Adult Conditional Knockout of PGC-1α Leads to Loss of Dopamine Neurons

Adult conditional gene deletion of PGC-1α. A, Experimental schematic of stereotaxic intranigral virus injection and experimental time line. B, Representative GFP and TH-positive immunostaining of midbrain section from SNpc of PGC-1α^flox/flox mice injected with AAV-GFP or AAV-Cre-GFP 6 months after the injection of virus. Scale Bar, 100 µm. C, Immunoblots of PGC-1α, SDHA, Tomm20, and β-actin 4 weeks after stereotactic delivery of AAV-GFP and AAV-Cre-GFP into PGC-1α^flox/flox mice; n = 3/group. D, Quantification of C normalized to β-actin; n = 3/group. *p < 0.05, **p < 0.005. Inj, Injected.

PGC-1α isoforms. A, The illustration of PGC-1α new alternative exons and known exons in RefSeq/Ensembl/UCSC genome references. Human ESTs and mouse TSS with enhancer signals derived from >1000 human and mouse primary cells, cell lines, and tissues were mapped using CAGE. B, Diagram and PCR products show four different PGC-1α isoforms from SH-SY5Y and mouse brain and heart. C, RT-PCR for four PGC-1α isoforms, SDHA, Tomm20, and GAPDH RNA 4 weeks after stereotactic delivery of AAV-GFP or AAV-Cre-GFP into PGC-1α^flox/flox mice; n = 3/group. D, Quantification of C normalized to GAPDH; n = 3/group. *p < 0.05, **p < 0.005.

Gene deletion of PGC-1α leads to the loss of dopamine neurons in the SNpc. A, Representative TH immunohistochemistry and Nissl staining of midbrain sections from SNpc of PGC-1α^flox/flox mice injected with AAV-GFP or AAV-Cre-GFP 6 months after the injection of virus. Scale bar, 100 µm. B, Stereological assessment of TH- and Nissl-positive neurons in the SNpc of PGC-1α^flox/flox mice injected with AAV-GFP or AAV-Cre-GFP (n = 3/group). C, HPLC assessment of the striatal content of dopamine. D, HPLC assessment of the striatal content of dopamine metabolites DOPAC and HVA. E, Amphetamine-induced ipsilateral rotations (n = 3/group). Data are expressed as the mean ± SEM. *p < 0.05, **p < 0. 005. Inj, Injected; n.s, not significant.