Fig. 6

Mitochondrial dysfunction and actin cytoskeletal pathology in mammalian LRRK2-G2019S models. a, Morphology and expression of neuronal ß-tubulin (TuJ1) in cortical neurons derived from LRRK2-G2019S heterozygous iPSC and isogenic controls. Scale bars represent 40 µm in (a). b-d, LRRK2-G2019S heterozygous human neurons have increased F-actin as assessed by staining with fluorescent phalloidin (b), as well as numerous actin aggregates (arrows) as quantified in (c,d). Scale bar in (b) represents 15 µm. n=6 in (c), n=3 in (d). e-h, Metabolic profiling of cortical neurons derived from LRRK2-G2019S heterozygous iPSC and isogenic controls in Seahorse XF 96-well culture microplates reveals decreased oxygen consumption rate (e, OCR), basal respiration (f), the energy phenotype as determined by simultaneously the OCR and extracellular acidification rate (ECAR) from the same experiment, and non-mitochondrial respiration (h). n=4 in (f-h). i, Cortical brain sections from LRRK2-G2019S heterozygous knockin mice have increased F-actin as assessed by staining with fluorescent phalloidin and quantified in (j). Scale bar in (i) represents 75 µm. k, Native gel electrophoresis followed by western blotting for LRRK2 reveals an increased multimer to monomer ratio in cortical homogenates from LRRK2-G2019S heterozygous knockin mice, as quantified in (l). n=6 per genotype in (i-l). Data are represented as mean ± SD. *p<0.05, **p<0.01, ***p<0.005, Student’s t-test. Mice are 10-12 weeks old