Journal: The Journal of Neuroscience

Article Title: Sensory Axon Growth Requires Spatiotemporal Integration of CaSR and TrkB Signaling

doi: 10.1523/JNEUROSCI.0027-19.2019

Figure Lengend Snippet: CaSR traffics together with TrkB. A, Kymographs of cotrafficking CaSR or CaSR-DN and TrkB in unstimulated and stimulated conditions from TIRF time-lapse images of MEFs cotransfected with CaSR-GFP or CaSR-DN-GFP and TrkB-RFP. B, Quantitation of comovement of CaSR and TrkB. Both, CaSR and TrkB when stimulated individually are able to increase comovement. This comovement further increases when both receptors are active simultaneously. A, C, Comovement decreases when TrkB is overexpressed with CaSR DN R185R. D, Colocalization of CaSR and TrkB under nonstimulated and stimulated conditions. Scale bar, 10 μm. Significance was determined by one-way ANOVA with post hoc Tukey. n = 25 images per condition from three independent experiments. DFn = 3, DFd = 332 (B), DFn = 3, DFd = 199 (C). Error bar indicates SEM. **p < 0.01, ***p < 0.001. Non-significant differences are indicated as n.s.

Article Snippet: On the day of imaging, MEFs were kept in “starving medium” (DMEM without penicillin/streptomycin and FBS), which was adjusted to 0.7 m m calcium for 20 min. Then, “starving media” was refreshed serving as a control, and MEFs were placed on an AxioObserver Z1 TIRF microscope (Carl Zeiss) with an Evolve CCD camera (Photometrics) using the 100× objective and imaged (5 min time-lapse recordings with pictures taken in 10 s intervals).

Techniques: Quantitation Assay