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. 2022 Oct 24;16:994735. doi: 10.3389/fnins.2022.994735

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Copyright © 2022 Hingorani, Viviani, Sanfilippo and Janušonis.

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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(A) Primary midbrain culture (monolayer at DIV 4), visualized with immunocytochemistry for 5-HT (red) and MAP2 (green) and imaged with high-resolution confocal microscopy. Cell nuclei are stained blue (DAPI). (A,B) Four branching regions (white 1–4) and two growth cones (yellow 1–2) and are shown. Note that the two branches in (B) differ in their morphologies. (C) An enlarged view of the branching points. The branching regions have a triangular shape, and the branches diverge at large angles (around 90°–180°). (D) An enlarged view of the growth cones. Note the similarity of the second growth cone to that in the embryonic mouse brain (Figure 9). Scale bars = 10 μm in (A,B), 5 μm in (C,D).