy, and survival. However, the contribution of astrocytic GLT-1 to glutamate uptake into synaptosomes is less than expected, and the contribution of neuronal GLT-1 to synaptosomal glutamate uptake is greater than expected based on their relative protein expression. These results have important implications for the interpretation of the many previous studies assessing glutamate uptake capacity by measuring synaptosomal uptake. |
| Databáze: | OpenAIRE |
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