Table 1.
Prior western blot studies of mGlu3 in schizophrenia, showing main methodological features and key findings.
| Study | Antibody | Antibody validation | Sample | Sample preparation and methoda | Brain areab | Loading controls | Bands measured | Main findings |
|---|---|---|---|---|---|---|---|---|
| Crook et al. (2002) | Anti-mGlu2/3 (Chemicon) | Pre-absorption with mGlu2/3 peptide abolished immunoreactivity. | 20 SCZ, 20 CON | Membranes; 7.5% PA gels and 50 μg protein (duplicates). | BA46 | Two internal controls in each blot. | ~ 100 kDa | No group difference. Negative correlation with age in controls. |
| Gupta et al. (2005) | Anti-mGlu2/3 (Upstate Bio-technology) | No data presented; paper refers to earlier papers, but the latter used a Chemicon mGlu2/3 antibody (with peptide pre-absorption used as the control). | 16 SCZ, 9 CON | Total homogenates; 5% β-ME and heated at 95 °C for 4 min; 7.5% PA gels and 40 μg protein (duplicates). | BA9, 11, 32 and 46; n. accumbens, putamen, caudate | None stated. | ~ 100–110 kDa | Increase in BA46 in schizophrenia. |
| Corti et al. (2007) | Anti-mGlu3, raised against residues 16–35 in mouse mGlu3. | Tested using GRM3 cDNA-transfected CHO cells, and grm3−/− mouse brain. | 20 SCZ, 35 CON | Membranes; 20 mM DTT, heated at 60 °C for 3 min; 8% PA gels 40 μg protein (triplicates). | BA10 | β-actin and Ponceau S. Each gel also contained two internal standards. | ~ 100 and ~ 200 kDa bands in cells and WT mouse brain; not seen in grm3−/− mouse. Human brain: bands at ~ 200 kDa and doublet at ~ 95/100 kDa. | 200 kDa band reduced in schizophrenia. Negative correlations with age for 200 kDa and 100 kDa bands. |
| Ghose et al. (2009) | Anti-mGlu3 (Abcam) | No data presented; the authors refer to a prior paper, but the latter does not provide clarity. | 15 SCZ, 15 CON | Total homogenates; 10% PA gels and 20 μg protein (duplicates). | Prefrontal, temporal and motor cortex. | β-tubulin | Monomer; Molecular weight not specified. | mGlu3 decreased in prefrontal cortex in schizophrenia. |
a
DTT: dithiothreitol. ME: mercaptoethanol. PA: polyacrylamide.
b
BA: Brodmann area. BA9/46: dorsolateral prefrontal cortex. BA10: frontal pole. BA11, BA32: medial prefrontal cortex.