Ariane Zierau’s research while affiliated with University of Münster and other places

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Publications (4)


Figure 1. Dscam mutant axons converge class-specifically in ectopic spots. (A,B) Wild type ORN47a axons (red, anti-CD2) grow to their specific target and converge into one glomerulus (green, sytGFP) (A) whereas Dscam mutant axons converge into an ectopic glomerular-like structure (B). (C-J) Double labelling of different ORN classes show ORN class-specific axon sorting. Dscam mutant ORN47a axons converge in the neighbourhood of ORN22a glomerulus but the axons never intermingle (D). Neighbouring projecting ORN classes 47b and 88a show distinct boundaries in wild type (E,E') as well as in Dscam mutants (F,F'). (G,H) Dscam mutant axons coming from the maxillary palps converge into distinct ectopic spots at the border to the AL or at the suboesophagial ganglion (SOG) (asterisks in H"). Ectopic projecting ORN46a axons converge into one distinct spot in the neighborhood of other ectopic spots (H') and they never intermingle with other ORN classes e.g. ORN71a (J). SOG: suboesophagial ganglion. Green: (A-J) sytGFP; blue: (A,B) N-Cad, (C-J) Toto3; red: (A-J) ratCD2. Scale bar: 25 µm. Genotype: (A) eyflp UAS-CD2; FRT42 47a::sytGFP/FRT42 PCNA; 47a-Gal4 UAS-CD2. (B) eyflp UAS-CD2; FRT42 Dscam 47a::sytGFP/FRT42 PCNA; 47a-Gal4 UAS-CD2. (C) eyflp UAS-CD2; FRT42 47a::sytGFP/FRT42 PCNA; 22a-Gal4 UAS-CD2. (D) eyflp UAS-CD2; FRT42 Dscam 47a::sytGFP/FRT42 PCNA; 22a-Gal4 UAS-CD2. (E) eyflp UAS-CD2; FRT42 47b::sytGFP/FRT42 PCNA; 88a-Gal4 UAS-CD2. (F) eyflp UAS-CD2; FRT42 Dscam 47b::sytGFP/FRT42 PCNA; 88a-Gal4 UAS-CD2. (G) eyflp; FRT42/FRT42 PCNA; 46a::sytGFP/MT14-Gal4 UAS-CD2. (H) eyflp; FRT42 Dscam/FRT42 PCNA; 46a::sytGFP/ MT14-Gal4 UAS-CD2 (I) eyflp UAS-CD2; FRT42/FRT42 PCNA; 46a::sytGFP 71a-Gal4 UAS-CD2. (J) eyflp UAS-CD2; FRT42 Dscam/FRT42 PCNA; 46a::sytGFP 71a-Gal4 UAS-CD2.
Figure 2. Pre-and postsynaptic recognition is Dscam independent. (A) Schematic showing ORN/PN/LN matching in olfactory lobes of Drosophila in wild type and Dscam mutants. (B-E) ORN-PN matching identities remain in Dscam mutants. Mz-19 positive PN dendrites connect to axons of ORN class 88a (D) and not to 47b (B) in wild type. In Dscam mutant, the dendrites follow the misprojecting 88a axons (E) but avoid ectopic 47b axons (C). (F-J) GH146 expressing dendrites do not innervate ectopic Dscam mutant ORN 47a spots, when they are far away from the wild type glomerulus (box in H, compared to F). Ectopic spots of ORN46a axons outside of the AL are not innervated by GH146-positive dendrites (G,J) even if the glomerulus of the ORN46a class is innervated by GH146-positive dendrites. (K-N) Ectopic Dscam mutant spots are innervated from C753-positive LNs in the AL in case of ORN21a (L) as well as outside the AL in case of ORN46a (N). Green: sytGFP, red: ratCD2, blue: Toto3. Scale bar: 25 µm. Genotype: (B) eyflp UAS-CD2; FRT42/FRT42 PCNA; 47b::sytGFP Mz19-Gal4 UAS-CD2. (C) eyflp UAS-CD2; FRT42 Dscam/FRT42 PCNA; 47b::sytGFP Mz19-Gal4 UAS-CD2. (D) eyflp UAS-CD2; FRT42/FRT42 PCNA; 88a::sytGFP Mz19-Gal4 UAS-CD2. (E) eyflp UAS-CD2; FRT42 Dscam/FRT42 PCNA; 88a::sytGFP Mz19-Gal4 UAS-CD2. (F-G) eyflp UAS-CD2; FRT42 OR::sytGFP/ FRT42 PCNA; GH146-Gal4 UAS-CD2. (H-J) eyflp UAS-CD2; FRT42 Dscam OR::sytGFP/FRT42 PCNA; GH146-Gal4 UAS-CD2. (K,M) eyflp UAS-CD2; FRT42 OR::sytGFP/FRT42 PCNA; C753-Gal4 UAS-CD2. (L,N) eyflp UAS-CD2; FRT42 Dscam OR::sytGFP/FRT42 PCNA; C753-Gal4 UAS-CD2.
Figure 3. Formation of ectopic glomeruli requires inter-axonal recognition. (A,B) Multicolour axon labeling using flybow shows targeting of individual mutant ORN 47a axons in WT (A-A") and Dscam (B-B") mutants. The ectopic glomeruli in Dscam mutants always consisted of multiple axons (marked with arrow heads in B' and B"). (C,D) Visualizing targeting of single/few cell OR47a clones in WT (C) and Dscam mutants (D) showed similar phenotype with OR47a axons not mis-targetting in Dscam mutants. The number of ORNs was confirmed by visualizing cell bodies in the left and right antenna. Blue: Ncad (A,B), Red: Ncad (C,D). Scale bar: 25 µm. Genotype: (A) eyflp; FRT42/FRT42 PCNA; UAS-FB1.1, Or47a-Gal4/hs-mflp5 (B) eyflp; FRT42 dscam 21 / FRT42 PCNA; UAS-FB1.1, Or47a-Gal4/hs-mflp5 (C) eyflp; FRT42/FRT42 PCNA; UAS-FB1.1, Or47a-Gal4/hsmflp5 (D) hsflp; FRT42 dscam 21 /FRT42 Gal80; Or47a-Gal4, UAS-mCD8::GFP.
Figure 4. Dscam acts cell-autonomously. (A-H) Ectopic spots inside (A, arrow heads) as well as outside (B, arrow head) the AL are innervated only from homozygous Dscam mutant axons, whereas homozygous wild type axons always reach their wild type glomerulus (C,D, dotted circle). Presence of Dscam mutant axons is indicated by the presence of ectopic spots outside the AL, near the V glomerulus (asterisks). Over-expression of one single Dscam-isoform in wild type axons leads to disrupted glomerular pattern and axon termini are spread over a large area (E,F). Over-expression in Dscam mutant axons shows the same phenotype than the over-expression in wild type (G,H). Only the early stopping phenotype outside the AL of maxillary ORN axons can be rescued by over-expression of a single Dscam-isoform (H). (I-T) Broad over-expression of a single Dscam-isoform in ORNs show an AL wide distribution of axon termini (green in J) and a complete loss of the glomerular structure (red in J). Dscam over-expression affects only ORN axons, in which it is expressed. On over-expression in the con-positive ORNs, the structure of the glomerulus innervated from ORN class 47b is totally disrupted (L) whereas the ORN class 47a, which project neighboring to the con-positive domain, is unaffected (N). Over-expression of a single Dscam isoform in single ORN axons also reveal a misprojecting phenotype (P,R,T). Green: sytGFP, blue: Toto3, red: (A-J,O,P) N-cad, (K-N,Q-T) ratCD2. Scale bar: 25 µm. Genotype: (A,B) eyflp; FRT42 Dscam/FRT42 Gal80; OR-Gal4 UAS-sytGFP. (C,D) eyflp; FRT42 Dscam Gal80/ FRT42; OR-Gal4 UAS-sytGFP. (E,F) eyflp elav-Gal4; FRT42 OR::sytGFP/FRT42 Gal80; UAS-Dscam 17.2-7 . (G,H)
Figure 5. Over-expression of single Dscam isoform prevents targeting in a single neuron. (A,D) Single labelled wild type ORN axons from the antenna (A) and the maxillary palp (D) showing branching inside one glomerulus (A' ,D') and the contralateral branch. (B,C,E) Over-expression of a single Dscam isoform leads to defective branching inside the WT glomerulus (B' ,E') as well as Dscam mutant (C'). White: mCD8::GFP, Red: N-cad. Scale bar: 25 µm. Genotype: (A,D) hsflp, elav-Gal4 UAS-mCD8::GFP; FRT42 Gal80/FRT42. (B,E) hsflp, elav-Gal4 UAS-mCD8::GFP; FRT42 Gal80/FRT42; UAS-Dscam 17.2-7 . (C) hsflp, elav-Gal4 UAS-mCD8::GFP; FRT42 Gal80/FRT42 Dscam; UAS-Dscam 17.2-7 .

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Inter-axonal recognition organizes Drosophila olfactory map formation
  • Article
  • Full-text available

August 2019

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210 Reads

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7 Citations

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Ariane Zierau

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Marc Lattemann

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[...]

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Olfactory systems across the animal kingdom show astonishing similarities in their morphological and functional organization. In mouse and Drosophila, olfactory sensory neurons are characterized by the selective expression of a single odorant receptor (OR) type and by the OR class-specific connection in the olfactory brain center. Monospecific OR expression in mouse provides each sensory neuron with a unique recognition identity underlying class-specific axon sorting into synaptic glomeruli. Here we show that in Drosophila, although OR genes are not involved in sensory neuron connectivity, afferent sorting via OR class-specific recognition defines a central mechanism of odortopic map formation. Sensory neurons mutant for the Ig-domain receptor Dscam converge into ectopic glomeruli with single OR class identity independent of their target cells. Mosaic analysis showed that Dscam prevents premature recognition among sensory axons of the same OR class. Single Dscam isoform expression in projecting axons revealed the importance of Dscam diversity for spatially restricted glomerular convergence. These data support a model in which the precise temporal-spatial regulation of Dscam activity controls class-specific axon sorting thereby indicating convergent evolution of olfactory map formation via self-patterning of sensory neurons.

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Transcriptional Regulation of Peripheral Glial Cell Differentiation in the Embryonic Nervous System of Drosophila

September 2011

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26 Reads

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12 Citations

Glia

The Drosophila nervous system is ideally suited to study glial cell development and function, because it harbors only relatively few glial cells, and nervous system development is very well conserved during evolution. In the past, enhancer trap studies provided tools allowing to study glial cells with a single-cell resolution and, moreover, disclosed a surprising molecular heterogeneity among the different glial cells. The peripheral nervous system in the embryo comprises only 12 glial cells in one hemisegment and thus offers a unique opportunity to decipher the mechanisms directing glial development. Here, we focus on transcriptional regulators that have been reported to function during gliogenesis. To uncover additional regulators, we have conducted a genetic screen and report the identification of two additional transcriptional regulators involved in the control of peripheral glial migration: nejire and tango.


Notch and Numb are required for normal migration of peripheral glia in Drosophila

February 2007

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85 Reads

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46 Citations

Developmental Biology

A prominent feature of glial cells is their ability to migrate along axons to finally wrap and insulate them. In the embryonic Drosophila PNS, most glial cells are born in the CNS and have to migrate to reach their final destinations. To understand how migration of the peripheral glia is regulated, we have conducted a genetic screen looking for mutants that disrupt the normal glial pattern. Here we present an analysis of two of these mutants: Notch and numb. Complete loss of Notch function leads to an increase in the number of glial cells. Embryos hemizygous for the weak Notch(B-8X) allele display an irregular migration phenotype and mutant glial cells show an increased formation of filopodia-like structures. A similar phenotype occurs in embryos carrying the Notch(ts1) allele when shifted to the restrictive temperature during the glial cell migration phase, suggesting that Notch must be activated during glial migration. This is corroborated by the fact that cell-specific reduction of Notch activity in glial cells by directed numb expression also results in similar migration phenotypes. Since the glial migration phenotypes of Notch and numb mutants resemble each other, our data support a model where the precise temporal and quantitative regulation of Numb and Notch activity is not only required during fate decisions but also later during glial differentiation and migration.


Semaphorin-1a Controls Receptor Neuron-Specific Axonal Convergence in the Primary Olfactory Center of Drosophila

February 2007

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34 Reads

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55 Citations

Neuron

In the olfactory system of Drosophila, 50 functional classes of sensory receptor neurons (ORNs) project in a highly organized fashion into the CNS, where they sort out from one another and converge into distinct synaptic glomeruli. We identified the transmembrane molecule Semaphorin-1a (Sema-1a) as an essential component to ensure glomerulus-specific axon segregation. Removal of sema-1a in ORNs does not affect the pathfinding toward their target area but disrupts local axonal convergence into a single glomerulus, resulting in two distinct targeting phenotypes: axons either intermingle with adjacent ORN classes or segregate according to their odorant receptor identity into ectopic sites. Differential Sema-1a expression can be detected among neighboring glomeruli, and mosaic analyses show that sema-1a functions nonautonomously in ORN axon sorting. These findings provide insights into the mechanism by which afferent interactions lead to synaptic specificity in the olfactory system.

Citations (4)


... The litany of cell surface proteins implicated in olfactory system development, such as Semaphorins, DSCAM, Tenascins/Teneurins and Toll receptors, suggests that ORN class-specific cell surface codes are the main drivers of this topographic circuit assembly. 9,11,[13][14][15][16][17][18] Though there are several examples of cell surface molecules regulating ORN class-specific axon organization, it is still unclear when and how ORN class-specific axon convergence occurs during olfactory circuit development. 4,15,16,19 Given the importance of cell surface combinatorial codes, identifying novel cell surface proteins functionally required for olfactory circuit assembly is critical to decrypting the ''rosetta stone'' needed to translate cell surface signatures into predictable discrete cellular processes. ...

Reference:

Atypical cadherin, Fat2, regulates axon terminal organization in the developing Drosophila olfactory receptor neurons
Inter-axonal recognition organizes Drosophila olfactory map formation

... Another putative target gene of Hb is Ets98b (Fig 8B), the ortholog of which has recently been shown to induce ectopic cell migration upon misexpression during early embryonic development in the common house spider Parasteatoda tepidariorum [152]. Finally, Nejire has been shown to be involved in glia cell migration in the peripheral nervous system in Drosophila [153], and we found it as putative regulator of genes in cluster 13. It remains to be established, whether Nejire and Hb may collaborate during carpet cell development. ...

Transcriptional Regulation of Peripheral Glial Cell Differentiation in the Embryonic Nervous System of Drosophila
  • Citing Article
  • September 2011

Glia

... This experiment revealed an up-regulation of the LacZ signal following Delta knockdown in mCherry-positive nuclei in the vicinity of M4, indicating increased Mmp1 transcription ( Fig. 1 M and N). Our findings demonstrate the presence of Delta in late stages of larval life both in neurons and glia of the CNS and along motor nerve bundles (44,45), and identify Delta in SPG as a critical regulator of Mmp1 expression. ...

Notch and Numb are required for normal migration of peripheral glia in Drosophila
  • Citing Article
  • February 2007

Developmental Biology

... 8 Between 20 hAPF and 40 hAPF, the exploratory axons target a glomerulus, 9,10 interact with PNs and local interneurons (LNs), 11,12 and then begin to simultaneously repel from adjacent glomeruli while converging with class-specific axons to further accentuate proto-glomerular boundaries. 4,13 The mechanisms critical to the stabilization of these transient axonal branches are unknown, and likely involve proteins that link chemoaffinity receptor activity to cytoskeletal regulators. The litany of cell surface proteins implicated in olfactory system development, such as Semaphorins, DSCAM, Tenascins/Teneurins and Toll receptors, suggests that ORN class-specific cell surface codes are the main drivers of this topographic circuit assembly. ...

Semaphorin-1a Controls Receptor Neuron-Specific Axonal Convergence in the Primary Olfactory Center of Drosophila
  • Citing Article
  • February 2007

Neuron