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. 2021 Jan 6;10:e63092. doi: 10.7554/eLife.63092

Figure 6. Local protein synthesis occurs in the dendrites of GABAergic neurons.

Figure 6.

(A) Schematic outline of the experiment addressing translational capacity of GABAergic neurites (in GAD-Cre::GFP-2A-MetRS*, Sst-Cre::GFP-2A-MetRS*) and glutamatergic neurites (Nex-Cre::GFP-2A-MetRS*) with cell type-specific ANL incorporation and BONCAT for results shown in B–E. (B) Representative biotin immunoblot for an experiment with Sst-Cre::GFP-2A-MetRS* mouse cortical neurons as outlined in A. (C–E) Quantification of the relative biotin western blot signal representing 15 min ANL incorporation in the soma (left bars) or neurite (right bars) fractions harvested under the conditions shown in (A) from cortical neuron cultures prepared from the indicated mouse lines: (C) GAD-Cre::GFP-2A-MetRS*, (D) Sst-Cre::GFP-2A-MetRS*, and (E) Nex-Cre::GFP-2A-MetRS* (mean ± SD; GAD: soma no 0.61 ±0.07, soma neu 0.65 ± 0.06, soma full 1.00 ± 0.07, neurites no 0.68 ± 0.06, neurites neu 0.96 ± 0.07, neurites full 1.00 ± 0.19; Sst: soma no 0.47 ± 0.17, soma neu 0.47 ± 0.1, soma full 1.00 ± 0.15, neurites no 0.43 ± 0.13, neurites neu 0.93 ± 0.2, neurites full 0.73 ± 0.19; Nex: soma no 0.71 ± 0.11, soma neu 0.68 ± 0.10, soma full 1.00 ± 0.12, neurites no 0.65 ± 0.09, neurites neu 0.89 ± 0.13, neurites full 0.91 ± 0.12). Equal protein amounts were loaded within soma fractions and within neurite fractions. The biotin immunoreactivity in the neurite fraction of the neurite-only labeling from all investigated mouse lines was significantly higher than in unlabeled controls ([Kruskal–Wallis test with Dunn’s multiple comparison’s test for GAD, n = 6 (three experiments); GAD soma: soma no vs soma neu p>0.9999, soma no vs soma full p=0.0032, soma neu vs soma full p=0.0304; GAD neurites: neurites no vs neurites neu p=0.0239, neurites no vs neurites full p=0.0073, neurites neu vs neurites full p>0.9999; Sst n = 7 (three experiments); Sst soma: soma no vs soma neu p>0.9999, soma no vs soma full p=0.0023, soma neu vs soma full p=0.0089; Sst neurites: neurites no vs neurites neu p=0.0008, neurites no vs neurites full p=0.0635, neurites neu vs neurites full p=0.5449]; Nex n = 8 [four experiments] Nex soma: soma no vs soma neu p>0.9999, soma no vs soma full p=0.0036, soma neu vs soma full p=0.0017; Nex neurites: neurites no vs neurites neu p=0.0050, neurites no vs neurites full p=0.0080, neurites neu vs neurites full p>0.9999).