Transporter proteins as percent of total tissue protein
Hippocampus | Cerebellum | ||||
---|---|---|---|---|---|
Whole | CA1 Stratum radiatum | Whole | Whole | Stratum moleculare | |
GLAST | A | C | A | B | D |
1 | 0.25 ± 0.02 | 0.40 ± 0.01 | 0.81 ± 0.03 | 1.0 ± 0.04 | 2.3 ± 0.05 |
2 | 0.13 ± 0.01 | 0.21 ± 0.004 | 0.47 ± 0.04 | 0.53 ± 0.02 | 1.3 ± 0.03 |
3 | 0.24 ± 0.01 | 0.36 ± 0.01 | 0.70 ± 0.05 | 0.79 ± 0.03 | 2.0 ± 0.04 |
4 | 0.64 ± 0.03 | 1.6 ± 0.03 | |||
Mean | 0.21 ± 0.02 | 0.32 ± 0.01 | 0.66 ± 0.04 | 0.74 ± 0.03 | 1.8 ± 0.02 |
GLT | B | C | B | D | |
1 | 1.3 ± 0.03 | 1.3 ± 0.03 | 0.27 ± 0.005 | 0.34 ± 0.004 | |
2 | 1.2 ± 0.02 | 1.3 ± 0.04 | 0.24 ± 0.005 | 0.30 ± 0.01 | |
3 | 1.1 ± 0.03 | 1.3 ± 0.04 | 0.31 ± 0.01 | ||
4 SDS | 1.2 ± 0.02 | 1.2 ± 0.03 | 0.22 ± 0.004 | 0.28 ± 0.003 | |
5 | 1.1 ± 0.02 | 1.1 ± 0.03 | 0.22 ± 0.003 | 0.27 ± 0.001 | |
Mean | 1.2 ± 0.02 | 1.3 ± 0.03 | 0.24 ± 0.004 | 0.30 ± 0.01 |
The contents of the glutamate transporters in tissue samples (whole hippocampus, stratum radiatum of CA1 hippocampus, whole cerebellum, and stratum moleculare of the cerebellum) were determined by quantitative immunoblotting as described in Materials and Methods using purified transporter protein as standard. Each series of SDS extracts (A–D) contained tissue from three rats. The immunoreactivities of these extracts (which contained all tissue proteins) were compared with that of different batches of purified transporter protein (GLAST: four batches; GLT: five batches). The results shown above are given as percent (mean ± SEM of the three rats; n = 3) of total tissue protein. The GLT protein was isolated from the entire forebrain solubilized with cholate (or SDS as indicated).