Fig. 4

Morphological analysis of α-syn-positive inclusions upon macroautophagy blocking treatment in oligodendroglial cells. Confocal microscopy was performed to analyze the incorporation and inclusion formation of sol and fib α-syn in oligodendroglial cells challenged with BAF for 24 h. Immunocytochemistry was accomplished following fixation, labelling human α-syn (15G7, red), the membrane using wheat germ agglutinin (WGA, green) and the nucleus using DAPI (blue). Treatment with BAF induced a more pronounced vesicle formation as can be seen by the membrane staining with WGA (a). The addition of recombinant sol and fib α-syn for 24 h to the medium induced the incorporation of α-syn by oligodendroglial cells. Yet, simultaneous macroautophagy blocking did not induce an increased amount of α-syn incorporated by these oligodendroglial cells. No GCI-like formation was observed upon macroautophagy block and treatment with extracellularly added sol or fib α-syn. In some cells translocation of α-syn to the nucleus was observed as shown in picture fib α-syn. Moreover, BAF treatment induced an enhanced number of vesicles in the cytoplasm (WGA staining) irrespective of α-syn in the cytoplasm of the cells (b). Scale bar 10 µm