The Journal of Neuroscience, January 11, 2006, ():
Impaired Glutamate Transport in a Mouse Model of Tau Pathology in Astrocytes
J. Neurosci. Dabir et al.
26: 644
Supplemental data
Files in this Data Supplement:
- supplemental material
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Supplemental Figure S1. GLT-1 and GLAST immunoreactivity in cortices of GFAP/tau Tg mice.
Immunohistochemical analysis was performed on cortical sections from 24-month-old non-Tg, GFAP/tauWT (WT) and GFAP/tauP301L (PL) mice, as indicated, with antibodies to GLT-1 and GLAST. Patchy staining for GLT-1 and GLAST were detected in both strains of tau Tg mice compared to the diffuse gray matter staining observed in non-Tg animals. However, there was no significant change in GLT-1 or GLAST protein expression detected in the cortex of both lines of Tg mice (see Fig. 4B). Scale bar = 100 μm.
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Supplemental figure S2. Decreased glial glutamate transporter expression in GFAP/tau Tg mice.
Representative immunoblot analysis of glutamate transporter protein expression in cortex, brainstem and spinal cords of 24 month old GFAP/tauWT, GFAP/tauP301L and non-Tg mice. Twenty five (T14, GLT-1, and actin) to 40 μg (GLAST and EAAC1) of total protein extract were resolved by SDS-PAGE and detected with antibodies to human tau (T14), GLT-1, GLAST, EAAC1 and actin. Arrowheads and asterisks indicate the monomeric (arrowheads), dimeric (asterisk, EAAC1) or multimeric (asterisk, GLT-1) forms of the transporter protein. The immunoblots were quantified using Multigauge v2.3 software and the levels of each of the transporters were normalized to actin levels. At 24 months of age, there is a decrease in both GLT-1 and GLAST in the brainstem and spinal cord of GFAP/tauWT and GFAP/tauP301L Tg mice relative to non-Tg animals (see Fig. 4 for quantification). There is no significant difference in neocortex where only limited tau pathology is detected. Furthermore, there is no change in relative protein expression of the neuronal glutamate transporter EAAC1.
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Supplemental figure S3. Decreased GLT-1 expression in affected cortices of CBD patients.
Frontal cortex gray matter from CBD, AD, and normal brains was extracted as described in the Material and Methods section. Fifty μg of total protein extract was resolved by SDS-PAGE and detected with antibodies to GLT-1 and actin. Arrowheads and asterisks indicate the monomeric and multimeric forms of GLT-1, respectively. The immunoblots were quantified using Multigauge v2.3 software and the levels of GLT-1 were normalized to actin levels. In all three CBD cases, there is decrease in GLT-1 protein expression relative to the normal brain, and in two of these cases the reduced expression is approximately 50%. The level of GLT-1 protein expression was not altered in AD brain tissue relative to controls.
