Figure 1.
Development of neurons with axon-carrying dendrite. (A) Representative images of dissociated hippocampal primary neurons with nonAcD (top row) and AcD (bottom row) morphology at different ages. AnkG and MAP2 immunostaining indicate the axon/AIS and dendrites of a neuron, respectively. White arrowhead indicates AIS; yellow and pink arrowhead indicates AcD and stem dendrite of AcD neuron respectively; and cyan arrowhead indicates regular dendrite. Scale bar is 20 µm. (B) Schematic of AcD neuron. Black dashed line indicates the end of soma; orange dashed line indicates the beginning of an axon. Cyan solid line with double arrowheads indicates the axon distance. Magenta solid line with double arrowheads indicates the first (d1) and the second (d2) line used to calculate the averaged diameter of stem dendrite. (C) Time-lapse images of nonAcD (top row) and AcD neurons (bottom row) at different developmental stages. The precursor neurite in the displayed AcD neuron became an axon and collateral from the precursor neurite developed as an AcD. Orange arrowhead indicates the precursor neurite of AcD neuron; blue arrowhead indicates the precursor neurite of nonAcD neuron; white arrowhead indicates axon; yellow and pink arrowhead indicates AcD and stem dendrite of AcD neuron, respectively. Scale bar is 20 µm. Cell bodies with dendrites and the AIS are labeled by MAP2 and AnkG staining, respectively. Related to Videos 1 and 2. (D) Percentage of AcD neurons with a generated collateral or bifurcation of the precursor neurite. 1 culture preparation, AcD (total): n = 27 cells, collateral: n = 22 cells (∼80%), bifurcation: n = 5 cells (∼20%). (E) Time points of collateral formation at the precursor neurite in AcD neurons. One neuronal culture preparation, AcD (total): n = 27 cells, time points: 27 h (n = 3 cells), 33 h (n = 7 cells), 39 h (n = 9 cells), 45 h (n = 4 cells), 51 h (n = 4 cells). (F) Percentage of AcD neurons developing the precursor neurite into an axon or an AcD. 1 culture preparation, AcD: n = 22 cells, precursor as axon: n = 16 cells (∼70%), precursor as AcD: n = 6 cells (∼30%). (G) Percentage of nonAcD and AcD neurons that formed collaterals at the proximal region of the precursor neurite during development. One culture preparation, nonAcD: n = 25 cells (percentage of neurons with collaterals versus without collaterals: ∼32% versus ∼68%), AcD: n = 23 cells (percentage of neurons with collaterals versus without collaterals: ∼90% versus ∼10%). Chi-Square test, *P < 0.05. (H) Proposed model of AcD neuron development.

Development of neurons with axon-carrying dendrite. (A) Representative images of dissociated hippocampal primary neurons with nonAcD (top row) and AcD (bottom row) morphology at different ages. AnkG and MAP2 immunostaining indicate the axon/AIS and dendrites of a neuron, respectively. White arrowhead indicates AIS; yellow and pink arrowhead indicates AcD and stem dendrite of AcD neuron respectively; and cyan arrowhead indicates regular dendrite. Scale bar is 20 µm. (B) Schematic of AcD neuron. Black dashed line indicates the end of soma; orange dashed line indicates the beginning of an axon. Cyan solid line with double arrowheads indicates the axon distance. Magenta solid line with double arrowheads indicates the first (d1) and the second (d2) line used to calculate the averaged diameter of stem dendrite. (C) Time-lapse images of nonAcD (top row) and AcD neurons (bottom row) at different developmental stages. The precursor neurite in the displayed AcD neuron became an axon and collateral from the precursor neurite developed as an AcD. Orange arrowhead indicates the precursor neurite of AcD neuron; blue arrowhead indicates the precursor neurite of nonAcD neuron; white arrowhead indicates axon; yellow and pink arrowhead indicates AcD and stem dendrite of AcD neuron, respectively. Scale bar is 20 µm. Cell bodies with dendrites and the AIS are labeled by MAP2 and AnkG staining, respectively. Related to Videos 1 and 2. (D) Percentage of AcD neurons with a generated collateral or bifurcation of the precursor neurite. 1 culture preparation, AcD (total): n = 27 cells, collateral: n = 22 cells (∼80%), bifurcation: n = 5 cells (∼20%). (E) Time points of collateral formation at the precursor neurite in AcD neurons. One neuronal culture preparation, AcD (total): n = 27 cells, time points: 27 h (n = 3 cells), 33 h (n = 7 cells), 39 h (n = 9 cells), 45 h (n = 4 cells), 51 h (n = 4 cells). (F) Percentage of AcD neurons developing the precursor neurite into an axon or an AcD. 1 culture preparation, AcD: n = 22 cells, precursor as axon: n = 16 cells (∼70%), precursor as AcD: n = 6 cells (∼30%). (G) Percentage of nonAcD and AcD neurons that formed collaterals at the proximal region of the precursor neurite during development. One culture preparation, nonAcD: n = 25 cells (percentage of neurons with collaterals versus without collaterals: ∼32% versus ∼68%), AcD: n = 23 cells (percentage of neurons with collaterals versus without collaterals: ∼90% versus ∼10%). Chi-Square test, *P < 0.05. (H) Proposed model of AcD neuron development.

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