Control of Dendritic Field Formation in Drosophila

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Control of Dendritic Field Formation in Drosophila Fen-Biao Gao, Minoree Kohwi, Jay E. Brenman, Lily Yeh Jan, Yuh Nung Jan  Neuron  Volume 28, Issue 1, Pages 91-101 (October 2000) DOI: 10.1016/S0896-6273(00)00088-X

Figure 1 Drosophila PNS md Neurons Have Their Own Dendritic Fields as Revealed by Laser Ablation Experiments (A) Schematic representation of dorsal clusters of embryonic PNS neurons. There are six md neurons (ovals), four es neurons (circles), one bd neuron (diamond), and one tracheal innervating neuron (square). GFP is expressed (green) in all but the four es neurons (gray) in the dorsal cluster. (B) A confocal image of the dorsal cluster neurons taken from a living embryo. (C) Tracing of dendritic branching pattern of the md neuron indicated by the arrow in (A). (D) Several neurons in this dorsal cluster were ablated using a laser beam at 16–17 hr AEL, and the image was taken 5–6 hr later. The yellow arrow indicates the dendrites of surviving neurons still extending toward the posterior segment boundary. (E) Tracing of the same md neuron as in (C). (F) The remaining neurons in the ablated cluster still extend their dendrites toward the anterior segment boundary as they do in a normal segment. The asterisk indicates the corps of ablated neurons. Neuron 2000 28, 91-101DOI: (10.1016/S0896-6273(00)00088-X)

Figure 2 Dendritic Fields of md Neurons in Proneural Gene Mutant Embryos or Larvae (A and B) Lateral cluster md neurons in two adjacent abdominal segments in atonal mutant first instar larva. The md neuron in the lateral cluster of the A3 segment (A) labeled by the plus sign is absent in the adjacent segment (B). The md neuron indicated by the asterisk does not extend dendrites to cover the area (indicated by the square) normally covered by dendrites extending from the md neuron that is denoted by the plus sign. (C) Schematic representation of all the PNS neurons except bd neurons in a dorsal cluster. (D and E) A md neuron in the dorsal cluster of an ac sc mutant embryo. The dendrites of this md neuron do not cover the whole segment, even though other md neurons are absent in the same hemisegment. The yellow lines indicate the dorsal midline and the segment boundary. Neuron 2000 28, 91-101DOI: (10.1016/S0896-6273(00)00088-X)

Figure 3 Dendritic Fields of Dorsal Cluster md Neurons in the Third Instar Larval Stage (A) The dorsal cluster of md neurons lies adjacent to the segment in which a few md neurons were ablated during embryogenesis. The white arrowhead indicates the cell body of the md neuron that sends its dendrites to cover the area indicated by the white square. The red plus signs indicate the cell bodies of the md neurons sending out dendrites to cover the area indicated by the red square. (B) The white plus signs indicate the positions of ablated md neurons that are homologous to the neurons indicated by the red plus signs in the adjacent segment (A). (C and D) The red lines in (C) and (D) indicate the dorsal midline. The red arrow in (C) indicates an example where the dendritic terminal branches of homologous neurons from the two contralateral hemisegments meet. Neuron 2000 28, 91-101DOI: (10.1016/S0896-6273(00)00088-X)

Figure 4 Time Lapse Analysis of Dorsal Dendrite Extension in Wild-Type and fmi72 Mutant Embryos Dendrite development beginning at 21 hr AEL was recorded at 25°C using confocal microscopy. Pictures were taken at 10 min intervals. The white arrows in the wild-type embryo panels point to the fine processes at the tips of dorsal dendrites. The white arrowhead in fmi72 mutant embryo panels indicates the dorsal dendrite that was already overextended at the beginning of the recording. The white arrows indicate the dorsal dendrites that were extending during the recording. Scale bar: 20 μm. Neuron 2000 28, 91-101DOI: (10.1016/S0896-6273(00)00088-X)

Figure 5 The Absence of Competition between Dendrites of Homologous md Neurons in fmi72 Mutant Larvae The yellow lines in (A)–(D) and (F) indicate the dorsal midline. (A) A newly hatched wild-type larva. (B) A newly hatched fmi72 mutant larva. (C) A fmi72 mutant larva 10–12 hr after hatching. The asterisks indicate the ablated dorsal clusters of md neurons. The ablation was done during embryogenesis. (D) A wild-type larva 10–12 hr after hatching. (E) Tracing of dendrites near the dorsal midline in (D). (F) A fmi72 mutant larva 1–2 hr after hatching. (G) Tracing of dendrites near the dorsal midline in (F). Neuron 2000 28, 91-101DOI: (10.1016/S0896-6273(00)00088-X)

Figure 6 Flamingo Expression on Dendrites (A) Simultaneous labeling of md neuron dendrites and adjacent epithelial cells by GFP. (B) Expression of Flamingo on dorsal cluster es, md, and bd neurons. The arrows indicate dendrites of md and bd neurons separately, and the arrowhead indicates the axons. (C) A face-down view of Flamingo expression on epithelial cells. The bracket indicates the dorsal cluster neurons. (D) Flamingo is localized to adherens junctions. Neuron 2000 28, 91-101DOI: (10.1016/S0896-6273(00)00088-X)

Figure 7 Cell-Autonomous Function of Flamingo in Dendrite Outgrowth (A) A wild-type embryo. (B) Dendritic overextension in a fmi72 mutant embryo. (C) Dendritic phenotype in a fmi72 mutant embryo expressing UAS-Flamingo. (D) Dendritic overextension phenotype in a wild-type embryo overexpressing Flamingo. (E) Dendritic branching patterns of dorsal md neurons in a wild-type first instar larva. (F) Dendritic overextension phenotype in a first instar larva overexpressing Flamingo. Gal4 109(2) 80 was used in all experiments except (C), in which both Gal4 109(2) 80 and Gal4 1407 were used. Neuron 2000 28, 91-101DOI: (10.1016/S0896-6273(00)00088-X)