Fig. 7. PANTHOS neurodegeneration coincides with β-amyloid plaque formation and subsequent lysosomal neuronal cell death.
a, Aβ antibody 3D6 detecting the appearance of amyloid plaques in 5xFAD mice (2.7-month-old male) demonstrates co-incidence with the presence of a PANTHOS neuron. Scale bar, 20 μm. b, Quantitative percentage of PANTHOS neurons that are 3D6-positive (top) and percentage of PANTHOS among 3D6-positive plaques that are associated with PANTHOS (bottom)—with PANTHOS (91.7 ± 0.5%), without PANTHOS (8.3 ± 0.5%), with 3D6 (100 ± 0%), without 3D6 (0 ± 0%). n = 3 mice. c, DAPI staining depicting various stages of PANTHOS development and ultimate disappearance of detectable DAPI (although not necessarily nuclear marker IR; see Extended Data Fig. 5e). Normal DAPI-labeled nucleus (double arrow), condensed DAPI signal (single arrow) and non-detectable DAPI in very advanced PANTHOS neuron (arrowhead). Scale bar, 10 μm. d, Percentage of PANTHOS neurons with detectable DAPI label in 2.7-month-old or 6-month-old 5xFAD/TRGL mouse brain. 2.7 months: with DAPI (91.4 ± 1.3%) and without DAPI (8.6 ± 1.3%); 6 months: with DAPI (67.8 ± 4.5%) and without DAPI (32.2 ± 4.5%). n = 6 (two sections per mouse, three mice; 94 neurons in cortex area were counted). e, Lysosomal enzyme distribution in cytosol and membrane/vesicle fraction in 2.7-month-old and 6-month-old 5xFAD and WT male mouse cortex. Cytosolic CTSD: 2.7 months (99.8 ± 1.9%) and 6 months (260.4 ± 3.1%); cytosolic CTSB: 2.7 months (103.8 ± 1.6%) and 6 months (238.5 ± 5.9%). n = 3 mice per each genotype. f, Immunofluorescence labeling of 2.7-month-old 5xFAD/TRGL mouse brain neurons with a CTSD antibody. Arrow indicates normal CTSD-positive puncta in a healthy neuron. The experiment was repeated three times independently with similar results. The arrowhead indicates diffuse CTSD signal in a PANTHOS neuron. Scale bar, 20 μm. Quantitative data are presented as means ± s.e.m., unpaired t-test, two-tailed P value as indicated. mo, month; rel., relative.