Figure 1

Real-Time PCR amplification of Glutamate receptor (mGlu-5) mRNA from the Cerebral cortex of Control, Diabetic and Insulin treated Diabetic Rats. Sample Names. 1. 7 weeks Control, 2. 7 weeks Diabetic, 3. 7 weeks Insulin treated Diabetic. 4. 90 weeks Control, 5. 90 weeks Diabetic, 6. 90 weeks Insulin treated Diabetic. Real Time PCR analysis was done using mGlu-5 specific primer and fluorescently labeled Taq probe. The TaqMan reaction mixture of 20 μl contained 25 ng of total RNA-derived cDNAs, 200 nM each of the forward primer, reverse primer and TaqMan probe for glutamergic- (mGlu-5) gene, endogenous control, β-actin and 12.5 μl of TaqMan 2× Universal PCR Mastermix (Applied Biosystems). All reactions were performed in duplicates. The ΔΔCT method of relative quantification was used to determine the fold change in expression. This was done by first normalizing the resulting threshold cycle (CT) values of the target mRNAs to the CT values of the internal control β-actin in the same samples (ΔCT = CT_Target - CT_β-actin). It was further normalized with the control (ΔΔCT = ΔCT - CT_Control). The fold change in expression was then obtained as (2^-ΔΔCT) and expressed as log 2^-ΔΔCT. Values are Mean ± S.D of 4-6 separate experiments. ***p < 0.001 when compared to control, ^¶¶¶ p < 0.001 when compared to diabetic, ^††† p < 0.001 when compared to 7 weeks old diabetic, ^aaa p < 0.001 when compared to 7 weeks old insulin treated diabetic.