Abstract
Granulovacuolar degeneration bodies (GVBs) are membrane-bound vacuolar structures harboring a dense core that accumulate in the brains of patients with neurodegenerative disorders, including Alzheimer's disease and other tauopathies. Insight into the origin of GVBs and their connection to tau pathology has been limited by the lack of suitable experimental models for GVB formation. Here, we used confocal, automated, super-resolution and electron microscopy to demonstrate that the seeding of tau pathology triggers the formation of GVBs in different mouse models in vivo and in primary mouse neurons in vitro. Seeding-induced intracellular tau aggregation, but not seed exposure alone, causes GVB formation in cultured neurons, but not in astrocytes. The extent of tau pathology strongly correlates with the GVB load. Tau-induced GVBs are immunoreactive for the established GVB markers CK1δ, CK1ɛ, CHMP2B, pPERK, peIF2α and pIRE1α and contain a LAMP1- and LIMP2-positive single membrane that surrounds the dense core and vacuole. The proteolysis reporter DQ-BSA is detected in the majority of GVBs, demonstrating that GVBs contain degraded endocytic cargo. GFP-tagged CK1δ accumulates in the GVB core, whereas GFP-tagged tau or GFP alone does not, indicating selective targeting of cytosolic proteins to GVBs. Taken together, we established the first in vitro model for GVB formation by seeding tau pathology in primary neurons. The tau-induced GVBs have the marker signature and morphological characteristics of GVBs in the human brain. We show that GVBs are lysosomal structures distinguished by the accumulation of a characteristic subset of proteins in a dense core.
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📊 Figures
Fig.u00a01
Seeding of tau pathology induces GVBs in vivo. a u2013 g Immunofluorescence was performed on hippocampal sections of tau P301L Tg mice injected with K18 tau P301L seeds (PFF, N =u20095) or control buf...
Fig.u00a02
Seeding of tau pathology induces GVBs in cultured primary neurons. a Representative confocal images and zooms of primary mouse hippocampal neurons expressing tau-P301L-GFP (green) treated with K18 tau...
Fig.u00a03
Intracellular tau pathology causes GVB formation. a u2013 c Primary mouse neurons with (+) or without (u2212) expression of tau-P301L in the presence (PFF) or absence (Ctrl) of seeds. Representative e...
Fig.u00a04
GVBs are not detected in cultured primary astrocytes with seeding-induced tau pathology. a , b Primary mouse neurons ( a ) and astrocytes ( b ) expressing tau-P301L-mCherry were cultured separately an...
Fig.u00a05
Tau-induced GVBs are positive for late autophagic and endocytic markers. Tau pathology was induced in primary mouse neurons expressing tau P301L by treatment with PFFs. a u2013 c Representative confoc...
Fig.u00a06
GVBs are lysosomal structures that contain degraded endocytic cargo. Tau pathology was induced in primary mouse neurons expressing tau P301L by treatment with PFFs. a Representative confocal images pl...
Fig.u00a07
Tau-induced GVBs contain one or more cores enclosed by a single LIMP2-positive membrane. Tau pathology was induced in primary mouse neurons expressing tau P301L by treatment with PFFs and GVBs were vi...
Fig.u00a08
Specific cytosolic cargo accumulates in the core of GVBs. Tau pathology was induced in primary mouse neurons expressing tau P301L by treatment with PFFs. a , b Representative confocal images and fluor...
Fig.u00a09
GVBs are lysosomal structures that contain accumulated endocytic and autophagic cargo. Schematic representation of the autophagic and endocytic pathways and in the inset the summary of the characteriz...
Figure images are served from the NIH/NLM PubMed Central Open Access Subset or Europe PMC; copyright remains with the publishers and authors.
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