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. 2023 Nov 13;16:1241420. doi: 10.3389/fnmol.2023.1241420

Figure 5.

Figure 5

Retrograde transport of proNGF-KKE is impaired with in vitro age in BFCNs. 50pM of quantum dot labeled proNGF-KKE was added to the axon terminals of young (DIV8-9) or aged (DIV20-22) rat BFCNs for 1 h prior to analysis of quantum dot accumulation at BFCN axon terminals, proximal axons, and cell bodies. proNGF-KKE is shown in yellow, tubulin is shown in red, and DAPI is shown in blue. (A) ProNGF-KKE was observed in young but not aged BFCN cell bodies. White arrows indicate quantum dots that accumulated in the proximal axons and cell bodies. Aged BFCNs exhibited decreased accumulation of proNGF-KKE at the proximal axons and increased accumulation of proNGF-KKE at the axon terminals compared to young BFCNs. (B,C) Quantification of the images shown in A. (B) Proximal axons: young n = 30 images, aged n = 28 images. The highest two points in the DIV20-22 group were detected as outliers using ROUT (Q = 1%). Removal of these outliers did not change the observed value of p. (C) Axon terminals: young n = 30 images, aged n = 37 images. No outliers were detected using ROUT (Q = 1%). All sample sizes are taken from three separate chambers in three independent experiments. Error bars represent SEM. ****p < 0.0001. DIV, days in vitro; QD, quantum dot. Scale bars: 10 μm.