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. 2016 Mar 15;5:e13399. doi: 10.7554/eLife.13399

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© 2016, Lacin et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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Figure 5. — (A) R10C12 marks uniquely Msh⁺ NB2-3 (arrows) in each thoracic hemisegment but not in abdominal ganglia. Dpn, blue; Msh, red (B) A lineage clone containing NB2-3 (arrow) extends only efferent axons, which splits into two bundles after exiting the CNS via the segmental nerve (SN). (C) Immortalization of R10C12 expression in the NB2-3 progeny marks lineage 15, which is composed of motor neurons (arrowhead) and glia (arrow). (D) T3 and A1 segments from a stage 12 embryo shown. eg-lacZ (red) and Msh (green) co-staining reveal differences in NB (blue) pattern of T3 and A1 segments. Schematic representation is shown on the right. Msh marks three thoracic specific NBs (NB2-3, NB3-4, and NB5-7; white arrowheads). NB3-4, which is marked by both eg-lacZ and Msh, resides medial to NB3-3. In abdominal segments, no NB with lacZ and Msh expression is detected medial to NB3-3; note the region immediately medial to NB3-3 does not have any NB (asterisks). (E-I) R77B09^AD-R28H10^DBD used to identify the postembryonic progeny of NB2-3 and NB3-4. (E-G) Nerve cords from late stage embryos shown. NB3-4 (arrowhead) and its progeny express Eg (red) while NB2-3 (arrow) and its progeny do not. Inset shows the magnified views of NB2-3 (left) and NB3-4 (right) lineages. (F) A lineage clone via R77B09^AD-R28H10^DBD separates two NB lineages: NB3-4 lineage is marked with red. Progeny of NB3-4 (H) extend axons out of the CNS using the SN route like NB2-3 progeny (G) but do not extend them as far as NB2-3 progeny (see inset H). (I) Nerve cord from a late-stage larva shown. Msh, red; Dpn, blue. Immortalization of R77B09^AD-R28H10^DBD expression marks lineage 15 (NB2-3 progeny) and lineage 25 (NB3-4 progeny). (J) Immortalization of eg-GAL4 marks Toy expressing lineage 8 (NB3-3 progeny; red arrowheads; see also Figure 4—figure supplement 1L) and lineage 25 (white arrowheads). (K) Lineage 25 contains glial cells (arrow). (L-M) elav-MARCM clones showing lineage 25 have a small cluster of Msh expressing cells (arrowheads) and two Nkx6 expressing motor neurons (arrows). Msh expressing cells reside closely to Msh⁺ NB3-4, but do not extend any processes (arrowheads in L and M). Note glial cells in lineage 25 are not visible in MARCM clones since elav-GAL4 does not mark glial cells. White bar indicates the midline. FO, flip-out lineage clone; IM, immortalization; A.C., anterior commissure; P.C., posterior commissure; white bar, midline.

DOI: http://dx.doi.org/10.7554/eLife.13399.014

Figure 5—figure supplement 1. — (A) R10C12 marks uniquely Msh⁺ NB2-3 (arrows) in each thoracic hemisegment but not in abdominal ganglia. Dpn, blue; Msh, red (B) A lineage clone containing NB2-3 (arrow) extends only efferent axons, which splits into two bundles after exiting the CNS via the segmental nerve (SN). (C) Immortalization of R10C12 expression in the NB2-3 progeny marks lineage 15, which is composed of motor neurons (arrowhead) and glia (arrow). (D) T3 and A1 segments from a stage 12 embryo shown. eg-lacZ (red) and Msh (green) co-staining reveal differences in NB (blue) pattern of T3 and A1 segments. Schematic representation is shown on the right. Msh marks three thoracic specific NBs (NB2-3, NB3-4, and NB5-7; white arrowheads). NB3-4, which is marked by both eg-lacZ and Msh, resides medial to NB3-3. In abdominal segments, no NB with lacZ and Msh expression is detected medial to NB3-3; note the region immediately medial to NB3-3 does not have any NB (asterisks). (E-I) R77B09^AD-R28H10^DBD used to identify the postembryonic progeny of NB2-3 and NB3-4. (E-G) Nerve cords from late stage embryos shown. NB3-4 (arrowhead) and its progeny express Eg (red) while NB2-3 (arrow) and its progeny do not. Inset shows the magnified views of NB2-3 (left) and NB3-4 (right) lineages. (F) A lineage clone via R77B09^AD-R28H10^DBD separates two NB lineages: NB3-4 lineage is marked with red. Progeny of NB3-4 (H) extend axons out of the CNS using the SN route like NB2-3 progeny (G) but do not extend them as far as NB2-3 progeny (see inset H). (I) Nerve cord from a late-stage larva shown. Msh, red; Dpn, blue. Immortalization of R77B09^AD-R28H10^DBD expression marks lineage 15 (NB2-3 progeny) and lineage 25 (NB3-4 progeny). (J) Immortalization of eg-GAL4 marks Toy expressing lineage 8 (NB3-3 progeny; red arrowheads; see also Figure 4—figure supplement 1L) and lineage 25 (white arrowheads). (K) Lineage 25 contains glial cells (arrow). (L-M) elav-MARCM clones showing lineage 25 have a small cluster of Msh expressing cells (arrowheads) and two Nkx6 expressing motor neurons (arrows). Msh expressing cells reside closely to Msh⁺ NB3-4, but do not extend any processes (arrowheads in L and M). Note glial cells in lineage 25 are not visible in MARCM clones since elav-GAL4 does not mark glial cells. White bar indicates the midline. FO, flip-out lineage clone; IM, immortalization; A.C., anterior commissure; P.C., posterior commissure; white bar, midline.

DOI: http://dx.doi.org/10.7554/eLife.13399.014