Figure 7. Long-Term Progerin Overexpression In Vivo Reveals a Severe Degenerative Phenotype in PD Mutant Cells.

(A) Schematic illustration of the transplantation studies into 6-OHDA lesioned Parkinsonian mice.

(B) Rotational behavior analysis of lesioned mice transplanted with control or PD mutant iPSC-mDA neurons expressing hSyn::nuclear-GFP or hSyn::GFP-progerin. Mice were lesioned and tested for amphetamine-induced rotation behavior twice prior to grafting. Dotted line indicates threshold for successful lesioning. Pink symbols identify successfully lesioned animals that did not show recovery. n= 3-5 animals per treatment group.

(C) Assessment at 3 months post transplant revealed a dramatic loss of TH+ mDA neurons in PD mutants overexpressing progerin.

(D) Quantification of the percentage of GFP+ cells that are TH+. Data are presented as mean ± SEM. n= 3 mice per condition.

(E-G) Ultrastructural analysis 6 months after transplantation revealed accumulation of neuromelanin with lipofuscin deposits (E, yellow arrowheads) in grafts with progerin overexpression. Strikingly, the PINK1 mutant graft with progerin displayed enlarged mitochondria (F, compare representative mitochondria indicated by orange arrows in +nuclear-GFP and +GFP-progerin groups) while the Parkin mutant graft with progerin had large multilamellar bodies (G, pink arrows). These phenotypes were not observed in any other treatment groups. Asterisks in (E) and (G) indicate a fibrillar body.

*p<0.05, ** p<0.01 according to Student’s t-tests. Bar graph represents mean ± SEM. Scale bars: 200 μm (C), 500 nm (E-G).

See also Figure S6-S7 & Tables S2 and S4-S5.