Fig. 5
From: Granulovacuolar degeneration bodies are independently induced by tau and α-synuclein pathology
GVBs are found in association with pathological α-synuclein accumulation in the seeded model. a Schematic representation of the protocol to induce intraneuronal α-syn pathology in primary mouse neurons. WT neurons were plated at DIV 0. At DIV 7, α-syn PFFs were added to the culture medium. Cells were fixed and analysed by immunofluorescent staining for pathological α-syn and GVBs at DIV 18. b Representative confocal images showing an α-syn+/GVB+ (left) and an α-syn−/GVB− control neuron (right) to which the seed vehicle (PBS) was added. Immunofluorescence staining was performed for the neuron-specific dendritic marker MAP2 (grey), phosphorylated α-syn (p-αsyn129, green) and GVB marker CK1δ (magenta). Separate channels are shown in greyscale. c, d Representative confocal images of GVB+ neurons in the α-syn model. Co-immunofluorescence staining was performed for the canonical GVB marker CK1δ (magenta) and the additional GVB core marker pPERK (c) or the GVB membrane marker LIMP2 (d) (green). Nuclei are visualised by DAPI (blue). Separate channels are shown in greyscale and in colour in the merge. A line intensity profile along the white bar indicated in the merge shows CK1δ signal overlap with pPERK and being surrounded by LIMP2