Figure 3.
Schwann cells’ interactions with axons do not alter the patterns of bidirectional transport but increase the numbers of labeled transport vesicles.(A and D) Representative images from kymographs of vesicles transported in the anterograde (A) and retrograde (D) directions in Schwann cell/neuron cocultures are highlighted by dotted blue and red arrows, respectively. White circles highlight vesicles with just EGFP-tagged components; yellow circles show vesicles that cotransport EGFP components and NF186-mKate2. Bars: 1 µm. (B) Quantification of EGFP+ vesicles that colabel with NF186 versus the total number of anterogradely transported NF186-positive vesicles. The numbers of axons analyzed were Navβ1, n = 30; Navβ2, n = 44; and KCNQ3, n = 22. Error bars indicate mean ± SEM. (C) Quantification of anterogradely transported NF186-positive vesicles in age-matched neurons (N) cultured with or without Schwann cells (SCs) and ascorbate (vitC). The mean ± SD for each condition is N only 0.24 ± 0.43 (n = 36); N with vitC, 0.22 ± 0.51 (n = 36); N with SCs, 0.40 ± 0.60 (n = 54); and N with SCs and vitC, 1.43 ± 1.78 (n = 97). (E) Quantification of EGFP+ vesicles that colabel with NF186 versus the total number of retrogradely transported NF186-positive vesicles. The numbers of axons analyzed were Navβ1, n = 30; Navβ2, n = 44; and KCNQ3, n = 22. Error bars indicate mean ± SEM. (F) Quantification of retrogradely transported NF186-positive vesicles in age-matched neurons (N) cultured with or without Schwann cells and ascorbate. The mean ± SD for each condition is N only, 3.00 ± 1.72 (n = 46); N with vitC, 3.22 ± 2.04 (n = 27); N with SC, 3.92 ± 1.75 (n = 53); and N with SCs and vitC, 5.11 ± 2.77 (n = 96). (G) Quantification of anterogradely transported Navβ1 and Navβ2 vesicle numbers in age-matched neurons (N) cultured with or without Schwann cells and ascorbate (C). Navβ1 vesicle numbers were N only, 2.00 ± 1.41 (n = 12); N with vitC, 1.89 ± 1.54 (n = 9); N with SC, 2.10 ± 1.41 (n = 20); and N with SCs and vitC, 4.10 ± 2.29 (n = 31). Navβ2 vesicle numbers were N only, 1.90 ± 2.14 (n = 21); N with vitC, 2.00 ± 1.03 (n = 16); N with SCs, 3.33 ± 1.46 (n = 22); and N with SCs and vitC, 3.95 ± 2.17 (n = 44). (H) Quantification of retrogradely transported Navβ1 and Navβ2 vesicle numbers in age-matched neurons (N) cultured with or without Schwann cells and ascorbate (C). Navβ1 vesicle numbers were N only, 4.58 ± 1.56 (n = 12); N with vitC, 3.56 ± 1.94 (n = 9); N with SCs, 4.85 ± 1.53 (n = 20); and N with SCs and vitC, 5.40 ± 2.42 (n = 30). Navβ1 vesicle numbers were N only, 4.14 ± 2.29 (n = 21); N with vitC, 5.38 ± 2.06 (n = 16); N with SCs, 7.52 ± 2.04 (n = 21); and N with SCs and vitC, 7.48 ± 3.13 (n = 44). In panels C and F–H, statistical significance was determined by two-tailed Student’s t test for unpaired data, with asterisks indicating P values of ****, P < 0.001; ***, P < 0.005; **, P < 0.01; and *, P < 0.05; ns, not significant.

Schwann cells’ interactions with axons do not alter the patterns of bidirectional transport but increase the numbers of labeled transport vesicles. (A and D) Representative images from kymographs of vesicles transported in the anterograde (A) and retrograde (D) directions in Schwann cell/neuron cocultures are highlighted by dotted blue and red arrows, respectively. White circles highlight vesicles with just EGFP-tagged components; yellow circles show vesicles that cotransport EGFP components and NF186-mKate2. Bars: 1 µm. (B) Quantification of EGFP+ vesicles that colabel with NF186 versus the total number of anterogradely transported NF186-positive vesicles. The numbers of axons analyzed were Navβ1, n = 30; Navβ2, n = 44; and KCNQ3, n = 22. Error bars indicate mean ± SEM. (C) Quantification of anterogradely transported NF186-positive vesicles in age-matched neurons (N) cultured with or without Schwann cells (SCs) and ascorbate (vitC). The mean ± SD for each condition is N only 0.24 ± 0.43 (n = 36); N with vitC, 0.22 ± 0.51 (n = 36); N with SCs, 0.40 ± 0.60 (n = 54); and N with SCs and vitC, 1.43 ± 1.78 (n = 97). (E) Quantification of EGFP+ vesicles that colabel with NF186 versus the total number of retrogradely transported NF186-positive vesicles. The numbers of axons analyzed were Navβ1, n = 30; Navβ2, n = 44; and KCNQ3, n = 22. Error bars indicate mean ± SEM. (F) Quantification of retrogradely transported NF186-positive vesicles in age-matched neurons (N) cultured with or without Schwann cells and ascorbate. The mean ± SD for each condition is N only, 3.00 ± 1.72 (n = 46); N with vitC, 3.22 ± 2.04 (n = 27); N with SC, 3.92 ± 1.75 (n = 53); and N with SCs and vitC, 5.11 ± 2.77 (n = 96). (G) Quantification of anterogradely transported Navβ1 and Navβ2 vesicle numbers in age-matched neurons (N) cultured with or without Schwann cells and ascorbate (C). Navβ1 vesicle numbers were N only, 2.00 ± 1.41 (n = 12); N with vitC, 1.89 ± 1.54 (n = 9); N with SC, 2.10 ± 1.41 (n = 20); and N with SCs and vitC, 4.10 ± 2.29 (n = 31). Navβ2 vesicle numbers were N only, 1.90 ± 2.14 (n = 21); N with vitC, 2.00 ± 1.03 (n = 16); N with SCs, 3.33 ± 1.46 (n = 22); and N with SCs and vitC, 3.95 ± 2.17 (n = 44). (H) Quantification of retrogradely transported Navβ1 and Navβ2 vesicle numbers in age-matched neurons (N) cultured with or without Schwann cells and ascorbate (C). Navβ1 vesicle numbers were N only, 4.58 ± 1.56 (n = 12); N with vitC, 3.56 ± 1.94 (n = 9); N with SCs, 4.85 ± 1.53 (n = 20); and N with SCs and vitC, 5.40 ± 2.42 (n = 30). Navβ1 vesicle numbers were N only, 4.14 ± 2.29 (n = 21); N with vitC, 5.38 ± 2.06 (n = 16); N with SCs, 7.52 ± 2.04 (n = 21); and N with SCs and vitC, 7.48 ± 3.13 (n = 44). In panels C and F–H, statistical significance was determined by two-tailed Student’s t test for unpaired data, with asterisks indicating P values of ****, P < 0.001; ***, P < 0.005; **, P < 0.01; and *, P < 0.05; ns, not significant.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal