Figure 2.
Figure 2. Active lysosomes are found in distal dendrites. (A) Lysosome membrane disruption by GPN abolishes LysoTracker staining. Representative dissociated hippocampal neurons expressing GFP (Sindbis virus) and costained with LysoTracker. Images were taken before adding either vehicle (DMSO) or 40 µM GPN. Images were taken 5 min after either vehicle or GPN. Regions of interest (boxes) are magnified below. (B) Representative neuron transfected with mCherry and GCaMP3-TRPML1. Neuron was imaged live for 100 s in the 488 and 568 channel. (C) Straightened segment from B showing GCaMP3-TRPML1 signal 2 s after adding either vehicle or 40 µM GPN. (D) Detection of calcium release from lysosome stores by GCaMP3-TRPML1 in representative neuron transfected with mCherry and GCaMP3-TRPML1 from B and C. Vehicle was added 50 s after the start of the recording, and 40 µM GPN was added 80 s after the start of the recording. Asterisks show time where images from D were taken.

Active lysosomes are found in distal dendrites. (A) Lysosome membrane disruption by GPN abolishes LysoTracker staining. Representative dissociated hippocampal neurons expressing GFP (Sindbis virus) and costained with LysoTracker. Images were taken before adding either vehicle (DMSO) or 40 µM GPN. Images were taken 5 min after either vehicle or GPN. Regions of interest (boxes) are magnified below. (B) Representative neuron transfected with mCherry and GCaMP3-TRPML1. Neuron was imaged live for 100 s in the 488 and 568 channel. (C) Straightened segment from B showing GCaMP3-TRPML1 signal 2 s after adding either vehicle or 40 µM GPN. (D) Detection of calcium release from lysosome stores by GCaMP3-TRPML1 in representative neuron transfected with mCherry and GCaMP3-TRPML1 from B and C. Vehicle was added 50 s after the start of the recording, and 40 µM GPN was added 80 s after the start of the recording. Asterisks show time where images from D were taken.

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