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. 2018 Jul 5;11:230. doi: 10.3389/fnmol.2018.00230

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Copyright © 2018 Wickert, Hildick, Baillie, Jelinek, Aparisi Rey, Monory, Schneider, Ross, Henley and Lutz.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Axonal polarization of HA-CB1wt and HA-CB1F238L in primary hippocampal neurons. Immunocytochemistry was performed using primary rat hippocampal neurons which were transfected with either HA-CB1wt or HA-CB1F238L. (A) Surface HA-CB1 receptor is shown in red, total HA-CB1 receptor in green and microtubule-associated protein 2 (MAP2) in blue. Surface HA-CB1 receptor was quantified in 20 × 1 μm regions of interest (ROIs) in the dendrites (MAP2+; a), proximal axon (MAP2−; b), intermediate axon (MAP2−; c) and distal axon (MAP2−; d). (B) The polarization index (A/D) for surface CB1 receptor was determined by dividing average axonal surface fluorescence by average dendritic surface fluorescence (Scale bar: 20 μm). (C) There is a significant reduction in dendritic surface CB1F238L receptor. (D) In the axon, there is a small reduction in surface CB2F238L, which did not reach statistical significance though. (Student’s t-test. Data are presented as the mean ± SEM of 21 HA-CB1wt cells and 30 HA-CB1F238L cells of two independent experiments. **p < 0.01; ***p < 0.001). (E) Intermediate and distal axonal surface CB1 receptor is expressed as percent of surface CB1 receptor in the proximal axon (repeated measures ANOVA and Bonferroni’s post hoc test. Data are presented as the mean ± SEM of 19 HA-CB1wt cells and 26 HA-CB1F238L cells of two independent experiments. *p < 0.05).