Journal: PLoS Biology
Article Title: Genetic Dissection of the Function of Hindbrain Axonal Commissures
Figure Lengend Snippet: Reduced internuclear commissure in Robo3 knockout mice. (A to I) show coronal hindbrain sections at the level of the abducens nucleus, visualized by Hb9 immunostaining (in A, D, E, F, and H). (A, B, and C) illustrate the projection of abducens axons (arrowheads) across the midline (dashed line) in Robo3 +/− E13 embryos. Some GFP+ axons originate from the abducens nucleus (VI) and are immunoreactive for Robo3 (B and C). (D) The internuclear commissure (arrowhead) is also observed in P0 controls, following DiI injection at the level of the oculomotor nucleus III. (E) This commissure is almost completely absent in P0 Krox20::cre;Robo3 lox/lox mice (arrowhead). (VIIn): Genu of facial nerve. (F to I) At E15, many neurofilament+ axons cross the midline at the VI level (arrowheads) in control embryo (F and G), whereas they are rare in Krox20::cre;Robo3 lox/lox embryo (H and I). Scale bars represent 100 µm, except in (G and I), where they indicate 50 µm.
Article Snippet: Lee, Philadelphia, PA), mouse anti-Islet1 (1∶100, Developmental Studies Hybridoma Bank, University of Iowa), mouse anti-parvalbumin (1∶1,000, Swant), mouse anti-Robo3 (1∶200, R & D which recognizes the N-terminal domain of Robo3), guinea pig anti-VGLUT2 (1∶1,000, Millipore), rabbit anti-Hb9 (1∶1,000, Abcam), rabbit anti-CGRP (1∶1,000, Peninsula), rabbit anti-GFP (1∶300, Invitrogen), rabbit anti-Tag1 (1∶3,000, gift from Dr. Domna Karagogeos, University of Crete Medical School, Heraklion, Greece), rabbit anti-CaBP (1∶5,000, Swant), rabbit anti-βGal (1∶1,000, Cappel), goat anti-mouse Alcam (1∶200, R & D), goat anti-Brn3.2 (1∶200, Santa Cruz Biotechnology), goat anti-Robo3 (1∶300, R & D), goat anti-ChAT (1∶400, Millipore), and chicken anti-GFP (1∶800, Abcam).
Techniques: Knock-Out, Mouse Assay, Immunostaining, Injection