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. 2017 Dec 6;37(49):11835–11853. doi: 10.1523/JNEUROSCI.0983-17.2017

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Copyright © 2017 Rostami et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License Creative Commons Attribution 4.0 International, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Figure 8. — Healthy astrocytes deliver mitochondria to α-SYN oligomer-exposed astrocytes. Using Mitotracker transfection, we could demonstrate that mitochondria were present within TNTs connecting the astrocytes (A). A close-up of the white rectangle is shown in A′. Coculture experiments were performed using two different set-ups. Either, GFAP::tRFP astrocytes (red acceptor cells) were cocultured with Mitotracker-transfected, α-SYN oligomer-exposed astrocytes (donor cells) (B) or α-SYN oligomer-exposed astrocytes (acceptor cells) were cocultured with GFAP::tRFP, Mitotracker-transfected astrocytes (red donor cells) (C). Using immunocytochemistry, we identified transfer of mitochondria between the GFAP::tRFP astrocytes and the unlabeled astrocytes via direct contact (dashed areas show the outline of separate cells) (D) and TNTs (E), a close-up of the white rectangle is shown in E′. Quantification of the number of Mitotracker⁺ acceptor cells in the two coculture setups indicated that the healthy astrocytes transferred significantly more mitochondria to α-SYN oligomer-exposed cells than the other way around (F). Addition of latrunculin B to the cocultures significantly decreased the percentage of Mitotracker⁺ acceptor cells (G). In addition, basal level of mitochondrial transfer was investigated in untreated control cultures. Compared with α-SYN oligomer-exposed cultures, the number of Mitotracker⁺ acceptor cells was significantly lower in the control cultures (see Fig. 8-1). Scale bars: A, 20 μm; D, E, 20 μm. Data are presented as mean ± SEM from three independent experiments and the levels of significance were set to *p < 0.05, **p < 0.01, and ***p < 0.001 (F, G).